Traditional Chinese medicine composition for treating kidney-yang deficiency and having antioxidant function as well as preparation method and application of traditional Chinese medicine composition

Abstract

The invention provides a traditional Chinese medicine composition for treating kidney-yang deficiency and having an anti-oxidation function as well as a preparation method and application thereof, and belongs to the technical field of traditional Chinese medicines. The traditional Chinese medicine composition comprises the following raw material medicines in parts by mass: 2-5 parts of rhizoma polygonati, 2-5 parts of raspberry, 2-5 parts of cistanche, 2-5 parts of eucommia male flower and 1-3 parts of cordyceps sobolifera sporocarp. The traditional Chinese medicine composition disclosed by the invention is relatively simple in formula, not only has the effect of treating kidney-yang deficiency, but also has an anti-oxidation effect, and can prevent and treat diseases related to oxidative stress through an anti-oxidation way on the basis of improving the symptom of kidney-yang deficiency.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine, and in particular relates to a traditional Chinese medicine composition for treating kidney yang deficiency and having antioxidant function, as well as a preparation method and application thereof. Background Art

[0002] Kidney yang deficiency is an important syndrome in traditional Chinese medicine theory. It is characterized by deficiency of kidney yang qi and decreased warming function. Clinical manifestations include soreness of waist and knees, fear of cold limbs, lack of energy, pale and fat tongue with white fur, and weak pulse. It is manifested in diseases of multiple systems such as respiratory, circulatory, endocrine, and digestive systems. With the in-depth research and clinical exploration of kidney yang deficiency, scientifically significant modeling methods and evaluation criteria have been formed. However, the current therapeutic drugs or methods for kidney yang deficiency syndrome still have certain limitations, and the improvement effect on the various complex symptoms caused by kidney yang deficiency needs to be further improved. In addition, some drugs may have certain side effects or inadequate efficacy.

[0003] In addition, in modern medical research, oxidative stress is considered to be closely related to aging and various chronic diseases. Excess free radicals can cause damage to various biomacromolecules such as DNA, proteins, and lipids, which in turn affects gene expression and regulatory systems, changes the differentiation state of cells, and ultimately leads to aging changes. This process also plays an important role in the pathogenesis of many chronic diseases such as neurodegenerative diseases (Alzheimer's disease, Parkinson's disease), cardiovascular diseases, and diabetes. At present, although it has been recognized that antioxidants have potential value in preventing and treating these diseases and delaying aging, existing antioxidant drugs or health products still have many problems in terms of efficacy, safety, and diversity of targets. It is of great significance to develop natural drugs or preparations with high-efficiency antioxidant effects and comprehensive regulation of body functions.

[0004] Currently, regarding drugs for treating kidney yang deficiency, there are patents CN110051784A - a new application of a Chinese medicine composition in the preparation of drugs for treating and preventing kidney yang deficiency, the formula of which is: 8-15 parts of Chinese angelica sinensis, 2-5 parts of Codonopsis pilosula, 2-5 parts of Polygonatum sibiricum, 2-5 parts of Aucklandia lappa and 0.1-0.5 parts of vitamin C; patent CN108578573A - a composition that has the effects of tonifying kidney yang, enhancing immunity and relieving fatigue, the formula of which is a water extract of Astragalus membranaceus, Rehmannia glutinosa, Eucommia ulmoides, Morinda officinalis, Lycium barbarum and Polygonatum sibiricum. Regarding anti-oxidation / aging drugs, including patent CN108310212A-a drug for delaying aging, its preparation method and application, the formula is: 100-200 parts of Codonopsis pilosula, 100-150 parts of Astragalus, 100-150 parts of Cornus officinalis, 100-150 parts of Rehmannia glutinosa, 100-150 parts of Lycium barbarum, 100-150 parts of Mulberry, 100-150 parts of Raspberry, 100-150 parts of Polygonatum multiflorum and 100-150 parts of Polygonatum odoratum, etc. It can be seen that the above-mentioned formulas for treating related diseases either contain more types of drugs or have relatively single efficacy. Summary of the invention

[0005] In order to solve the above technical problems, the present invention provides a traditional Chinese medicine composition for treating kidney yang deficiency and having antioxidant function. The raw materials, measured by mass, include: 2-5 parts of polygonatum, 2-5 parts of raspberry, 2-5 parts of cistanche, 2-5 parts of eucommia male flowers and 1-3 parts of cicada fungus fruiting bodies.

[0006] Preferably, the raw materials, measured by mass, include: 3 parts of polygonatum, 3 parts of rubus idaeus, 3 parts of cistanche deserticola, 3 parts of eucommia male flowers and 1 part of cicada fungus fruiting body.

[0007] Preferably, the Chinese medicine composition is an oral preparation; the oral preparation is granules, powders, pills, tablets or capsules.

[0008] The present invention also provides a preparation method of the traditional Chinese medicine composition, comprising the following steps: decocting and extracting polygonatum, raspberry, cistanche deserticola and eucommia male flowers with water, filtering to obtain an extract; concentrating, drying and grinding the extract to obtain an extract powder; mixing the extract powder with cicada fungus fruiting body powder to obtain the traditional Chinese medicine composition.

[0009] Preferably, the decoction extraction is performed twice, the amount of water added per gram of raw material drug during the first decoction extraction is 9-12 mL, and the amount of water added per gram of raw material drug during the second decoction extraction is 6-10 mL.

