Compound astragalus membranaceus medicinal liquor, and preparation method and application thereof

Abstract

The present application belongs to the technical field of yellow rice wine preparation, and is characterized by using low-alcohol millet yellow rice wine as base wine, pulverizing radix angelicae sinensis, dried ginger, perilla fruit and dried tangerine or orange peel, immersing the medicinal materials in 65-degree liquor, oscillating and extracting at 120 r / min, and then adding yellow rice wine to the extracted liquid after concentration to homogenize; wherein, the amount of radix angelicae sinensis, dried ginger, perilla fruit and dried tangerine or orange peel is 12 g, 9 g, 9 g and 6 g respectively. The sealed extraction of the liquor can dissolve the effective components to the greatest extent, and the efficacy can be maximized. The taste and color of the product are formed completely by relying on the homologous food and ordinary food in the prescription, without any additives, which is a natural integration of the medicinal properties and food properties. The yellow rice wine as the base wine not only needs to meet the theory of traditional Chinese medicine, but also needs to consider the use, efficacy and suitable population. For those who cannot drink high-concentration liquor, we choose low-alcohol yellow rice wine as the base wine, which expands the range of the edible population.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of yellow rice wine preparation, and particularly relates to a compound astragalus membranaceus medicinal liquor and a preparation method and application thereof. BACKGROUND

[0002] Traditional Chinese medicine medicinal liquor has been used in practice for thousands of years, and can treat diseases and keep healthy. Investigation and research show that there are many problems in the market circulation of medicinal liquor. The concept of traditional Chinese medicine medicinal liquor is ambiguous, and the quality is uneven. The chemical components are complex and numerous, and lack scientific basis and clinical practice basis. As one of the main raw materials for preparing medicinal liquor, base liquor with different concentrations has different dissolution rates of effective components, and can even change the efficacy of compound astragalus medicinal liquor. Most medicinal liquor circulating in the market uses high-concentration alcohol base liquor as medicinal material solvent, and has more restrictions on the use of people. Most consumers describe the liquor taste as strong and irritating, and difficult to swallow, which limits the market promotion. At present, medicinal liquor mainly stays in the research of production process and quality control, and has not been systematically evaluated in terms of function. SUMMARY

[0003] The application provides a Suhuang liquor and a preparation method and application thereof.

[0004] The application is realized by the following technical scheme: a compound astragalus medicinal liquor, in which low-alcohol millet yellow rice wine is used as base liquor, astragalus, dried ginger, perilla fruit and dried tangerine or orange peel are crushed, 65-degree clear liquor is used to immerse the medicinal materials, the extraction liquid is concentrated and then mixed with the yellow rice wine to homogenize, wherein the astragalus is 12g, the dried ginger is 9g, the perilla fruit is 9g, and the dried tangerine or orange peel is 6g.

[0005] The method for preparing the compound astragalus medicinal liquor comprises the following steps: (1) crushing raw materials: astragalus, dried ginger, perilla fruit and dried tangerine or orange peel are crushed to 60 meshes; (2) extracting medicinal materials: the medicinal powder obtained in step (1) is immersed in 65-degree clear liquor, and the amount of liquor is 8 times that of the medicinal powder; the extraction temperature is controlled at 60 DEG C, and the extraction is performed for 0.5h; after the extraction is completed, the concentrated liquid is heated and opened to concentrate to 1 / 4 to 1 / 3 of the original volume; (3) mixing: the medicinal concentrated liquid obtained in step (2) is mixed with millet yellow rice wine, and the amount of medicinal extract in the final product is controlled to be 1 / 5 to 1 / 20 of the volume of the final product; (4) homogenizing: after mixing, the mixture is placed in a high-speed homogenizer, and homogenized at 5000rpm / min for 8min, and then stirred in a stirrer at 18000r / min for 10min to mix uniformly.

[0006] Further, the astragalus is 3-year-old Shanxi astragalus or 1-year-old Heilongjiang astragalus; and the dried tangerine or orange peel is 5-year-old Xinhui dried tangerine or orange peel.

[0007] The millet yellow wine is added in the drug concentrate, and the drug extract accounts for 1 / 5, 1 / 10 or 1 / 20 of the volume of the final product based on the final product as a calculation basis. Further, the millet yellow wine is added in the drug concentrate, and the drug extract accounts for 1 / 10 of the volume of the final product based on the final product as a calculation basis.

[0008] The application also provides application of the compound astragalus membranaceus medicinal wine or the compound astragalus membranaceus medicinal wine prepared by the method in preparation of a drug for treating pulmonary fibrosis, in preparation of a drug for treating cognitive impairment caused by diabetes, and in preparation of a drug for treating osteoporosis.

[0009] The application is caused by spleen and lung qi deficiency, blood deficiency, and yin deficiency and dry heat. In the formula, astragalus membranaceus is the monarch drug with the functions of tonifying the center, benefiting qi, producing saliva, nourishing blood, removing stagnation and relieving arthralgia, ginger is the minister drug with the functions of breaking blood, promoting qi and relieving pain, perilla fruit and orange peel are the auxiliary drugs with the functions of descending qi and dissipating phlegm, regulating qi and adjusting the center, and the wine is the ministerial drug with the functions of activating blood, dredging collaterals, assisting astragalus membranaceus, dispelling wind and eliminating cold. The whole formula has the functions of warming the center and harmonizing qi, and supporting the healthy qi and the root. The application is composed of native medicinal materials such as astragalus membranaceus and orange peel, and is commonly used in clinical treatment of lung deficiency cough and asthma, cognitive impairment, osteoporosis and the like, and has the function of warming the center and harmonizing qi. The scientific brewing, reasonable color, aroma and taste make the development of the new product specific, make the compound astragalus membranaceus medicinal wine closer to the consumer market, the flavor characteristics more perfect, and the efficacy outstanding. The compound astragalus membranaceus medicinal wine is extracted by sealed extraction of the clear liquor, which greatly dissolves the effective components and maximizes the drug efficacy. The taste and color of the product are completely formed by the homoeopathic food and ordinary food in the formula without any additives, which is the natural integration of the medicinal properties and food properties. The yellow wine as the base liquor not only needs to meet the theory of traditional Chinese medicine, but also needs to consider the use, efficacy and applicable population. For people who cannot drink high-concentration liquor, low-alcohol yellow wine is selected as the base liquor, and the scope of the edible population is expanded. BRIEF DESCRIPTION OF DRAWINGS

[0010] Figure 1 HE staining and Masson staining results of each group; in the figure: A is blank, B is model, C is compound astragalus medicinal wine and perilla seed reduced flavor formula; Figure 2 TRAP staining results of each group; in the figure: A is the blank group, B is the model group, and C is the compound astragalus medicinal wine group. DETAILED DESCRIPTION

[0011] The technical solutions in the application examples will be clearly and completely described below. The following experimental methods are conventional methods unless otherwise specified, and the instruments and equipment used are conventional laboratory instruments and equipment unless otherwise specified; the experimental materials used are purchased from conventional biochemical reagent stores unless otherwise specified.

[0012] I. Preparation process of the compound astragalus medicinal wine

[0013] 1. Experimental reagents: 65° clear fragrance type liquor, purchased from Shanxi Xinghua Liquor Group Co., Ltd., millet yellow rice wine (purchased from Shanxi Laogaojia Liquor Co., Ltd.), Fujian red koji wine (purchased from Pingtan Comprehensive Experimental Zone Huifeng Trade Co., Ltd.), Shaoxing yellow rice wine (purchased from Wuhan Yesu Food Trading Co., Ltd.), different origins and growth years of Astragalus membranaceus (Inner Mongolia, Gansu, Shanxi, Heilongjiang), dried roots of Astragalus membranaceus (purchased from medicinal material companies in the production area), Astragalus membranaceus glycoside, hesperidin, purchased from China Institute for Food and Drug Control, purity ≥98%.

[0014] 2. Preparation process of compound Astragalus membranaceus medicinal liquor: The basic preparation process of compound Astragalus membranaceus medicinal liquor is as follows: After the medicinal materials are crushed, they are tightly wrapped with filter paper and placed in a sealed tank. 65° clear fragrance type liquor is added to immerse the medicinal materials. The tank is shaken at a certain temperature on a shaking table (shaking table shaking frequency 120 r / min) to extract the medicinal materials. The extract is collected and concentrated to a certain degree. An appropriate amount of yellow rice wine is added, and the mixture is subjected to homogenization operation in a machine. After refinement, it is filled into a sealed container and sterilized to obtain the medicinal liquor.

[0015] (1) Quality control of Astragalus membranaceus in compound Astragalus membranaceus medicinal liquor: Astragalus membranaceus is mainly used in the treatment of deficiency of qi and blood, poor appetite and loose stools, middle qi collapse, chronic diarrhea and prolapse of the anus, and blood deficiency and emaciation. Modern research shows that Astragalus membranaceus has the pharmacological effects of regulating immunity, protecting cardiovascular and nervous systems, anti-tumor, and liver protection. As one of the most widely used medicinal materials in clinical practice, the quality control of Astragalus membranaceus is uneven. In order to better exert the effects of the drug, Astragalus membranaceus of the same species cultivated in different origins and growth years was selected to compare and analyze the contents of water, total ash, extract, and Astragalus membranaceus glycoside in Astragalus membranaceus from different origins.

[0016] A. Collection of Astragalus membranaceus raw medicinal material samples: Astragalus membranaceus: from July 2020 to March 2021, Astragalus membranaceus medicinal materials from Inner Mongolia, Heilongjiang, Shanxi, and Gansu were collected. Professor Zhang Xichun of Shanxi University of Chinese Medicine and Food Engineering identified them as the dried roots of Astragalus membranaceus.

[0017] B. Determination of water content, total ash content, and extract content: A certain amount of Astragalus membranaceus from each sample was accurately weighed and subjected to sample detection according to the determination method of water content, total ash content, and extract content in Astragalus membranaceus in Chinese Pharmacopoeia 2020 edition Volume I.