[0010] More preferably, during the decoction extraction, the water is heated to boiling each time, and the decoction is maintained at a slight boiling for 1.5-2.5 hours, and the two extracts are combined after filtration.

[0011] Preferably, the drying is carried out by oven drying or freeze drying.

[0012] Preferably, the Chinese medicine composition is stored at 2-8°C in a light-proof environment.

[0013] The present invention also provides application of the traditional Chinese medicine composition or the preparation method in preparing drugs for treating kidney yang deficiency and antioxidant drugs.

[0014] Compared with the prior art, the present invention has the following beneficial effects:

[0015] The Chinese medicine composition of the present invention is composed of a variety of natural medicines, which have the effects of tonifying kidney yang, improving essence and blood, regulating body functions, etc. in traditional Chinese medicine theory, and are compatible with each other and work synergistically. Compared with chemical synthetic drugs, the Chinese medicine composition of the present invention has the characteristics of small side effects and high safety.

[0016] The Chinese medicine composition of the present invention has a relatively concise formula, avoiding the problems of complex ingredients and difficult quality control caused by too many drug combinations. At the same time, the Chinese medicine composition of the present invention not only has the effect of treating kidney yang deficiency, but also has an antioxidant effect. On the basis of improving the symptoms of kidney yang deficiency, it can prevent and treat diseases related to oxidative stress, such as aging-related diseases, cardiovascular diseases, neurodegenerative diseases, etc. through antioxidant pathways, and has more comprehensive effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 The effects of different treatments on the body weight of mice in the kidney-yang deficiency model;

[0018] Figure 2 The effects of different treatments on the body temperature of mice in the drug efficacy model of kidney yang deficiency syndrome;

[0019] Figure 3 The effects of different treatments on the grasping power of mice in the drug efficacy model of kidney yang deficiency syndrome;

[0020] Figure 4 The effects of different treatments on the coagulation time (PT) of mice with kidney-yang deficiency syndrome drug efficacy model;

[0021] Figure 5 To investigate the effects of different treatments on the activity of superoxide dismutase (SOD) in mice with kidney-yang deficiency syndrome.

[0022] Figure 6 To investigate the effects of different treatments on the malondialdehyde (MDA) content in mice with kidney-yang deficiency syndrome.

[0023] Figure 7 The effects of different treatments on the number of spontaneous activities of mice with kidney-yang deficiency syndrome drug efficacy model;

[0024] Figure 8 The effects of different treatments on the body weight of aging model mice;

[0025] Fig. 9 The effects of different treatments on glutathione (GSH) content in aging model mice;

[0026] Fig.10 To investigate the effects of different treatments on the carbonyl content of serum proteins in aging model mice;

[0027] Fig.11 The effects of different treatments on the activity of serum superoxide dismutase (SOD) in aging model mice;

[0028] Fig.12 The effects of different treatments on the malondialdehyde (MDA) content in aging model mice;

[0029] Figure 1-Figure 12 middle, # P<0.05vs normal control group, ## P<0.01vs normal control group; * P<0.05vs model group, ** P<0.01vs model group. DETAILED DESCRIPTION

[0030] The invention provides a traditional Chinese medicine composition for treating kidney yang deficiency and having antioxidant function. The raw materials, measured by weight, include: 2-5 parts of polygonatum, 2-5 parts of rubus idaeus, 2-5 parts of cistanche deserticola, 2-5 parts of eucommia male flowers and 1-3 parts of cicadae contagiosum fruiting bodies; preferably, 3 parts of polygonatum, 3 parts of rubus idaeus, 3 parts of cistanche deserticola, 3 parts of eucommia male flowers and 1 part of cicadae contagiosum fruiting bodies.

[0031] Polygonatum: It is the dried rhizome of Yunnan Polygonatum, Polygonatum or Polygonatum multiflorum of the Liliaceae family. It is sweet and neutral in nature. It enters the spleen, lung and kidney meridians. It has the functions of replenishing qi and nourishing yin, strengthening the spleen, moistening the lungs and benefiting the kidneys. It is used for spleen and stomach qi deficiency, fatigue and weakness, insufficient stomach yin, dry mouth and poor appetite, dry cough due to lung deficiency, cough with blood due to overwork, insufficient essence and blood, soreness of waist and knees, premature graying of hair, internal heat and thirst.

[0032] Raspberry: It is the dried fruit of the Chinese raspberry plant of the Rosaceae family. It is sweet, sour and warm in nature. It enters the liver, kidney and bladder meridians. It has the functions of nourishing the kidney, consolidating essence and reducing urine, nourishing the liver and improving eyesight. It is used for spermatorrhea, nocturnal emission, frequent urination, impotence and premature ejaculation, and blurred vision.

[0033] Cistanche deserticola: The dried fleshy stem with scaly leaves of Cistanche deserticola or Cistanche tubulosa, Orobanchaceae. It is sweet, salty and warm in nature. It enters the kidney and large intestine meridians. It can nourish kidney yang, benefit essence and blood, and moisten the intestines and relieve constipation. It is used for kidney yang deficiency, essence and blood deficiency, impotence and infertility, soreness of waist and knees, weakness of tendons and bones, and constipation caused by dry intestines.

[0034] Eucommia male flowers: The flowers that bloom on the Eucommia male tree. Eucommia male flowers have all the health benefits of the plant Eucommia, and are particularly effective in "strengthening the liver, tonifying the kidneys, promoting bowel movements, promoting sleep, and lowering the three highs."

[0035] Cicada fungus: sweet, cold, non-toxic, enters the lung and liver meridians, and is cool in nature. Modern research shows that cicada fungus has many pharmacological effects, which can enhance the body's immune function, regulate the nervous system, relieve fever and pain, improve kidney function, regulate lipid metabolism, and have anti-fatigue, anti-stress, and anti-tumor effects.

[0036] In the Chinese medicine composition of the present invention, polygonatum sibiricum and cicada fungus fruiting bodies are the main medicines, and raspberry, cistanche deserticola, and eucommia male flower are the ministerial medicines. The main and ministerial medicines are in order, and the compatibility is rigorous, and the treatment principle of warming and tonifying kidney yang, replenishing essence and qi and anti-oxidation is followed together. In the prescription, polygonatum sibiricum, as one of the main medicines, mainly plays an important role in the deficiency of kidney essence and qi caused by kidney yang deficiency, can nourish the innate kidney essence, and can nourish the acquired spleen and stomach qi and blood, through the spleen and kidney double tonification, to provide the body with sufficient qi and blood essence and micro-substances, so as to achieve the purpose of improving the symptoms of kidney yang deficiency; the antioxidant effect of cicada fungus fruiting bodies can counteract the oxidative stress reaction generated by the body in the state of kidney yang deficiency, reduce the damage of free radicals to the body, and with polygonatum sibiricum, one warms and tonifies kidney yang, and the other strengthens the body to resist evil, and treats kidney yang deficiency while taking into account anti-oxidation. Raspberry assists the main medicines Polygonatum sibiricum and Cicadae Concha fruiting bodies to strengthen the kidney-tonifying and sperm-solidifying power. At the same time, its warm nature also helps to warm and nourish the kidney yang, and enhance the overall warming yang effect of the prescription; Cistanche can enhance the warming and nourishing kidney yang effect of the main medicine; Eucommia male flower strengthens the nourishment of the liver and kidney, enhances the body's yang energy, and further improves the therapeutic effect of the prescription on kidney yang deficiency syndrome. In addition, the various medicinal flavors in this prescription assist each other. The cold nature of Cicadae Concha fruiting bodies can restrict the warm and dry nature of other warm medicines (raspberry, Cistanche, Eucommia male flower) to a certain extent, making the whole prescription warm but not dry, avoiding excessive warmth and heat to hurt yin, and playing a supporting role; the nature and taste of each medicine are mild or mutually restrained, which plays a role in reconciling the various medicines, so that the effects of each medicine are synergistically exerted, and the purpose of treating kidney yang deficiency syndrome and exerting comprehensive effects such as anti-oxidation is achieved together.

[0037] In the present invention, the Chinese medicine composition is preferably an oral preparation, and more preferably a granule, powder, pill, tablet or capsule.

[0038] The present invention also provides a method for preparing the Chinese medicine composition, comprising the following steps:

[0039] (1) Weigh polygonatum, rubus idaeus, cistanche deserticola and eucommia male flowers according to mass, add water to decoct and extract, and filter to obtain an extract.

[0040] In the present invention, it is preferred that the decoction extraction is performed twice, and the amount of water added per gram of raw material medicine during the first decoction extraction is 9-12mL, more preferably 10mL, and the amount of water added per gram of raw material medicine during the second decoction extraction is 6-10mL, more preferably 8mL. As an implementable method, 150g of Polygonatum sibiricum, Rubus idaeus, Cistanche deserticola and Eucommia ulmoides male flower are weighed, totaling 600g, the amount of water added for the first decoction is 6000mL, and the amount of water added for the second decoction is 4800mL. It is further preferred that the decoction extraction is heated to boiling water each time, and the decoction is kept at a slight boiling for 1.5-2.5h, more preferably 2h, and the two extracts are combined after filtration.

[0041] (2) Concentrating, drying and grinding the extract in step (1) to obtain a dry extract powder.

[0042] In the present invention, there is no special limitation on the concentration method. As an implementable method, reduced pressure concentration is adopted to concentrate the extract to a suitable volume under the condition of a water bath temperature of 60°C to facilitate subsequent drying. The present invention preferably concentrates the extract to 1 / 4-3 / 4 (mass). As an implementable method, every 100g of the raw material drug is concentrated into 50g of extract.

[0043] In the present invention, drying is preferably performed by oven drying or freeze drying, and freeze drying is more preferably performed. As an implementation method, the concentrated extract is placed in a crystallization dish at a low temperature (-75°C) for pre-freezing, and then freeze drying is performed. The model of the freeze drying instrument is buchi Lyovapor TM L-200, freezing parameters are temperature: -54℃, pressure: 0.2mbar. The extract is freeze-dried to form a powder block, which can be ground into powder for later use.

[0044] (3) Weigh the cicada confusa fruiting body powder by mass, and mix the extract dry powder described in step (2) with the cicada confusa fruiting body powder to obtain the traditional Chinese medicine composition.

[0045] In the present invention, the prepared traditional Chinese medicine composition is preferably stored at 2-8° C. in a light-proof environment.

[0046] The Chinese medicine composition prepared by the present invention is in the form of granules and can be directly orally taken as granules; preferably, the granules are mixed with auxiliary materials and adjuvants, and the granules are processed into powders, pills, tablets or capsules, which are convenient to carry and take. The present invention has no special restrictions on the preparation method of the powders, pills, tablets or capsules, and conventional means in the art can be used.