[0018] C. Determination of the content of Astragalus membranaceus glycoside by high performance liquid chromatography: Preparation of test solution: 4 g of Astragalus membranaceus from each sample was accurately weighed and placed in a Soxhlet extractor. 65° clear fragrance type liquor was added and soaked overnight. The extract was recovered and concentrated to dryness. The concentrated solution was dissolved in 10 ml of distilled water, extracted with 40 ml of water-saturated n-butanol four times, and the n-butanol solution was combined. The extract was concentrated to dryness by vacuum rotary evaporation. The residue was dissolved in methanol and transferred to a 10 ml volumetric flask. 1.0 ml of methanol was added and shaken. The supernatant was filtered through a 0.22 μm organic microporous filter membrane. The filtrate was obtained.

[0019] Preparation of Astragalus Glycoside Reference Solution: Weigh 0.001 g of astragalus glycoside reference substance, add methanol to a 5 ml volumetric flask, shake well. After filtration with a 0.5 μm microporous filter, transfer to a brown bottle and store in a 5℃ refrigerator.

[0020] Chromatographic conditions: Chromatographic column: octadecylsilane bonded silica gel; mobile phase: ethyl acetate-water (36:68); flow rate 0.3 ml / min; column temperature 45℃; injection volume 1.0 μL. Weigh the astragalus glycoside reference substance accurately and prepare a 0.5 mg / ml reference solution. Take the test solution and use HPLC to detect the content of astragalus glycoside in the sample. All experiments were repeated three times and the average value was taken as the detection result.

[0021] (2) Study on the quality control of pericarpium citri reticulatae in compound astragalus medicinal liquor: Pericarpium citri reticulatae is the dried mature fruit peel of the Rutaceae plant orange and its cultivated varieties. Chinese herbal medicines are generally divided into "pericarpium citri reticulatae" and "pericarpium citri reticulatae" (also known as Xinhui pericarpium citri reticulatae). This study selected pericarpium citri reticulatae varieties including Fukumatsu, Daikohou, and Xinhui pericarpium citri reticulatae. Through determination of moisture content and hesperidin content, the best pericarpium citri reticulatae raw material was selected to make some useful exploration for the quality control of pericarpium citri reticulatae decoction pieces.

[0022] A, pericarpium citri reticulatae raw medicinal material sample collection: Pericarpium citri reticulatae: from July 2020 to March 2021, collect Fukumatsu, Daikohou, and Xinhui pericarpium citri reticulatae medicinal materials, which are identified by Professor Zhang Xichun of the School of Chinese Medicine and Food Engineering, Shanxi University of Chinese Medicine as the dried mature fruit peel of the Rutaceae plant orange and its cultivated varieties.

[0023] B, determination of the moisture content of pericarpium citri reticulatae: The moisture content of pericarpium citri reticulatae was determined according to the method for determination of moisture content in Chinese Pharmacopoeia 2020 edition.

[0024] C, determination of the content of hesperidin by high performance liquid chromatography: Preparation of test solution: The pericarpium citri reticulatae sample was crushed through a No. 3 sieve, and about 0.1 g was weighed into 25 ml of 65° aromatic white wine, sealed and extracted for 6 hours. After ultrasonic extraction (power 500 W, frequency 40 kHz) for 40 min, it was cooled, the weight loss was made up with methanol, shaken well, and centrifuged at 15000 r / min for 10 min. The supernatant was taken as the test solution. 1 ml was taken into a sample bottle for use. Preparation of reference solution: Weigh the hesperidin reference substance accurately, dissolve in methanol to prepare a 0.4 mg / ml reference solution. Chromatographic conditions: octadecylsilane bonded silica gel chromatographic column; mobile phase: methanol-acetic acid-water (36:4:61); flow rate 1 ml / min; column temperature 35℃; injection volume 1.0 μL; detection wavelength 283 nm. Weigh the reference solution and test solution accurately, and use HPLC to detect the content of hesperidin in the sample. All experiments were repeated three times and the average value was taken as the detection result.

[0025] (3) Optimization of alcohol extraction process conditions for traditional Chinese medicine composition

[0026] A, the preferred test design of alcohol extraction process conditions: according to the preparation of compound Huangqi liquor, through different extraction factors, extraction level, design (3 4 ) orthogonal test, the alcohol extraction concentration (A), extraction temperature (B) and extraction time (C) three factors were investigated, and the experimental design was shown in table 1.

[0027] Table 1: alcohol extraction (3 4 ) orthogonal test factors and levels

[0028]

[0029] B, the determination of the yield of extract: take 20 ml of each test concentrate into the constant weight evaporating dish, evaporate in water bath, put it in 105℃ drying oven for 3h, cool for 30min, weigh, calculate according to the formula: ; in the formula, m 浸膏 is the extract mass (g), V 总 is the total volume of the extract (ml), V is the volume of the extract before drying (ml), and m is the total mass of the medicinal materials (g).

[0030] C, determination of total flavonoids: chromatographic conditions: Venusil MP C 18 chromatographic column (250 mmx4.6mm, 5μm); 0.2% formic acid water (A)-acetonitrile (B) gradient elution (0~20 min, 20%~40% B; 20~40 min, 40% B); column temperature is 30℃; ultraviolet detection wavelength is 230, 260nm; flow rate is 1 mL·min -1 ; respectively, 10 μL of the control solution and the test solution were precisely taken and injected for determination. Preparation of the control solution: accurately weigh 10 mg of the control rutin, dissolve and constant volume to 25 mL with anhydrous ethanol to obtain a 0.4 mg / mL rutin control solution. Preparation of the test solution: take about 0.2 g of sample powder (pass through a No. 4 sieve), accurately weigh, put it in a round bottom flask, accurately add 50 mL of methanol, weigh, heat reflux for 4h, cool, weigh again, add methanol to make up the lost mass, shake well, filter, accurately take 25 mL of the filtrate, recover the solvent to dryness, dissolve the residue with methanol, transfer to a 5 mL volumetric flask, add methanol to the mark, shake well, pass through a microporous filter membrane, and obtain the filtrate.

[0031] 3, the deployment technology research of compound Huangqi liquor

[0032] (1) Three-factor mixed level full cross experiment of yellow rice type, drug composition concentration and emulsification process: Daixian millet yellow rice, yellow rice is made of rice, millet or corn as raw material, with rich and mellow aroma, fresh and sweet taste; Fujian red koji wine, composed of red koji, glutinous rice, water as raw material, pure and mellow fermented wine aroma, no odor, sweet and delicious, mellow and sweet, delicate taste; Shaoxing yellow rice, traditional Shaoxing rice adheres to unique handmade process, often with glutinous rice, wheat and other cereals as raw materials, with rich aroma, mellow taste and delicate taste. The extracted solution was heated and concentrated to 1 / 4 to 1 / 3 of the original volume, and appropriate amount of millet yellow rice, Fujian yellow rice and Shaoxing yellow rice were added, so that the drug composition content in each 100 ml compound astragalus medicinal liquor was 1 / 5, 1 / 10 and 1 / 20 of the whole, respectively, and was put into high speed homogenizer (5000 rpm / min, 8 min) and dispersion stirrer (18000 r / min, 10 min), respectively. The emulsion stability was determined, and the types of yellow rice, drug composition concentration and emulsification process were investigated.

[0033] (2) Determination of creaming index: 10 ml of freshly prepared wine sample was poured into a measuring cylinder, sealed with sealing film to prevent evaporation, and placed at room temperature for static storage. The delamination status was observed and recorded. The emulsion delamination height was measured, and the creaming index CI was calculated according to the following formula to characterize the emulsion stability. ; In the formula: H S is the height of clear liquid layer; H T is the overall height of the sample.

[0034] (3) Sensory evaluation: Take appropriate amount of sample, and ask 15 experienced professionals to evaluate the color, organization form, flavor and taste of each compound astragalus medicinal liquor. The specific evaluation criteria are shown in Table 2. The final score of the sample is the average score after removing the highest and lowest scores, with a full score of 100.

[0035] Table 2: Sensory evaluation criteria of compound astragalus medicinal liquor

[0036]

[0037] II. Results and analysis

[0038] 1. The use of astragalus in compound astragalus medicinal liquor

[0039] A, Comparison of moisture content, total ash content and water extract content of Astragalus membranaceus from different producing areas and growth years: The 2020 edition of the People's Republic of China Pharmacopoeia stipulates that the moisture content of Astragalus membranaceus should not exceed 10.0%, the total ash content should not exceed 5.0%, and the water extract content should not be less than 17.0%. The experimental results are shown in Table 3. The results show that the moisture content, total ash content and water extract content of Astragalus membranaceus from different producing areas and growth years all meet the requirements of the Pharmacopoeia. The extract content of Astragalus membranaceus from Shanxi producing area with three years of growth is significantly different from that of two years and one year, and the extract content of two years of growth is not significantly different from that of one year. The extract content of Astragalus membranaceus from Gansu producing area with three years of growth is significantly different from that of two years and one year, and the extract content of two years and one year also has significant difference. The extract content of Astragalus membranaceus from Gansu producing area with two years of growth is not significantly different from that of Shanxi producing area with three years of growth. The extract content of Astragalus membranaceus from Heilongjiang producing area with one year of growth is significantly different from that of two years and three years. The extract content of Astragalus membranaceus from Inner Mongolia producing area with different years is not significantly different.

[0040] Table 3: Comparison of water extract content of Astragalus membranaceus from different producing areas and growth years

[0041]

[0042] Note: The same column has significant difference (P<0.05).

[0043] B, Determination results of astragaloside content: The 2020 edition of the People's Republic of China Pharmacopoeia stipulates that the content of astragaloside (C 41 H 68 O 14 ) should not be less than 0.080%. The analysis results are shown in Table 4. The astragaloside content of Astragalus membranaceus from different producing areas and growth years has obvious difference. The astragaloside content of Astragalus membranaceus from Shanxi producing area with three years of growth is significantly different from that of two years and one year, and the content of two years of growth is also significantly different from that of one year. Compared with Astragalus membranaceus from Inner Mongolia producing area, the astragaloside content of Astragalus membranaceus from Shanxi producing area with three years of growth is not significantly different from that of Inner Mongolia producing area with two years and three years, and is significantly different from that of one year. Compared with Astragalus membranaceus from Gansu producing area, the astragaloside content of Astragalus membranaceus from Shanxi producing area with three years of growth is not significantly different from that of Gansu producing area with three years, is significantly different from that of Gansu producing area with one year and two years, and the astragaloside content of Astragalus membranaceus from Heilongjiang producing area with different years is not significantly different. In summary, the highest astragaloside content is 3 years of Shanxi Astragalus membranaceus, and the lowest is 1 year of Heilongjiang Astragalus membranaceus. Therefore, the 3 years of Shanxi Astragalus membranaceus is the best raw material.