[0047] The present invention also provides application of the Chinese medicine composition or the preparation method in preparing medicines for treating kidney yang deficiency and antioxidant medicines.

[0048] The above contents of the present invention are further described in detail below through specific implementation methods in the form of embodiments. However, this should not be understood as the scope of the above subject matter of the present invention being limited to the following examples. All technologies realized based on the above contents of the present invention belong to the scope of the present invention.

[0049] Example 1

[0050] A Chinese medicine composition for treating kidney yang deficiency and having antioxidant function, the preparation method of which is as follows:

[0051] (1) Weigh 3 parts of polygonatum, 3 parts of raspberry, 3 parts of cistanche, and 3 parts of eucommia male flower by mass, add water and decoct for extraction. The amount of water added for the first decoction is 10 mL per gram of raw material drug, and the amount of water added for the second decoction is 8 mL per gram of raw material drug. Heat the water to boiling for each decoction, keep it at a slight boiling for 2 hours, and filter and combine the two extracts;

[0052] (2) concentrating the extract of step (1) under reduced pressure to 1 / 2 of its mass, freeze-drying, and grinding to obtain freeze-dried powder;

[0053] (3) Weigh 1 portion of cicada fungus fruiting body powder and mix it evenly with the freeze-dried powder of step (2) to obtain a traditional Chinese medicine composition.

[0054] Example 2

[0055] A Chinese medicine composition for treating kidney yang deficiency and having antioxidant function, the preparation method of which is as follows:

[0056] (1) Weigh 5 parts of polygonatum, 5 parts of raspberry, 5 parts of cistanche deserticola, and 5 parts of eucommia male flower by weight, add water and decoct for extraction. The amount of water added for the first decoction is 10 mL per gram of raw material drug, and the amount of water added for the second decoction is 8 mL per gram of raw material drug. Heat the water to boiling for each decoction and keep it at a slight boiling for 2.5 hours. Filter and combine the two extracts;

[0057] (2) concentrating the extract obtained in step (1) to 3 / 4 of its mass under reduced pressure, drying it, and grinding it to obtain a dried powder;

[0058] (3) Weigh 3 parts of cicada fungus fruiting body powder and mix it evenly with the dried powder of step (2) to obtain a Chinese medicine composition.

[0059] Example 3

[0060] A Chinese medicine composition for treating kidney yang deficiency and having antioxidant function, the preparation method of which is as follows:

[0061] (1) Weigh 4 parts of polygonatum, 2 parts of raspberry, 4 parts of cistanche, and 5 parts of eucommia male flower by weight, add water and decoct for extraction. The amount of water added for the first decoction is 10 mL per gram of raw material drug, and the amount of water added for the second decoction is 8 mL per gram of raw material drug. Heat the water to boiling for each decoction and keep it at a slight boiling for 1.5 hours. Filter and combine the two extracts;

[0062] (2) concentrating the extract of step (1) under reduced pressure to 1 / 4 of its mass, freeze-drying, and grinding to obtain freeze-dried powder;

[0063] (3) Weigh 2 parts of cicada fungus fruiting body powder and mix it evenly with the freeze-dried powder of step (2) to obtain a traditional Chinese medicine composition.

[0064] Example 4

[0065] A Chinese medicine composition for treating kidney yang deficiency and having antioxidant function, the preparation method of which is as follows:

[0066] (1) Weigh 5 parts of polygonatum, 4 parts of raspberry, 2 parts of cistanche, and 4 parts of eucommia male flower by weight, add water and decoct for extraction. The amount of water added for the first decoction is 10 mL per gram of raw material drug, and the amount of water added for the second decoction is 8 mL per gram of raw material drug. Heat the water to boiling for each decoction and keep it at a slight boiling for 2 hours. Filter and combine the two extracts;

[0067] (2) concentrating the extract of step (1) under reduced pressure to 1 / 2 of its mass, freeze-drying, and grinding to obtain freeze-dried powder;

[0068] (3) Weigh 2 parts of cicada fungus fruiting body powder and mix it evenly with the freeze-dried powder of step (2) to obtain a traditional Chinese medicine composition.

[0069] Example 5

[0070] Experimental study on the efficacy of the Chinese medicine composition (Cicada Flower and Polygonatum Sinensis Paste) on mice with kidney yang deficiency syndrome

[0071] 1. Instruments and Materials

[0072] 1.1 Instrument

[0073] Thermometer (Deli probe thermometer); ELISA reader (Thermo Scientific Varioskan Flash); rat and mouse grip strength tester (Shanghai Xinruan XR501); spontaneous activity recorder (Yiyan Technology spontaneous activity recorder YLS-1A).

[0074] 1.2 Materials

[0075] 1.2.1 Test products

[0076] Cicada fungus and Polygonatum sibiricum paste. Storage conditions: 2-8℃, avoid light.

[0077] Polygonatum odoratum paste. Storage conditions: 2-8℃, avoid light.

[0078] Hydrocortisone acetate tablets, brand: Xinyi; specification: 20mg.

[0079] MDAAssayKit, brand: abcam; specification: 100test.

[0080] SOD test kit, brand: Nanjing Jiancheng; specification: 96test.

[0081] 1.2.2 Experimental animals

[0082] 40 SPF-grade ICR male mice, weighing 18-22 g, were purchased from Shanghai Shengchang Biotechnology Co., Ltd.; certificate number: 20210002007678.

[0083] 2. Test methods

[0084] 2.1 Animal husbandry

[0085] Animals were fed with standard sterilized mouse feed, and drinking water was provided by drinking bottles. Animals were free to drink water. Ten animals were kept in each cage. The room temperature was set at 20℃-22℃, humidity was 40%-70%, and the light was alternating between light and dark for 12 hours. The bedding was changed at least twice a week, and the feeding box was changed at the same time. The feeding box was changed at any time when there was an abnormal situation. The drinking water bottle and bottle stopper were replaced and sterilized every day, and the cage rack was disinfected once every two weeks. All cages were sterilized by high pressure after cleaning.

[0086] 2.2 Animal grouping

[0087] Sixty mice were randomly divided into six groups according to their body weight as shown in Table 1.

[0088] Table 1 Animal grouping table of mouse kidney yang deficiency model test

[0089] Animal Grouping quantity Normal control group 10 Model Group 10 High-dose 6.76g herbal medicine / kg group of Cicadae and Polygonatum odoratum paste 10 Low-dose 1.69g crude drug / kg group of Cicadae Polygonatum odoratum paste 10 Huangjing ointment high dose 6.24g crude drug / kg group 10 Huangjing paste low dose 1.56g crude drug / kg group 10

[0090] 2.3 Drug preparation and administration route

[0091] Preparation of cicada flower and polygonatum paste: 150g each of polygonatum, raspberry, cistanche deserticola, and eucommia male flower are taken, deionized water is added, and the extraction is heated and decocted twice. The amount of water added for the first decoction is 6000mL, and the amount of water added for the second decoction is 4800mL. Each decoction is heated to boiling water and kept at a slight boiling for 2h; filter, combine the two extracts, concentrate under reduced pressure to an appropriate volume, and freeze-dry (the model of the freeze-drying instrument is buchi Lyovapor TML-200, freezing parameters are temperature: -54℃, pressure: 0.2mbar), and freeze-dried powder is obtained. 600g of raw material is added to obtain 323.42g of freeze-dried powder, and the yield of freeze-dried powder is 53.90%. 50g of cicada fungus fruiting body powder is taken and mixed with the above freeze-dried powder to obtain cicada fungus and polygonatum paste. The total mass is 373.42g / 650g raw material, which is equivalent to a yield of 57.45%.

[0092] Low-dose 1.69 g crude drug / kg group of Cicadae Concha and Polygonatum sibiricum paste: 0.970 g of Cicadae Concha and Polygonatum sibiricum paste was measured and dissolved in 20 mL of pure water to prepare 0.0485 g / mL Cicadae Concha and Polygonatum sibiricum paste liquid for administration.

[0093] High-dose 6.76 g crude drug / kg group of Cicadae Concha and Polygonatum sibiricum paste: 3.880 g of Cicadae Concha and Polygonatum sibiricum paste was measured and dissolved in 20 mL of pure water to prepare 0.1940 g / mL Cicadae Concha and Polygonatum sibiricum paste liquid for administration.

[0094] Preparation of Polygonatum sibiricum Paste: The preparation method is the same as that of Cicadella Concha Polygonatum sibiricum Paste, except that Cicadella Concha fruiting body powder is not added subsequently. The yield of the freeze-dried powder is 44.08%.

[0095] Polygonatum sibiricum low-dose 1.56 g crude drug / kg group: 0.688 g of Polygonatum sibiricum was measured and dissolved in 20 mL of pure water to prepare 0.0344 g / mL Polygonatum sibiricum liquid for administration.

[0096] Polygonatum sibiricum high-dose 6.24 g crude drug / kg group: 2.752 g of Polygonatum sibiricum was measured and dissolved in 20 mL of pure water to prepare 0.1380 g / mL Polygonatum sibiricum liquid for administration.

[0097] Hydrocortisone modeling agent: Dosage: 50 mg / kg.

[0098] Route of administration: intragastric administration.

[0099] 2.4 Model replication and drug administration process

[0100] After adaptive feeding, the mice were divided into groups and given drugs for 17 days. On the 18th, 19th, 20th and 21st days, 50 mg / kg of hydrocortisone was gavaged daily 1 hour after drug administration. One hour after the last drug administration on the 22nd day, the physical signs of the mice were observed (the weight of the mice was weighed with a scale; the rectal temperature of the mice was measured with a thermometer; the number of spontaneous activities of the mice within 5 minutes was detected with an autonomic activity meter; the gripping strength of the forelimbs of the mice was detected with an animal gripping strength meter; after the test, the eyeballs were removed to collect blood for the determination of coagulation time, and the blood was collected and centrifuged to obtain serum, and the serum superoxide dismutase (SOD) and malondialdehyde (MDA) were detected with a kit.

[0101] 2.5 Statistical methods

[0102] For the count data, this example used a two-sided t-test between groups for statistical comparison, and P<0.05 was considered to be statistically significant.

[0103] 3. Test results

[0104] 3.1 Effects of different treatments on the body weight of mice in the kidney-yang deficiency model

[0105] As shown in Table 2 and Figure 1 As shown in the figure, compared with the normal control group, the body weight of the mice in the model group decreased after modeling, with a statistical difference (P<0.05). Compared with the model group, the body weight of the 6.76g / kg group of Cicadae Concha and Polygonatum Ointment decreased, with a statistical difference (P<0.05); there was no statistical difference in the body weight of the mice in the 1.69g / kg group of Cicadae Concha and Polygonatum Ointment, the 6.24g / kg group of Polygonatum Ointment and the 1.56g / kg group compared with the model group (P>0.05).