[0044] Table 4: Comparison of astragaloside content of Astragalus membranaceus from different producing areas and growth years

[0045]

[0046] Note: The same column with different letters have significant difference (P<0.05).

[0047] C, Formulation of Astragalus membranaceus timber standards: Astragalus membranaceus from Shanxi, Inner Mongolia, Gansu, Heilongjiang and different growth years were used as experimental objects, and their water content, ash content, water extract and astragaloside content were measured to analyze the content differences of astragaloside among different producing areas and different years. The results showed that the water content, ash content and water extract content of Astragalus membranaceus all met the requirements of the Pharmacopoeia, and the water extract content of different producing areas and different growth years had different degrees of difference. The water extract content of different growth years in Gansu producing area changed obviously, which objectively reflected that the extract of Astragalus membranaceus in Gansu producing area was unstable with the change of growth years. The content difference of Astragalus membranaceus in Shanxi, Inner Mongolia and Heilongjiang producing areas was not obvious, and it was considered that the reference value of the quality control of the original medicinal material was not high, and the astragaloside content was mainly considered. The determination results of astragaloside showed that the astragaloside content of Astragalus membranaceus in Shanxi producing area with three years of growth was the highest, and the quality of Astragalus membranaceus in Inner Mongolia with two years and three years of growth was not significantly different from that in Shanxi with three years of growth. For Astragalus membranaceus with the same growth years and different producing areas, the price of Astragalus membranaceus in Inner Mongolia and Gansu producing areas was 1.8 times higher than that in Shanxi producing area, and the price increased with the increase of years. Therefore, the three-year-old Astragalus membranaceus in Shanxi producing area was finally established as the basic standard of traditional Chinese medicine prescription raw material, and if other producing areas of Astragalus membranaceus were selected, in addition to meeting the basic standard requirements of the Pharmacopoeia and the price of raw material collection being affordable, the content of astragaloside should not be less than 90% of that of three-year-old Astragalus membranaceus in Shanxi.

[0048] 2, Use of pericarpium citri reticulatae in compound astragalus membranaceus medicinal liquor

[0049] A, Determination of water content in pericarpium citri reticulatae of different varieties and storage years: The Pharmacopoeia of the People's Republic of China stipulates that the water content of pericarpium citri reticulatae should not exceed 13.0%. The experimental results showed that the water content of pericarpium citri reticulatae of different varieties and growth years met the requirements of the Pharmacopoeia.

[0050] B, Detection of hesperidin content in pericarpium citri reticulatae of different varieties and storage years: There were differences in the content of effective component hesperidin in pericarpium citri reticulatae of different varieties and growth years. As shown in Table 5, the content of hesperidin in Xinhui pericarpium citri reticulatae of five years was the highest, which was significantly different from that of other varieties and storage years, and there was no significant difference in the content of hesperidin between Xinhui pericarpium citri reticulatae of three years and Dahongpao variety of five years, and the content of hesperidin in Fugu variety and Dahongpao variety was the lowest, and there was no significant difference in the content of hesperidin between Fugu variety of one year and three years. In summary, the content of hesperidin in Xinhui pericarpium citri reticulatae of five years was the highest, which was similar to the results of other researchers on the content determination of pericarpium citri reticulatae. Therefore, Xinhui pericarpium citri reticulatae with five years of storage was the best raw material.

[0051] Table 5: Comparison of hesperidin content in pericarpium citri reticulatae of different varieties and storage years

[0052]

[0053] Note: The same column with different letters have significant difference (P<0.05).

[0054] C, the development of the standard of the use of citrus peel: hesperidin as the effective component of citrus peel, belongs to flavonoids, pharmacopoeia stipulates that the minimum content shall not be less than 2.5%, that is, the content of hesperidin in each gram of citrus peel is higher than 25 mg. Because the flavonoids contained in citrus peel widely exist in citrus plants, and the flavonoids contained in the citrus peel on the market are significantly different. In order to ensure the safety and effectiveness of clinical medicine and the taste and flavor of the final product, fukumatsu, daren, and xinhui citrus peel and citrus peel with different storage time were selected as experimental objects, the moisture and main active component hesperidin were determined, and the content difference between different varieties and different storage time was analyzed. The results showed that the moisture content of citrus peel met the pharmacopoeia requirements and was different between different varieties and storage time. The determination results of hesperidin content showed that the content of xinhui citrus peel with five years of storage was the highest, which proved the conclusion of "long storage is good". Through market price investigation, it was found that the price of xinhui citrus peel was 0.9 times higher than that of ordinary citrus peel, and the price of citrus peel with the same storage time and different varieties was higher. In summary, xinhui citrus peel with five years of growth was finally established as the basic standard of traditional Chinese medicine prescription raw material. If other citrus peels are selected, in addition to meeting the basic standard requirements of pharmacopoeia and the price of raw material collection can be borne, the content of hesperidin should not be less than 90% of xinhui citrus peel with five years of growth.

[0055] 3, the optimization results of extraction process conditions

[0056] A, the best process conditions of the extraction yield of compound astragalus wine: take each test concentrate liquid to concentrate and dry, and the extraction yield results are shown in table 6, and the variance analysis of orthogonal experiment results is shown in table 7.

[0057] Table 6: the experimental results of the extraction yield of compound astragalus wine

[0058]

[0059] Table 7: variance analysis of orthogonal experiment results of extraction yield

[0060]

[0061] The order of the influence of each factor on the extraction rate of the extract was ethanol concentration (A) > extraction temperature (B) > extraction time (C) by mean calculation and range analysis. Ethanol concentration had a very significant influence on the extraction rate (P<0.01), extraction temperature had a significant influence on the extraction rate (P<0.05), and extraction time had no significant influence on the extraction rate (P>0.05). Further multiple comparisons of the influence of different ethanol concentrations on the extraction rate of the compound astragalus medicinal wine showed that there was a significant difference between 55% ethanol concentration and 60% and 65% ethanol concentration (P<0.01), and the extraction rate of 55% ethanol concentration was lower than that of 60% and 65% ethanol concentration; there was no significant difference between 60% and 65% ethanol concentration. Considering resource conservation and cost, the optimal ethanol concentration was selected as 60%. Further multiple comparisons of the influence of different extraction temperatures on the extraction rate showed that there was a significant difference between 25°C extraction temperature and 60°C and 90°C extraction temperature (P<0.05), and the extraction rate of 25°C extraction temperature was lower than that of 60°C and 90°C extraction temperature; there was no significant difference between 60°C and 90°C extraction temperature. Considering cost, the optimal extraction temperature was selected as 60°C. In summary, the optimal combination of factor levels was 60% ethanol concentration, 60°C extraction temperature, and 0.5h extraction time.

[0062] B, the optimal process conditions for extracting total flavonoids from compound astragalus medicinal wine: ethanol concentration, extraction temperature, and extraction time (3 4 ) orthogonal table was used for process condition optimization experiment, and the results are shown in Table 8. The variance analysis of the total flavonoid extraction rate is shown in Table 9.

[0063] Table 8: Results of orthogonal experiment for extracting total flavonoids from compound astragalus medicinal wine

[0064]

[0065] Table 9: Variance analysis of the results of orthogonal experiment for extracting total flavonoids

[0066]

[0067] The order of the influence of each factor on the extraction rate of total flavonoids was ethanol concentration (A) > extraction time (C) > extraction temperature (B) by mean calculation and range analysis. The results of variance analysis showed that the influence of ethanol concentration on the extraction rate of total flavonoids was extremely significant (P<0.05), and the influences of extraction time and extraction temperature on the extraction rate of total flavonoids were not significant (P>0.05). Multiple comparisons of the influence of different ethanol concentrations on the extraction rate of total flavonoids of compound Huangqi medicinal liquor showed that there was a significant difference between 55% ethanol concentration and 60% and 65% ethanol concentrations (P<0.05), and the extraction rate of total flavonoids at 55% ethanol concentration was lower than that at 60% and 65% ethanol concentrations; there was no significant difference between 60% and 65% ethanol concentrations; considering resource conservation and cost, the optimal ethanol concentration was 60%. In summary, the optimal production process of compound Huangqi medicinal liquor was as follows: ethanol concentration 60%, extraction temperature 60℃, and extraction time 0.5 h.

[0068] C. Optimal process and its verification: Based on the comprehensive analysis of cost resources and experimental data, the required process conditions were selected for experiments, the extraction rate of extract and the extraction rate of total flavonoids were determined, and three parallel experiments were conducted. The results are shown in Table 10. The results of orthogonal experiment are similar, indicating the reliability of orthogonal experiment. Therefore, the optimal alcohol extraction process is as follows: the raw medicine is extracted with ethanol with a concentration of 60%, the extraction temperature is 60℃, and the extraction time is 0.5 h.

[0069] Table 10: Verification experiment results of alcohol extraction process

[0070]

[0071] 4. Influence of different blending techniques on the creaming index of compound Huangqi medicinal liquor emulsion

[0072] Influence of drug composition concentration, emulsification process and yellow rice type on creaming index: Creaming index is the most intuitive method to characterize the stability of compound Huangqi medicinal liquor (referred to as HQJ). The smaller the creaming index, the better the stability of the emulsion. The creaming index of compound Huangqi medicinal liquor emulsion changes with the change of blending concentration and emulsification process. Different creaming phenomena occur in all treatment groups after 7 days, and different creaming indexes are shown under the same storage time, and the creaming index increases to different degrees with the increase of standing time. The results show that there is no significant difference between the influences of different types of yellow rice on the creaming index, and there is no significant difference between the emulsification processes of dispersed stirring emulsification and high-speed homogenization emulsification. There is a significant difference between the creaming index results of 1 / 5 blending concentration and 1 / 10 and 1 / 20 blending concentrations, and the creaming index is smaller when the blending concentration is 1 / 10.

[0073] A, Effect of different types of yellow rice wine on the creaming index of compound Astragalus membranaceus wine: The creaming index of compound Astragalus membranaceus wine prepared from three types of yellow rice wine increased with the increase of standing time, and there was no significant difference among the three. Considering the taste and color, etc., the sweet and fresh millet yellow rice wine was selected. The results are shown in Table 11.