[0106] Table 2 Effects of different treatments on body weight of mice in kidney yang deficiency model

[0107] Group dose Weight (g) Normal control group / 37.0±3.5 Model Group / <![CDATA[34.1±1.9 # ]]> High-dose group of cicada flower and polygonatum ointment 6.76g crude drug / kg <![CDATA[31.6±2.8 * <!-- 6 -->]]> Low-dose group of cicada flower and polygonatum ointment 1.69g crude drug / kg 33.0±3.3 Huangjing ointment high dose group 6.24g crude drug / kg 33.8±2.7 Huangjing ointment low dose group 1.56g crude drug / kg 32.0±3.6

[0108] In the table, # P<0.05vs normal control group, ## P<0.01vs normal control group; * P<0.05vs model group, ** P<0.01vs model group. Same below.

[0109] 3.2 Effects of different treatments on body temperature of mice in the kidney-yang deficiency model

[0110] As shown in Table 3 and Figure 2 As shown in the figure, compared with the normal control group, the body temperature of the mice in the model group decreased after modeling, with a statistically significant difference (P<0.01). Compared with the model group, the body temperature of the Cordyceps sinensis and Polygonatum sibiricum paste 6.76g / kg group, Cordyceps sinensis and Polygonatum sibiricum paste 1.69g / kg group, Polygonatum sibiricum paste 6.24g / kg group and Polygonatum sibiricum paste 1.56g / kg group all increased, with a statistically significant difference (P<0.05 or P<0.01).

[0111] Table 3 Effects of different treatments on body temperature of mice in kidney yang deficiency model

[0112] Group dose Body temperature(℃) Normal control group / 38.0±0.3 Model Group / <![CDATA[36.9±0.5 ## ]]> High-dose group of cicada flower and polygonatum ointment 6.76g crude drug / kg <![CDATA[37.7±0.4 ** ]]> Low-dose group of cicada flower and polygonatum ointment 1.69g crude drug / kg <![CDATA[37.6±0.3 ** ]]> Huangjing ointment high dose group 6.24g crude drug / kg <![CDATA[37.8±0.4 * ]]> Huangjing ointment low dose group 1.56g crude drug / kg <![CDATA[37.3±0.2 ** ]]>

[0113] 3.3 Effects of different treatments on the grip strength of mice in the kidney-yang deficiency model

[0114] As shown in Table 4 and Figure 3As shown in the figure, compared with the normal control group, the grasping force of the mice in the model group decreased after modeling, with a statistical difference (P<0.01). Compared with the model group, the grasping force of the mice in the 6.76g / kg group of Cicadae Concha and Polygonatum Sinensis Paste increased, with a statistical difference (P<0.05); compared with the model group, there was no difference in the grasping force of the mice in the 1.69g / kg group of Cicadae Concha and Polygonatum Sinensis Paste, the 6.24g / kg group of Polygonatum Sinensis Paste, and the 1.56g / kg group of Polygonatum Sinensis Paste.

[0115] Table 4 Effects of different treatments on the gripping strength of mice with kidney yang deficiency syndrome drug efficacy model

[0116] Group dose Grip force(g) Normal control group / 377.2±42.1 Model Group / <![CDATA[257.4±49.8 ## ]]> High-dose group of cicada flower and polygonatum ointment 6.76g crude drug / kg <![CDATA[313.3±47.4 * ]]> Low-dose group of cicada flower and polygonatum ointment 1.69g crude drug / kg 258.0±57.4 Huangjing ointment high dose group 6.24g crude drug / kg 297.1±83 Huangjing ointment low dose group 1.56g crude drug / kg 268.9±83.5

[0117] 3.4 Effects of different treatments on coagulation time in mice with kidney-yang deficiency syndrome

[0118] As shown in Table 5 and Figure 4 As shown in the figure, compared with the normal control group, the coagulation time of the mice in the model group was prolonged after modeling, with a statistically significant difference (P<0.01). Compared with the model group, the coagulation time of the mice in the 6.76g / kg group of Cicadae and Polygonatum sibiricum and the 6.24g / kg group of Polygonatum sibiricum was shortened, but there was no statistical difference (P>0.05); compared with the model group, the coagulation time of the mice in the 1.69g / kg group of Cicadae and Polygonatum sibiricum and the 1.56g / kg group of Polygonatum sibiricum was not different.

[0119] Table 5 Effects of different treatments on coagulation time in mice with kidney yang deficiency syndrome

[0120] Group dose Coagulation time (s) Normal control group / 38.7±8.6 Model Group / <![CDATA[60.0±14.1 ## ]]> High-dose group of cicada flower and polygonatum ointment 6.76g crude drug / kg 52.1±17.4 Low-dose group of cicada flower and polygonatum ointment 1.69g crude drug / kg 57.0±16.1 Huangjing ointment high dose group 6.24g crude drug / kg 51.0±11.1 Huangjing ointment low dose group 1.56g crude drug / kg 56.3±18.8

[0121] 3.5 Effects of different treatments on superoxide dismutase (SOD) activity in mice with kidney-yang deficiency syndrome

[0122] As shown in Table 6 and Figure 5 As shown in the figure, compared with the normal control group, the serum SOD activity of the model group mice decreased after modeling, with statistical difference (P<0.01). Compared with the model group, the serum SOD activity of the mice in the 6.76g / kg group of Cicadae Concha Polygonatum Ointment, the 1.69g / kg group of Cicadae Concha Polygonatum Ointment, and the 6.24g / kg group of Polygonatum Ointment increased, with statistical difference (P<0.05 or P<0.01). Compared with the model group, there was no difference in the serum SOD activity of the mice in the 1.56g / kg group of Polygonatum Ointment.

[0123] Table 6 Effects of different treatments on superoxide dismutase (SOD) activity in mice with kidney yang deficiency syndrome

[0124]

[0125]

[0126] 3.6 Effects of different treatments on serum malondialdehyde (MDA) content in mice with kidney-yang deficiency syndrome

[0127] As shown in Table 7 and Figure 6 As shown in the figure, compared with the normal control group, the serum MDA content of the mice in the model group increased after modeling, with a statistically significant difference (P<0.01). Compared with the model group, the serum MDA content of the mice in the 6.76g / kg group of Cicadae Concha and Polygonatum Sinensis Ointment decreased, with a statistically significant difference (P<0.05); compared with the model group, the serum MDA content of the mice in the 1.69g / kg group of Cicadae Concha and Polygonatum Sinensis Ointment did not differ. There was no difference in the serum MDA content of the mice in the 1.69g / kg group of Cicadae Concha and Polygonatum Sinensis Ointment, the 6.24g / kg group of Polygonatum Sinensis Ointment, and the 1.56g / kg group of Polygonatum Sinensis Ointment.

[0128] Table 7 Effects of different treatments on serum malondialdehyde (MDA) content in mice with kidney yang deficiency syndrome

[0129] Group dose MDA (umol / L) Normal control group / 11.931±2.887 Model Group / <![CDATA[15.974±2.536 ## ]]> High-dose group of cicada flower and polygonatum ointment 6.76g crude drug / kg <![CDATA[11.540±1.777 * ]]> Low-dose group of cicada flower and polygonatum ointment 1.69g crude drug / kg 12.250±1.718 Huangjing ointment high dose group 6.24g crude drug / kg 15.499±2.451 Huangjing ointment low dose group 1.56g crude drug / kg 13.222±2.802

[0130] 3.7 Effects of different treatments on the number of spontaneous activities in mice with kidney-yang deficiency syndrome

[0131] As shown in Table 8 and Figure 7 As shown in the figure, compared with the normal control group, the spontaneous activity of mice in the model group increased after modeling, with statistical difference (P<0.01). Compared with the model group, the spontaneous activity of mice in the 6.76g / kg group of Cicadae Concha Polygonatum Ointment, the 1.69g / kg group of Cicadae Concha Polygonatum Ointment, the 6.24g / kg group of Polygonatum Ointment and the 1.56g / kg group of Polygonatum Ointment decreased, with statistical difference (P<0.05 or P<0.01).

[0132] Table 8 Effects of different treatments on the number of spontaneous activities in mice with kidney yang deficiency syndrome

[0133] Group dose Number of independent activities Normal control group / 60.0±10.9 Model Group / <![CDATA[82.4±18.0 ## ]]> High-dose group of cicada flower and polygonatum ointment 6.76g crude drug / kg <![CDATA[53.6±20.5 ** ]]> Low-dose group of cicada flower and polygonatum ointment 1.69g crude drug / kg <![CDATA[49.5±17.1 ** ]]> Huangjing ointment high dose group 6.24g crude drug / kg <![CDATA[62.3±15.5 ** ]]> Huangjing ointment low dose group 1.56g crude drug / kg <![CDATA[57.7±20.2 * ]]>

[0134] Based on the above test results, it was found that the Chinese medicine composition of the present invention (Cicada Cone and Polygonatum sibiricum Paste) caused the weight loss of mice, with statistical significance (P<0.05); it was able to increase the body temperature of mice with kidney yang deficiency model, enhance the grip strength of mice with kidney yang deficiency model, increase the serum SOD activity of mice with kidney yang deficiency model, reduce the serum MDA content of mice with kidney yang deficiency model, and reduce the number of autonomous activities of mice, but the effect on the coagulation time of mice with kidney yang deficiency model was not obvious.

[0135] Example 6

[0136] Experimental study on the efficacy of the Chinese medicine composition (Cicada Flower and Polygonatum odoratum paste) on mice aging model

[0137] 1. Instruments and Materials

[0138] 1.1 Instrument

[0139] Microplate reader (Thermo Scientific Varioskan Flash).

[0140] 1.2 Materials

[0141] 1.2.1 Test products

[0142] Cicada fungus and Polygonatum sibiricum paste. Storage conditions: 2-8℃, avoid light.

[0143] Polygonatum odoratum paste. Storage conditions: 2-8℃, avoid light.

[0144] D-galactose, brand: Macklin; specification: 500g.

[0145] MDAAssayKit, brand: abcam; specification: 100test.

[0146] SOD test kit, brand: Nanjing Jiancheng; specification: 96test;

[0147] GSH test kit, brand: Nanjing Jiancheng; specification: 96test;

[0148] Protein Carbony Content Assay Kit, brand: Sigma; specification: 96 tests.