[0074] Table 11: Effect of different types of yellow rice wine on the creaming index ; Note: different letters have significant difference (P<0.05).

[0075]

[0076] B, Effect of different drug combination concentrations on the creaming index of compound Astragalus membranaceus wine: There was a significant difference in the creaming index between 1 / 5 and 1 / 10, 1 / 20. The creaming index of 1 / 5 and 1 / 20 increased significantly with the increase of standing time, and the stability of the prepared compound Astragalus membranaceus wine was poor. The increase of creaming index of 1 / 10 was the smallest with the increase of standing time, and it was relatively stable. Therefore, 1 / 10 was selected as the best concentration. The results are shown in Table 12.

[0077] Table 12: Effect of drug combination concentration on the creaming index

[0078]

[0079] Note: different letters in the same column have significant difference (P<0.05).

[0080] C, Effect of different emulsification processes on the creaming index of compound Astragalus membranaceus wine: With the increase of standing time, there was no significant difference in the effect of different emulsification processes on the creaming index. There was no significant difference between the compound Astragalus membranaceus wine prepared by high-speed homogenization and stirring emulsification. Considering the tissue state and color, etc., high-speed homogenization process was selected. The results are shown in Table 13.

[0081] Table 13: Effect of different emulsification processes on the creaming index

[0082]

[0083] Note: different letters in the same column have significant difference (P<0.05).

[0084] 5、The influence of yellow rice wine type, drug composition concentration and emulsification process on the sensory performance of compound Huangqi medicinal wine: the higher the sensory performance score, the better the corresponding condition. The analysis results show that there are significant differences between different types of yellow rice wine, different drug composition concentrations and different emulsification processes, and the overall score of yellow rice wine type is relatively high. It is speculated that the reason may be that the combination of traditional Chinese medicine and yellow rice wine leads to different results. The drug of dried tangerine or orange peel has a strong traditional Chinese medicine flavor. If the concentration is too high, the wine will have too strong a traditional Chinese medicine flavor, and if it is too low, it will lack the combination flavor of yellow rice wine and traditional Chinese medicine. From the analysis of the emulsification process, the overall score of the emulsification process of the high-speed homogenizer is higher than that of the dispersion mixer. In summary, the highest score of each item is millet yellow rice wine, the ratio is 1 / 10, and the emulsification process is adjusted by the high-speed homogenizer. The resulting health wine is yellow with luster and no suspended matter, has pure aroma with slight traditional Chinese medicine aroma, has mellow taste and endless aftertaste.

[0085] A, The influence of yellow rice wine type on the sensory performance of compound Huangqi medicinal wine: there is a significant difference between the total scores of the three types of yellow rice wine. The color, texture, smell and taste scores of millet yellow rice wine are better, and the overall score is higher than that of Fujian yellow rice wine and Shaoxing yellow rice wine. Therefore, millet yellow rice wine is selected. The results are shown in Table 14.

[0086] Table 14: Influence of different yellow rice wine types on the sensory performance of compound Huangqi medicinal wine

[0087]

[0088] Note: Different letters in the same column have significant difference (P<0.05).

[0089] B, The influence of drug composition concentration on the sensory performance of compound Huangqi medicinal wine: there is a significant difference between the ratio of 1 / 5, 1 / 10 and 1 / 20. The color, texture, smell and taste scores of the ratio of 1 / 10 are higher than those of the ratio of 1 / 5 and 1 / 20. Therefore, the ratio of 1 / 10 is selected. The results are shown in Table 15.

[0090] Table 15: Influence of drug composition and concentration on the sensory performance of compound Huangqi medicinal wine

[0091]

[0092] Note: Different letters in the same column have significant difference (P<0.05).

[0093] C. Effect of emulsification process on sensory properties of compound Huangqi liquor: There was significant difference between different emulsification processes. The scores of color, texture, odor and taste of the product obtained by high-speed homogenization were higher than those obtained by dispersion stirring emulsification. Therefore, high-speed homogenization was selected as the emulsification process. The results are shown in Table 16.

[0094] Table 16: Effect of emulsification process on sensory properties of compound Huangqi liquor

[0095]

[0096] Note: Different letters in the same column have significant difference (P < 0.05).

[0097] The present application takes the content determination of a variety of components such as astragaloside, hesperidin and taste score as indexes, investigates the influence of raw material selection, extraction process, yellow rice type and deployment technology on the quality of compound Huangqi liquor, and determines the best preparation process of compound Huangqi liquor. Finally, 3-year-old Shanxi native Huangqi and 5-year-old Xinhui dried orange peel are selected as the basic standard of raw materials for traditional Chinese medicine prescription. If Huangqi and dried orange peel from other places are selected, in addition to meeting the basic standard requirements of pharmacopoeia, the content of astragaloside needs to be referred to 3-year-old Shanxi native Huangqi, and the content of hesperidin needs to be referred to 5-year-old Xinhui dried orange peel. The content of effective components is not less than 90% of the standard. Through orthogonal experiment, the optimal alcohol extraction process conditions are determined, taking the extract yield and total flavonoid yield as indexes. Finally, the best extraction process is determined as alcohol extraction concentration 60%, extraction temperature 60℃, and extraction time 0.5h. The stability of compound Huangqi liquor is relatively intuitively represented by the milk separation index and sensory evaluation. Finally, millet yellow rice, 1 / 10 deployment concentration and high-speed homogenizer are selected for deployment of compound Huangqi liquor, and the quality of compound Huangqi liquor is more stable. In summary, when the extraction conditions are alcohol extraction concentration 60%, extraction temperature 60℃, extraction time 0.5h, and ethanol dosage 8 times, the content of effective components is higher when all traditional Chinese medicinal materials are co-extracted. The product obtained after deployment of millet yellow rice, 1 / 10 ratio and high-speed homogenization has stable process, and the appearance, taste and texture are the best.

[0098] III. The improvement effect of compound Huangqi liquor on lung function of pulmonary fibrosis rats: Idiopathic pulmonary fibrosis (IPF) is characterized by dry cough, dyspnea, and severe lung damage, with continuous deterioration of respiratory function. According to the clinical symptoms of idiopathic pulmonary fibrosis, traditional Chinese medicine classifies it into the category of "pulmonary atrophy" and "pulmonary obstruction", and the main pathological products are phlegm and blood stasis. The principle of diagnosis and treatment basically follows the treatment principles of regulating human qi, removing phlegm and blood stasis, and tonifying the lung and kidney to delay the progression of IPF. From the perspective of pharmacology, pulmonary fibrosis is a terminal change of a large class of pulmonary diseases characterized by fibroblast proliferation, massive extracellular matrix accumulation, and inflammation damage, and tissue structure destruction. That is, the normal alveolar tissue is damaged and abnormal repair leads to structural abnormalities. The pathological process of the bleomycin-induced pulmonary fibrosis model in rats is similar to that of human pulmonary fibrosis, and it is a classic animal model for studying pulmonary fibrosis. Today, the clinical treatment is mostly symptomatic treatment with glucocorticoids and immunosuppressive drugs, but the treatment effect is poor, and long-term use can cause serious dependence and adverse reactions.

[0099] The Huangqi in the present application tonifies the lung and spleen qi, and is the monarch drug for treating lung qi deficiency. The dried ginger disperses and warms to unblock, and enters both the blood and qi systems, so it is the minister drug. The perilla seed is used for descending, dispersing, and unblocking, and is the assistant drug. The tangerine peel unblocks lung qi, resolves phlegm and dampness, and dries dampness to tonify the spleen, so it is the ministerial drug. The present application studies the effect of the whole formula without perilla seed while studying the efficacy and mechanism of the compound Huangqi liquor in treating IPF, and explores the key role of perilla seed in the formula from the perspective of traditional Chinese medicine pharmacology.

[0100] 1. Experimental materials: Experimental animals: 80 SPF male SD rats, 4 weeks old, weighing 200-220 g, provided by Sperofree (Beijing) Biotechnology Co., Ltd., and raised in the general animal room. Experimental reagents: Compound Huangqi liquor (a compound preparation prepared from Huangqi, dried ginger, perilla seed, and tangerine peel), reduced flavor formula (a compound preparation prepared from Huangqi, dried ginger, and tangerine peel); Pirfenidone (produced by Beijing Contini Pharmaceutical Co., Ltd., national drug code: 0133376); bleomycin (Invitrogen, USA, batch number 5001-8); MM antibody, TIM antibody (Abcam, USA, batch numbers a8803 and a8978, respectively); ready-to-use immunohistochemical kit (Beijing Baoerleibo Technology Co., Ltd., batch number 20220330).

[0101] 2. Experimental method

[0102] (1) Animal grouping and its treatment: 100 healthy male rats (200g-220g) were selected, after one week of adaptive training, they were randomly divided into 5 groups according to the random number table method: blank group, model group, pirfenidone group, compound Huangqi medicinal liquor group and reduced flavor prescription group, 20 rats in each group. After the rats were anesthetized by intraperitoneal injection of 4% chloral hydrate, they were fixed on the experimental table in supine position, the neck was sterilized with alcohol and shaved, the skin of the neck was incised, the trachea was separated layer by layer and reserved. The blank group was injected with normal saline 3ml per kilogram of body weight into the trachea at one time. The model group, pirfenidone group, compound Huangqi medicinal liquor group and reduced flavor prescription group were injected with equal volume of bleomycin 4mg per kilogram of body weight into the trachea at one time. After operation, the animal state was closely observed, finally the rats were sutured and fed in the animal laboratory with suitable conditions. The next day after modeling, the rats were given gavage for 28 consecutive days.

[0103] (2) Sample collection and treatment: 5 rats were randomly selected from each group on the 7th, 14th, 21st and 28th day, and their lungs were taken after being sacrificed, and the double lungs were quickly immersed in 10% neutral formaldehyde for fixation. Paraffin sections were prepared through steps such as ethanol gradient dehydration and xylene transparency, and the fixed thickness of each section was 5μm. Ten copies of each tissue were taken for preservation for subsequent analysis.

[0104] (3) General observation of rats: During the experiment, the overall condition of the rats was observed, including mental state, fur color, body weight change, diet, drinking water, breathing, excretion, response ability to external stimuli, etc.

[0105] (4) Lung function detection of pulmonary fibrosis model rats: After the rats in each group were anesthetized by intraperitoneal injection of 4% chloral hydrate, they were fixed on the experimental table in supine position, the neck was sterilized with alcohol and shaved, the skin of the neck was incised, and the trachea was exposed by separating layer by layer. The trachea was cut open to a small opening, a cannula was inserted, and the rats were placed in the sealed cavity of the animal lung function analysis system, and the tracheal cannula was connected with the gas path of the cavity. The airway resistance and tidal volume of the rats were detected and recorded in real time by computer.

[0106] (5) Lung coefficient determination of pulmonary fibrosis model rats: After the rats completed the lung function detection, their lung tissues were taken after being sacrificed, washed several times with normal saline, and the surface moisture of the lung tissue was wiped dry with filter paper, then weighed and recorded, and the lung coefficient was calculated. Lung coefficient (%) = lung weight (g) / body weight (g) x 100.

[0107] (6) Histopathological observation of rat lung tissue: The lung tissue was collected to observe the lung tissue morphology, and the lung tissue was analyzed by hematoxylin-eosin (HE) and Masson staining, neutral gum sealing, air drying, and optical microscope observation of section staining.

[0108] (7) Immunohistochemical method for detecting the expression of lung tissue MMP-9, TIMP-1: Immunohistochemical method was used to determine the expression of lung tissue matrix metalloproteinase 9 (MMP-9) and matrix metalloproteinase tissue inhibitor-1 (TIMP-1). The cytoplasm and cell membrane were brownish yellow as positive cells, and the integral optical density value (IOD) of positive cells in the picture was analyzed under Olympus C3 microscope and Imag-Pro-Plus 6.0 software.

[0109] (8) Data processing: The data obtained by experiment was statistically analyzed by SPSS 7.0 software, and the data was expressed as mean ± standard deviation , single factor analysis of variance was used for comparison among multiple groups, and SNK-q test was used for further pairwise comparison, P<0.05 was considered statistically significant.

[0110] 3. Experimental results

[0111] (1) General observation results of rats: The rats in the blank group grew well, had good mental state, high reaction sensitivity, normal water intake, gradually increasing weight, shiny fur, normal breathing and excretion. After 1 day of modeling, except for the blank group, the rats in each group had weak breathing, little movement, poor spirit, dull fur, and reduced food and water intake. After 7 days, except for the blank group, the rats in each group had rapid shallow breathing, less food and water intake, and still thin body type, with decreased body weight. After 14 days, 21 days and 28 days, compared with the blank group, the rats in the model group had dull fur, poor spirit, little movement, reduced food and water intake, and significantly decreased body weight (P<0.05); compared with the model group, the rats in the pirfenidone and compound Huangqi medicinal wine group had good mental state, shiny fur, slightly rapid breathing, and significantly increased body weight (P<0.05). There was no significant difference in the rats in the reduced flavor compound Huangqi medicinal wine group within 7 days and 14 days, and the rats' mental state improved and the body weight increased significantly (P<0.05) after 21 days. The effect of compound Huangqi medicinal wine on the body weight of pulmonary fibrosis model rats is shown in Table 17.

[0112] Table 17: Effect of compound Huangqi medicinal wine on body weight of pulmonary fibrosis model rats

[0113]

[0114] Note: There was significant difference (P<0.05) among different letters in the same column. HQJ is compound Huangqi medicinal wine, and ASZ is compound Huangqi medicinal wine with reduced flavor.

[0115] (2) Effect of Compound Astragalus Wine on Lung Coefficient in Rats with Pulmonary Fibrosis: Compared with the blank group, the lung coefficient of the model group rats was significantly increased (P<0.05); compared with the model group, the lung coefficient of the pirfenidone group, the compound astragalus wine group, and the reduced-flavor formula group was significantly decreased (P<0.05); compared with the pirfenidone group, the compound astragalus wine group showed a significant difference in the lung coefficient of rats (P<0.05), but the reduced-flavor formula group showed no significant difference. The results are shown in Table 18.

[0116] Table 18: Effects of Compound Astragalus Wine on Lung Coefficient in Rats with Pulmonary Fibrosis

[0117]

[0118] Note: There are significant differences between different letters in the same column (P<0.05). HQJ is Compound Astragalus Wine, and ASZ is Compound Astragalus Wine with Perilla Seed Extract.

[0119] (3) Effects of Compound Astragalus Wine on Lung Function in Rats with Pulmonary Fibrosis: Compared with the blank group, the airway resistance of rats in the model group increased (P<0.05) and the tidal volume decreased (P<0.05). Compared with the model group, the airway resistance of rats in the pirfenidone group, the compound astragalus wine group, and the reduced-flavor formula group decreased significantly (P<0.05) and the tidal volume increased significantly (P<0.05). Compared with the pirfenidone group, the compound astragalus wine group had a significant effect (P<0.05) and was better than the pirfenidone group, while there was no difference in the reduced-flavor formula group. There was no significant difference in tidal volume between the compound astragalus wine group and the reduced-flavor formula group compared with the pirfenidone group. The results are shown in Table 19.

[0120] Table 19: Effects of Compound Astragalus Wine on Lung Function in Rats with Pulmonary Fibrosis Model

[0121]

[0122] Note: There are significant differences between different letters in the same column (P<0.05). HQJ is Compound Astragalus Wine, and ASZ is Compound Astragalus Wine with Perilla Seed Extract.

[0123] (4) Effects of compound Astragalus wine on non-tissue structures of a rat model of pulmonary fibrosis: HE staining results showed that the lung structure of the blank group rats was normal, the alveoli were intact and visible, no obvious inflammatory cell infiltration was observed, and almost no pathological changes were observed. Figure 1 A). The alveolar structure of the model group rats was disordered, with a large number of inflammatory cells infiltrating the interstitial area, and some collagen formed dense red collagen deposits, showing relatively obvious fibrotic lesions. Figure 1B, arrows indicate). Compared with the model group, the rats in the pirfenidone group, the compound astragalus medicinal liquor group and the reduced flavor group had less inflammatory cell infiltration in the lung tissue, and relatively less collagen fibers Figure 1 C). Masson staining results showed that the lung tissue structure of the blank group rats was normal, the alveoli were complete and clear, and there was no obvious inflammatory cell infiltration, and a small amount of blue collagen fiber deposition Figure 1 a, a small amount of fibrosis in the lung of the rat near the bronchus is a normal phenomenon). The lung alveolar structure of the model group rats was disordered, the lung tissue was severely damaged, there were a large number of inflammatory cell infiltrations, and a large amount of blue collagen fibers were deposited, and the fibrosis degree was serious Figure 1 b, arrows indicate). Compared with the model group, the rats in the pirfenidone group, the compound astragalus medicinal liquor group and the reduced flavor group had less inflammatory cell infiltration in the lung, and the degree of fibrosis was reduced Figure 1 c).

[0124] (5) Effect of Compound Astragalus Medicinal Liquor on MMP-9 Positive Cell Rate in Lung Tissue of Pulmonary Fibrosis Rats: Compared with the blank group, the positive cell rate of MMP-9 in the lung tissue of the model group rats was significantly increased at four time nodes (P<0.05), indicating that the pulmonary fibrosis model of rats in this experiment was successfully established; Compared with the model group, the positive cell rate of MMP-9 in the lung tissue of rats in the pirfenidone group, the compound astragalus medicinal liquor group and the reduced flavor group was significantly reduced (P<0.05); Compared with the pirfenidone group, the positive cell rate of MMP-9 in the lung tissue of rats in the compound astragalus medicinal liquor group and the reduced flavor group had no significant difference within 7 days and 14 days of intervention, and after 21 days of intervention, the positive cell rate of MMP-9 in the lung tissue of the pulmonary fibrosis rat model in the compound astragalus medicinal liquor group was significantly reduced (P<0.05), but there was no significant difference in the reduced flavor group, indicating that the compound astragalus medicinal liquor had the effect of regulating the protein expression of MMP-9 in the lung tissue, thereby reducing the damage to the basement membrane of lung epithelial cells and participating in the repair of pulmonary fibrosis. The results are shown in Table 20.

[0125] Table 20: Effect of Compound Astragalus Medicinal Liquor on MMP-9 Positive Cell Rate in Lung Tissue of Fibrosis Model Rats

[0126]

[0127] Note: There was a significant difference (P<0.05) among different letters in the same column. HQJ is compound astragalus medicinal liquor, and ASZ is compound astragalus medicinal liquor with reduced flavor.

[0128] (6) The effect of compound Huangqi liquor on the positive cell rate of TIMP-1 in lung tissue of pulmonary fibrosis rats: Compared with the blank group, the positive cell rate of TIMP-1 in lung tissue of rats in the model group was significantly increased at four time nodes (P<0.05), indicating that the rat pulmonary fibrosis model was successfully established; compared with the model group, the positive cell rate of TIMP-1 in lung tissue of rats in the pirfenidone group and the compound Huangqi liquor group was significantly reduced (P<0.05); compared with the pirfenidone group, there was no significant difference in the positive cell rate of TIMP-1 in lung tissue of rats in the compound Huangqi liquor group and the reduced flavor group within 7 days, 14 days and 21 days of intervention, and after 28 days of intervention, the positive cell rate of TIMP-1 in lung tissue of pulmonary fibrosis rats in the compound Huangqi liquor group was significantly reduced (P<0.05), but there was no significant difference in the reduced flavor group, indicating that the compound Huangqi liquor group inhibited the expression of TIMP-1. The results are shown in Table 21.

[0129] Table 21: Effect of compound Huangqi liquor on the positive cell rate of TIMP-1 in lung tissue of fibrosis rats

[0130]

[0131] Note: There was a significant difference (P<0.05) between different letters in the same column. HQJ is compound Huangqi liquor, and ASZ is compound Huangqi liquor with reduced flavor of Suzi.

[0132] After bleomycin established the pulmonary fibrosis rat model, the rats had weak breathing, less movement, poor spirit, dull fur, reduced food and water intake, and weight loss. After intragastric administration of compound Huangqi liquor, the mental status of rats was improved, the fur was relatively lustrous, the food and water intake were increased, and the body weight was significantly increased, but the rats intragastrically administered with reduced flavor had no obvious improvement effect on pulmonary fibrosis in rats compared with those intragastrically administered with compound Huangqi liquor.

[0133] The FinePointe RC system is used for lung function determination of the pulmonary fibrosis model rats, and the results show that the airway resistance of the model group rats is significantly increased, and the tidal volume is obviously reduced. After giving the compound Huangqi medicinal liquor, the lung function of the pulmonary fibrosis model rats is significantly improved, and compared with the positive control group, the airway resistance is significantly reduced, and the tidal volume is significantly increased, and the results show that the effect is better than that of the positive control group. The lung coefficient is an important index reflecting the degree of pulmonary fibrosis of the pulmonary fibrosis model rats, and the experimental results show that the lung coefficient of the model group rats is significantly increased, and compared with the model group and the positive control group, the lung coefficient of the compound Huangqi medicinal liquor group rats is significantly reduced. Research shows that with the development of pulmonary fibrosis, fibrous connective tissue proliferation and large amount of collagen deposition cause the increase of lung weight, thereby causing the increase of lung coefficient. MMP-9 and TIMP-1 play an important role in the metabolism of extracellular matrix (ECM), and as a marker of airway fibrosis, the excessive production of MMP-9 and TIMP-1 in IPF patients may play a role in destroying the basement membrane and causing fibroblasts to invade the alveolar cavity to cause pulmonary fibrosis. The experimental results show that the compound Huangqi medicinal liquor group significantly reduces the positive cell rate of MMP-9 and TIMP-1, indicating that MMP-9 and TIMP-1 play an important role in the process of alveolar inflammation, which can reduce airway inflammation and improve the effect of pulmonary fibrosis airway remodeling. In summary, based on the evaluation of the lung function of the compound Huangqi medicinal liquor on the pulmonary fibrosis model rats, the results show that the compound Huangqi medicinal liquor has an improving effect on the general condition of the pulmonary fibrosis model rats, increases the body weight of the rats, reduces the airway resistance and increases the tidal volume to improve the lung function of the rats, reduces the organ weight to reduce the lung coefficient index of the rats, and reduces the positive cell rate of MMP-9 and TIMP-1 to improve pulmonary fibrosis.

[0134] IV. Improvement of Compound Huangqi Liquor on Cognitive Function of Diabetic Rats: In recent years, the number of people with diabetes has increased dramatically. Long-term chronic hyperglycemia affects the function of important organs throughout the body, especially in the brain, including learning, attention, memory, calculation, language, and executive function. According to research, long-term use of insulin can cause metabolic disorders, allergic reactions, and hypoglycemia. Traditional Chinese medicine classifies cognitive impairment as "not smart, stupid, and forgetful." Diabetic cognitive impairment is one of the chronic complications, and the pathogenesis is primarily due to spleen deficiency. Spleen deficiency leads to poor digestion and absorption of food and fine substances, and the brain loses nutrition, resulting in impaired cognitive function. The principle of diagnosis and treatment is to supplement the spleen and stomach. The spleen and stomach are the foundation of the brain. A strong spleen and stomach can produce blood and nourish the brain, and the brain is clear. Huangqi Decoction can supplement the spleen and lung, promote the circulation of qi and blood, and reduce the side effects of insulin. The application studies the effect of Suzi Decoction on diabetic cognitive impairment and explores the key role of Suzi in the prescription from the perspective of traditional Chinese medicine pharmacology.

[0135] 1. Experimental materials: Experimental animals: 80 SPF male SD rats, 4 weeks old, weighing 200-220 g, provided by Sperof (Beijing) Biotechnology Co., Ltd., and raised in the general animal room. Experimental reagents: Compound Huangqi Liquor (a compound preparation prepared from four traditional Chinese medicines: Huangqi, Ganjiang, Suzi, and Chenpi), Suzi Decoction (a compound preparation prepared from three traditional Chinese medicines: Huangqi, Ganjiang, and Chenpi), Metformin Hydrochloride Tablets (batch number 121209, Tianjin Pacific Pharmaceutical Co., Ltd.), Streptozotocin (Bao Biological Engineering Dalian Co., Ltd., batch number 040357), Phosphate Buffer Saline (PBS), BDNF rabbit antibody, goat anti-rabbit secondary antibody (Shanghai Sixin Biotechnology Co., Ltd., batch number 012K, 205S), avidin-biotin-peroxidase complex kit (Vector Laboratories, USA, batch number S023), DAB color reagent kit (Wuhan Dr. De Biological Engineering Co., Ltd., batch number 190107), and 3,3-diaminobenzidine tetrahydrochloride reagent (Shanghai Beinuo Biotechnology Co., Ltd., batch number 190314).

[0136] 2. Experimental method

[0137] (1) Animal grouping and its treatment: 50 healthy male rats, 135g-165g, were selected after one week of adaptive training. Except for the blank group, the rats in other groups were fed with high-fat and high-sugar feed for one month. After one month, the blank group was injected with citric acid buffer, and the rats in other groups were injected with streptozotocin (STZ) intraperitoneally. 24h later, the tail tip venous blood was collected, and the fasting blood glucose of the rats was measured by a portable blood glucose meter. The blood glucose was greater than 16.7mmol·l -1 and the cognitive impairment rats were screened by the jumping platform experiment. The jumping platform experiment was performed for 3 days, twice a day. By comparing with the blank group, the average latency was significantly reduced, and it was determined that the modeling was successful, and the high-fat and high-sugar feed was continued for 2 months.

[0138] The rats with successful modeling were randomly divided into model group, metformin hydrochloride group, compound radix astragali medicinal liquor group and reduced flavor prescription group by random number table method, 10 rats in each group. The blank group and the model group were given the same volume of normal saline by gavage, and the metformin hydrochloride group, the compound radix astragali medicinal liquor group and the reduced flavor prescription group were given a certain amount of metformin hydrochloride, compound radix astragali medicinal liquor and compound radix astragali medicinal liquor reduced flavor prescription by gavage, and the gavage was continued for 2 months. The mental state, food and water intake of rats were observed daily, and the Morris water maze experiment was performed at the end of the period.

[0139] (2) Sample collection and its treatment: After the Morris water maze experiment, the rats were fasted for 12h, 3 rats in each group were taken their brain tissues, and were immersed in 10% neutral formaldehyde for fixation. Through the steps of ethanol gradient dehydration and xylene transparency, paraffin sections were prepared, and the fixed thickness of each section was 4μm.

[0140] (3) General condition observation: The diet, behavior, water intake, mental state, activity state, hair luster and other conditions of rats in each group were observed. The fasting blood glucose of rats was tested by blood glucose meter and test strip at the tail tip, once every 2 weeks, and the last fasting blood glucose determination was performed. The rats were fasted for 10h the night before the body weight determination, and were placed on an electronic balance to measure the body weight, which was repeated for 3 times, and the data was recorded.

[0141] (4) Morris water maze test: After the end of feeding, the spatial cognitive and memory ability of mice was tested by Morris water maze training. The water maze consisted of a diameter of 150 cm, a height of 55 layers, and a water filled with 25 cm, which was divided into four quadrants (divided into east, south, west, north area), and a solid underwater escape platform with movable position was built in. The mice were randomly placed in one quadrant and entered the water in a back projection mode, and found the underwater escape platform. Each training was 120 s, and the training ended when the mouse found the platform. If the mouse did not find the escape platform within 120 s, it was taken out and placed on the platform for 60 s before the next experiment. The time for the mouse to find the escape platform (escape latency) and the distance of swimming (escape distance) were recorded. After 4 days of training, the experiment test was carried out on the fifth day, and the escape platform was removed. After the mouse was put into the water, the number of times of crossing the original platform and the time of staying in the original platform area within 120 s were recorded.

[0142] (5) Immunohistochemical detection of BDNF protein expression in diabetic model rats: The protein expression level of brain-derived neurotrophic factor (BDNF) in rat brain tissue was determined by immunohistochemical staining. The rat hippocampal tissue section was deparaffinized and rehydrated, then antigen repair was performed, 10% normal goat serum PBS was added dropwise, and the solution was treated at room temperature for 1 h, and BDNF rabbit antibody was incubated at 4°C for 48 h. Dropwise addition of secondary antibody (7.5 mg / ml biotinylated goat anti-rabbit) was treated at room temperature for 3 h, and anti-biotin protein biotin peroxidase compound was incubated at room temperature for 1 h, and 3,3-diaminobenzidine tetrahydrochloride reagent was colored at room temperature for 20 min, and hematoxylin was re-stained for 30 s, and gradient ethanol was dehydrated and dried, xylene was transparent, and neutral gum was sealed. Under a microscope at 400 times, the number of cells and the number of positive cells were counted and the average values were calculated. Under the Olympus C3 microscope, the integral optical density value (IOD) of the positive cells in the picture was analyzed by Imag-Pro-Plus 6.0 software.

[0143] (6) Data statistics: The data obtained from the experiment were statistically analyzed by SPSS7.0 software, and the data were expressed as mean ± standard deviation. Single factor analysis of variance was used for comparison among multiple groups, and SNK-q test was used for further pairwise comparison. P<0.05 was considered statistically significant.

[0144] 3、Experimental results

[0145] (1) General situation of rats: the rats in the blank group grew well, with bright fur, frequent activity, and normal water and food intake. Compared with the blank group, the rats in the high-fat feeding model group had significantly decreased body weight and significantly increased blood glucose (P<0.05), oily and easy-to-shed fur, decreased luster, listless spirit, and polyuria. Compared with the model group, the rats in the metformin group and the reduced-flavor group had no significant difference in body weight (P>0.05), and the rats in the compound Huangqi medicinal wine group had a reduced decrease in body weight (P<0.05). Compared with the model group, the rats in the metformin group, the compound Huangqi medicinal wine group, and the reduced-flavor group had significantly decreased blood glucose (P<0.05), and there was no significant difference between the compound Huangqi medicinal wine group and the reduced-flavor group (P>0.05). The results of the effects of the compound Huangqi medicinal wine on the body weight and blood glucose of the diabetic model rats are shown in Table 22.

[0146] Table 22: Effects of the compound Huangqi medicinal wine on the body weight and blood glucose of the diabetic model rats

[0147]

[0148] Note: There was a significant difference (P<0.05) between different letters in the same column. HQJ refers to the compound Huangqi medicinal wine, and ASZ refers to the compound Huangqi medicinal wine with reduced flavor of Suzi.

[0149] (2) Effects of the compound Huangqi medicinal wine on the Morris water maze behavior of diabetic rats: During the continuous training process, compared with the blank group, the rats in the model group had significantly prolonged escape latency on the third and fourth days (P<0.05), and significantly decreased number of platform crossings and target area residence time (P<0.05). Compared with the model group, the rats in the metformin group, the compound Huangqi medicinal wine group, and the reduced-flavor group had significantly shortened escape latency (P<0.05), and significantly increased number of platform crossings and target area residence time (P<0.05). On the fourth day of training, compared with the metformin group, the rats in the compound Huangqi medicinal wine group had significantly different escape latency, number of platform crossings, and target area residence time (P<0.05), but the reduced-flavor group had no significant difference, and the improvement of diabetic cognitive impairment was better than that of the metformin group and the reduced-flavor group. The results are shown in Tables 23 and 24.

[0150] Table 23: Effects of the compound Huangqi medicinal wine on the escape latency of diabetic rats in the Morris water maze

[0151]

[0152] Note: There was a significant difference (P<0.05) between different letters in the same column. HQJ refers to the compound Huangqi medicinal wine, and ASZ refers to the compound Huangqi medicinal wine with reduced flavor of Suzi.

[0153] Table 24: Effect of Compound Huangqi Medicinal Liquor on Spatial Exploration of Morris Water Maze in Diabetic Rats

[0154]

[0155] Note: Different letters in the same column have significant difference (P<0.05), HQJ is Compound Huangqi Medicinal Liquor, and ASZ is Compound Huangqi Medicinal Liquor Suzi Decoction.

[0156] (3) Effect of Compound Huangqi Medicinal Liquor on BDNF Protein Expression in Brain Tissue of Diabetic Rats: Compared with the blank group, the BDNF protein expression in brain tissue of the model group was significantly reduced (P<0.05); compared with the model group, the BDNF protein expression in brain tissue of the metformin group, the Compound Huangqi Medicinal Liquor group and the Suzi Decoction group was significantly increased (P<0.05); compared with the metformin group, the BDNF protein expression in brain tissue of the Compound Huangqi Medicinal Liquor group and the Suzi Decoction group was significantly higher than that of the metformin group (P<0.05). The results are shown in Table 25.

[0157] Table 25: Effect of Compound Huangqi Medicinal Liquor on BDNF Protein Expression in Brain Tissue of Diabetic Rats

[0158]

[0159] Note: Different letters in the same column have significant difference (P<0.05), HQJ is Compound Huangqi Medicinal Liquor, and ASZ is Compound Huangqi Medicinal Liquor Suzi Decoction.

[0160] After the modeling of the diabetic model rats by high-fat high-sugar feed combined with STZ injection, the body weight decreased, the blood sugar increased, the hair was oily and easy to fall off, the luster decreased, the spirit was debilitated, and was accompanied by polyuria symptoms. After intragastric administration of compound Huangqi medicinal liquor, the body weight of rats increased significantly, the blood sugar decreased, the hair was oily and easy to fall off, the mental state improved, and the polyuria symptoms were reduced. Compared with the compound Huangqi medicinal liquor, the reduced flavor did not improve all the symptoms. Morris water maze is the most commonly used experimental method for evaluating animal spatial learning and cognitive ability, including space navigation and space exploration. In the space exploration experiment, escape latency was selected as the evaluation index. In the space exploration experiment, the number of crossing the platform and the residence time in the target area were selected as the evaluation index. The experimental results showed that during the continuous training process, there was a statistically significant difference between the groups. Compared with the model group, the metformin group, the compound Huangqi medicinal liquor group and the reduced flavor group could shorten the escape latency of rats, significantly increase the number of crossing the platform and the residence time in the target area. Compared with the metformin group, there was a significant difference in the escape latency, the number of crossing the platform and the residence time in the target area of the rats in the compound Huangqi medicinal liquor group, and the improvement of cognitive impairment in diabetic rats was better than that in the metformin group and the reduced flavor group. Brain-derived neurotrophic factor (BNDF) is an important biological link between thought, emotion and movement, which can strengthen the internal connection of hippocampus and promote the growth of nerve cells, and is closely related to learning and memory function. The experimental results showed that compared with the model group, there was a statistically significant difference in the expression of BDNF protein in the brain tissue of rats in the metformin group, the compound Huangqi medicinal liquor group and the reduced flavor group, and there was a significant difference between the metformin group and the compound Huangqi medicinal liquor group and the reduced flavor group. There was no significant difference between the compound Huangqi medicinal liquor group and the reduced flavor group, and the effect was significantly better than that of the metformin group. This experiment confirmed that compound Huangqi medicinal liquor can regulate the progress of cognitive impairment, thereby improving cognitive and learning and memory ability, and is consistent with the clinical research that the expression level of BNDF protein is closely related to the condition of cognitive function of patients. In summary, after the treatment of the compound Huangqi medicinal liquor group for the diabetic cognitive dysfunction model rats, the body weight of rats increased significantly, the blood sugar decreased, the hair was oily and easy to fall off, the mental state improved, and the polyuria symptoms were reduced. Through the evaluation of Morris water maze experiment, it was found that the spatial learning and cognitive ability of rats was improved to some extent.

[0161] Five, the improvement effect of compound Huangqi liquor on osteoporosis model rats: osteoporosis is mainly characterized by increased bone fragility and easy fracture. For the pathogenesis of osteoporosis, TCM has not formed a unified view, but it is generally believed that kidney deficiency is the root, liver deficiency is the key factor, and spleen deficiency is an important factor, among which blood stasis plays a promoting role. It is believed that osteoporosis such as "bone atrophy", "bone numbness", "bone fracture", "symptoms before and after menopause", "asthenia" and other diseases, which are clinically manifested as back pain, limb weakness, bone deformation, and soreness of the waist and knees. Modern medical research shows that kidney essence deficiency is the root, spleen deficiency is the inducement, and blood stasis is the promoting factor. Therefore, the diagnosis and treatment of TCM should be to invigorate the spleen to treat the root and promote blood circulation to treat the symptoms, that is, the spleen is strong, the kidney essence is sufficient, the collaterals are unobstructed, and the blood is vigorous, so the muscles and bones are strong. It provides the required nutrients for bone development and bone regeneration and repair, and plays an important role in reducing the symptoms of osteoporosis.

[0162] Huangqi in this prescription is good for tonifying the spleen and indirectly promoting blood circulation as the monarch drug. Ganjiang is pungent and warm, and is widely used for blood stasis and qi stagnation. It is used as the minister drug for dispelling wind-cold and dampness. Suxin is warm and moistening, and is used as the assistant drug for treating chronic cough and asthma. Chenpi is pungent, bitter, warm and dry, and is used as the guide drug for regulating qi and invigorating the spleen. This study not only studies the effect of compound Huangqi liquor on osteoporosis, but also studies the effect of Suxin reduced formula, to explore the key role of Suxin in the prescription from the perspective of traditional Chinese medicine pharmacology.

[0163] 1. Experimental materials: Experimental animals: 32 SPF male SD rats, weighing (150-200) g, provided by Sibeifu (Beijing) Biotechnology Co., Ltd., and raised in the general animal room. Experimental drugs: Compound Huangqi liquor (a compound preparation prepared from four traditional Chinese medicines: Huangqi, Ganjiang, Suxin and Chenpi), reduced formula (a compound preparation prepared from three traditional Chinese medicines: Huangqi, Ganjiang and Chenpi), Xianling Gubao capsules (Tongji Pharmaceutical Co., Ltd., National Drug Code Z20025337); TRAP staining kit (Sigma AIdrich, Germany, batch number P114); p38 MAPK, JNK, ELISA kit (Shanghai Zymed Biotech Co., Ltd., batch numbers m37037, m20318 and m37375); interleukin (IL-1) (Shanghai Bangyi Biotechnology Co., Ltd., batch number S16.sV9); hematoxylin and eosin staining solution (Shanghai Wei'ao Biotechnology Co., Ltd., batch numbers 00501 and 00702).

[0164] 2. Experimental method

[0165] (1) Animal grouping and treatment: 50 three-month-old male Wistar rats, weighing about 150 g. After adaptive feeding for 1 week, they were randomly divided into blank group, control group, dexamethasone group, compound Huangqi medicine treatment group and Jianwei prescription group, 10 in each group, for 12 weeks. The first 8 weeks: the rats in the control group, dexamethasone group, compound Huangqi medicine treatment group and Jianwei prescription group were injected with dexamethasone 1 mg / kg intramuscularly, twice a week. The blank group was injected with the same amount of isotonic saline intramuscularly. The indicators for judging whether the modeling was successful were: serum bone metabolism markers, bone microstructure and bone density determination. In this experiment, serum bone metabolism markers and bone density showed that the modeling was successful. The last 4 weeks: the control group was given Xianling Gubao capsules 30.83 mg / 100 g subcutaneously once a day, the compound Huangqi medicine treatment group was given compound Huangqi medicine by gavage, and the Jianwei prescription group was given compound Huangqi Jianwei prescription by gavage, once every 2 weeks. The blank group and the dexamethasone group were not given drugs. All animals were raised under the same conditions, with free access to food and water, and their body weight was measured once a week.

[0166] (2) Sample collection and processing: After feeding, the rats in each group were deprived of food but not water for 12 hours the night before. The next morning, after being anesthetized with ether, they were weighed and numbered, their eyeballs were removed for blood collection, and the upper serum was taken after centrifugation and stored in a -20°C refrigerator. At the same time, the rats were sacrificed by cervical dislocation, and their liver, spleen and kidney were removed, washed several times with physiological saline, and then weighed on an electronic balance after the water was absorbed with filter paper. Then the left and right femurs of the rats were removed, the surrounding soft tissue was removed, and they were immersed in 10% neutral formaldehyde for fixation. Paraffin sections were prepared by ethanol gradient dehydration, xylene transparency and other steps, with a fixed thickness of 4 μm per section.

[0167] (3) General observation of rats: During the administration period, the activity of the rats in each group before and after administration was observed at regular intervals every day, including food and water intake, stool condition, whether the hair was shiny or not, whether there were abnormal activities and emotional performance, and the body weight of the rats in each group was measured at a certain time, the data was recorded and compared and analyzed.

[0168] (4) TRAP staining method for detecting the number of bone tissue and positive osteoclasts in osteoporosis model rats: the bone marrow cavity bone surface was washed with a-MEM and penicillin 1:10 solution to show white, and the extract was blown and extracted. Centrifuge the extract at 3200 r / min for 5 min, discard the supernatant, and drop a-MEM culture solution 8 mL. After blowing and mixing thoroughly, 0.5 mL of the suspension was dropped into a culture dish containing a cover glass, and incubated at 25°C for 30 min. Select a 24-well culture plate, take 4 holes and divide them into 4 groups, respectively, blank group, control group, Xianling Gubao capsule group, compound Huangqi medicinal wine group and reduced flavor group. After 10 days of culture, the cover glass was taken out, TRAP staining agent was added, and incubated at 37°C for 50 min. PBS buffer was added, xylene was transparent, and neutral gum was sealed. Under Olympus C3 microscope, take pictures, and take the purple red cells with more than 3 bubbles and large cell nucleus as osteoclasts. The area ratio of positive cells in the picture was analyzed by Imag-Pro-Plus 6.0 software, and the mean value of 3 non-adjacent fields was selected.

[0169] (5) ELISA detection of serum JNK, p38 MAPK and IL-1 content: enzyme-linked immunosorbent assay (ELISA) was used to detect the content of c-Jun amino terminal kinase (JNK), 8 mitogen-activated protein kinase (p38 MAPK) and interleukin-1 (IL-1) in rat serum. Set standard sample hole, sample hole and blank hole, standard sample hole add different solubility of control product 50uL, sample hole add sample diluent and sample in turn, blank hole except. After sealing the plate with sealing film, incubate at 37°C for 60 min. Dilute the 20-fold concentrated washing solution with distilled water 20 times for standby. Remove the sealing film, discard the liquid, and dry. Add washing solution to each hole, stand for 30 s, then discard. Repeat 5 times, and dry. Add color developing agent A 50uL to each hole, then add color developing agent B 50uL, mix gently, and develop color at 37°C for 15 min. Add stop solution 50uL to each hole, stop the reaction (at this time the blue color turns yellow), then measure OD450 value within 5 min. Use professional software to analyze the standard sample concentration.

[0170] (6) Data statistics: the data obtained by experiment were statistically analyzed by SPSS7.0 software, and the data were expressed as mean ± standard deviation (x±s) ) of mean ± standard deviation (x±s). Single factor analysis of variance was used for comparison among multiple groups, and SNK-q test was used for further pairwise comparison. P<0.05 was considered statistically significant.

[0171] 3、Experimental results

[0172] (1) Observation results of general condition of rats: The rats in the blank group were growing well, in good spirits, highly responsive, gradually increasing in weight, with glossy fur, and no other obvious abnormalities. One week after modeling, except for the blank group, the rats in each group had weak breathing, little movement, poor spirits, dull fur, and reduced food and water intake. Two weeks after administration, compared with the blank group, the rats in the control group had poorer spirits, dull fur, occasional hair loss, and significantly lower weight (P<0.05); compared with the control group, in the second and tenth weeks, the rats in the Xianling Gubao capsule group, the compound astragalus wine group, and the reduced-flavor formula group had good spirits, glossy fur, and slightly decreased weight. In the twelfth week, there was a significant difference between the compound astragalus wine group and the reduced-flavor formula group (P<0.05); compared with Xianling Gubao capsule, in the twelfth week, the compound astragalus wine group had a significant difference (P<0.05), and the rat weight showed an upward trend, while there was no significant difference in the reduced-flavor formula group. The results are shown in Table 26.

[0173] Table 26: Weight Changes in Osteoporosis Model Rats in Each Group

[0174]

[0175] Note: There are significant differences between different letters in the same column (P<0.05). HQJ is Compound Astragalus Wine, and ASZ is Compound Astragalus Wine with Perilla Seed Extract.

[0176] Effects of Compound Astragalus Wine on Bone Morphology and Osteoclast Number in Osteoporosis Model Rats: TRAP staining results showed that the trabeculae in the blank group were intact without obvious fractures, the medullary cavity was normal, and the osteocytes were evenly arranged. Figure 2 A); In the control group of osteoporosis model rats, the trabecular bone morphology was blurred and extensively fractured, the medullary cavity was widened, and the osteocytes were scattered. Figure 2 B, as indicated by the arrow); compared with the control group, the Xianling Gubao capsule group, the compound astragalus medicinal wine group, and the reduced-flavor formula group showed improved trabecular morphology and more compact structure, reduced medullary cavity, more uniform periosteal arrangement, and inhibition of osteoclast formation. Figure 2 C). The osteoclast counts of rats in each group are shown in Table 27. Compared with the blank group, the number of osteoclasts in the control group was significantly increased (P < 0.05). Compared with the control group, the number of osteoclasts in the Xianling Gubao capsule group, the compound astragalus wine group, and the reduced-flavor formula group was significantly decreased (P < 0.05). Compared with the Xianling Gubao capsule group, the number of osteoclasts in the compound astragalus wine group was significantly different and showed a decreasing trend (P < 0.05), but there was no significant difference in the reduced-flavor formula group.

[0177] Table 27: Osteoclast counts in rats of each group

[0178]

[0179] Note: the same column within different letters have significant difference (P<0.05), HQJ is compound Huangqi medicine, ASZ is compound Huangqi medicine Suzi reduced flavor.

[0180] (3) The influence of compound Huangqi medicine on the content of JNK, p38 MAPK and IL-1 in serum of osteoporosis model rats: the results are shown in Table 28, compared with the blank group, the content of p38 MAPK, JNK and IL-1 in serum of the control group rats is significantly increased (P<0.05); compared with the control group, the content of p38 MAPK, JNK and IL-1 in serum of the Xianling Gubao capsule group, compound Huangqi medicine group and reduced flavor group is significantly reduced (P<0.05); compared with the Xianling Gubao capsule group, the content of JNK in serum of the compound Huangqi medicine group and reduced flavor group has no significant difference, the content of p38 MAPK and IL-1 has significant difference, and is lower than that of the Xianling Gubao capsule group.

[0181] Table 28: the content of JNK, p38 MAPK and IL-1 in serum of rats

[0182]

[0183] Note: the same column within different letters have significant difference (P<0.05), HQJ is compound Huangqi medicine, ASZ is compound Huangqi medicine Suzi reduced flavor.

[0184] The present application adopts the classic modeling method of dexamethasone-induced osteoporosis in rats, simulates the osteoporosis state of middle-aged and elderly people, and after the modeling of rats, the state is poor, the breathing is weak, the activity is less, the hair color is dull, and the body weight is significantly decreased. After intragastrically administering the compound Huangqi medicine, the mental state of the rats is recovered, the hair color is lustrous compared with before, the body weight decrease is reduced, compared with the rats intragastrically administered with the compound Huangqi medicine, the rats in the reduced flavor group are not completely improved. The number, quality and morphology of bone trabecula are one of the important markers of bone quality, and the bone trabecula can reflect the severity of osteoporosis. The present application proves that the bone trabecula morphology of rats in the osteoporosis state is blurred, widely broken, the bone marrow cavity is widened, and the bone cells are arranged in disorder. After intragastrically administering the compound Huangqi medicine and the reduced flavor group, the bone trabecula morphology is improved compared with the model group, the structure is more compact, the bone marrow cavity is reduced, and the bone quality is improved, which has the effect of preventing and treating osteoporosis. In summary, after the compound Huangqi medicine group is administered to the osteoporosis model rats, the body weight decrease of the rats is relieved, the activity is increased, the mental state is improved, and the content of p38 MAPK, JNK and IL-1 in serum of the rats is reduced by ELISA method, and the rats with osteoporosis are improved.

Claims

1. A compound Huangqi medicinal liquor, characterized in that: Low degree millet yellow wine as the base wine, Astragalus, dried ginger, perilla fruit, dried tangerine or orange peel are crushed, 65 ° liquor is used to immerse the medicine, 120 r / min oscillation extraction, the extract is concentrated and then added to the yellow wine to mix and homogenize; among them: Astragalus 12g, dried ginger 9g, perilla fruit 9g, dried tangerine or orange peel 6g; ​ The specific preparation method comprises the following steps: (1) raw material crushing: Astragalus, dried ginger, perilla fruit, dried tangerine or orange peel are crushed to 60 mesh; (2) medicine extraction: the medicine powder obtained in step (1) is added to 65 ° liquor to immerse the medicine, the liquor dosage is 8 times the dosage of the medicine powder; the extraction temperature is controlled at 60 °C, and the extraction is carried out for 0.5 h; after the extraction is completed, heating is opened to concentrate to 1 / 4 to 1 / 3 of the original volume; (3) mixing: the medicine concentrated liquid obtained in step (2) is added to millet yellow wine, and the medicine extract accounts for 1 / 5 to 1 / 20 of the volume of the final product based on the final product as the calculation reference; (4) homogenization: after mixing, it is placed in a high-speed homogenizer, homogenized at 5000 rpm / min for 8 min, and then stirred in a blender at 18000 r / min for 10 min to mix evenly; The Astragalus is 3-year-old Shanxi Astragalus or 1-year-old Heilongjiang Astragalus; the dried tangerine or orange peel is 5-year-old Xinhui dried tangerine or orange peel.

2. The compound Huangqi medicinal liquor according to claim 1, characterized in that: The medicine concentrated liquid is added to millet yellow wine, and the medicine extract accounts for 1 / 5, 1 / 10 or 1 / 20 of the volume of the final product based on the final product as the calculation reference.

3. The compound Huangqi medicinal liquor according to claim 1, characterized in that: The medicine concentrated liquid is added to millet yellow wine, and the medicine extract accounts for 1 / 10 of the volume of the final product based on the final product as the calculation reference.

4. The application of the compound Astragalus medicinal liquor of claim 1 in the preparation of a medicine for treating pulmonary fibrosis.

5. The application of the compound Astragalus medicinal liquor of claim 1 in the preparation of a medicine for treating cognitive impairment caused by diabetes.

6. The application of the compound Astragalus medicinal liquor of claim 1 in the preparation of a medicine for treating osteoporosis.

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