[0149] 1.2.2 Experimental animals

[0150] 40 SPF-grade ICR male mice, weighing 18-22 g, were purchased from Shanghai Shengchang Biotechnology Co., Ltd.; certificate number: 20210002007726.

[0151] 2. Test methods

[0152] 2.1 Animal husbandry

[0153] Animals were fed with standard sterilized mouse feed, and drinking water was provided by drinking bottles. Animals were free to drink water. Ten animals were kept in each cage. The room temperature was set at 20℃-22℃, humidity was 40%-70%, and the light was alternating between light and dark for 12 hours. The bedding was changed at least twice a week, and the feeding box was changed at the same time. The feeding box was changed at any time when there was an abnormal situation. The drinking water bottle and bottle stopper were replaced and sterilized every day, and the cage rack was disinfected once every two weeks. All cages were sterilized by high pressure after cleaning.

[0154] 2.2 Animal grouping

[0155] 60 mice were randomly divided into 6 groups according to their body weight as shown in Table 9.

[0156] Table 9 Grouping of animals in the mouse aging model test

[0157] Animal Grouping quantity Normal control group 10 Model Group 10 High-dose 6.76g herbal medicine / kg group of Cicadae and Polygonatum odoratum paste 10 Low-dose 1.69g crude drug / kg group of Cicadae Polygonatum odoratum paste 10 Huangjing ointment high dose 6.24g crude drug / kg group 10 Huangjing paste low dose 1.56g crude drug / kg group 10

[0158] 2.3 Drug preparation and administration route

[0159] The preparations of Cicadae Concha and Polygonatum Sinensis Paste ... are the same as those in Example 5.

[0160] Modeling agent: 5% D-galactose; Preparation: 5 g of D-galactose was added to physiological saline and dissolved to 100 mL.

[0161] Route of administration: intragastric administration.

[0162] 2.4 Model replication and drug administration process

[0163] After adaptive feeding, the mice were divided into groups and given drugs for 21 days. After the drugs were given, 0.1 mL of 5% D-galactose was injected subcutaneously. One hour after the last drug administration on the 22nd day, the physical signs of the mice were observed. After the test, the eyeballs were removed to collect blood. After the blood was collected, it was centrifuged to obtain serum, and malondialdehyde, superoxide dismutase, protein carbonyl, and reduced glutathione were detected using a kit.

[0164] 2.5 Statistical methods

[0165] For the count data, this example used a two-sided t-test between groups for statistical comparison, and P<0.05 was considered to be statistically significant.

[0166] 3. Experimental results

[0167] 3.1 Effects of different treatments on the body weight of aging model mice

[0168] As shown in Table 10 and Figure 8 As shown in the figure, compared with the normal control group, the body weight of mice in the model group decreased after modeling, but there was no statistical difference (P>0.05). Compared with the model group, the body weight of mice in each group was not significantly different from that of the model group (P>0.05).

[0169] Table 10 Effects of different treatments on body weight of aging model mice

[0170]

[0171]

[0172] 3.2 Effects of different treatments on glutathione (GSH) content in aging model mice

[0173] As shown in Table 11 and Fig. 9As shown in the figure, compared with the normal control group, the GSH of the mice in the model group decreased after modeling, with a statistical difference (P<0.01). Compared with the model group, the GSH of the mice in the 6.76g / kg group of Cicadae and Polygonatum sibiricum Paste and the 6.24g / kg group of Polygonatum sibiricum Paste increased, with a statistical difference (P<0.01). Compared with the model group, the GSH of the mice in the 1.69g / kg group of Cicadae and Polygonatum sibiricum Paste and the 1.56g / kg group of Polygonatum sibiricum Paste had no statistical difference compared with the model group (P>0.05).

[0174] Table 11 Effects of different treatments on glutathione (GSH) content in aging model mice

[0175] Group dose GSH (nmol / L) Normal control group / 70.56±23.65 Model Group / <![CDATA[27.07±13.63 ## ]]> High-dose group of cicada flower and polygonatum ointment 6.76g crude drug / kg <![CDATA[69.83±29.46 ** ]]> Low-dose group of cicada flower and polygonatum ointment 1.69g crude drug / kg 38.00±16.88 Huangjing ointment high dose group 6.24g crude drug / kg <![CDATA[65.93±26.59 ** ]]> Huangjing ointment low dose group 1.56g crude drug / kg 38.57±20.78

[0176] 3.3 Effects of different treatments on protein carbonyl content in aging model mice

[0177] As shown in Table 12 and Fig.10 As shown in the figure, compared with the normal control group, the serum protein carbonyl of the model group mice increased after modeling, with a statistically significant difference (P<0.01). Compared with the model group, the serum protein carbonyl of the mice in each drug administration group decreased, with a statistically significant difference (P<0.01).

[0178] Table 12 Effects of different treatments on protein carbonyl content in aging model mice

[0179] Group dose Protein carbonyl (nmol / mgP) Normal control group / 29.19±5.19 Model Group / <![CDATA[57.42±4.47 ## ]]> High-dose group of cicada flower and polygonatum ointment 6.76g crude drug / kg <![CDATA[35.28±11.97 ** ]]> Low-dose group of cicada flower and polygonatum ointment 1.69g crude drug / kg <![CDATA[26.48±3.77 ** ]]> Huangjing ointment high dose group 6.24g crude drug / kg <![CDATA[40.68±2.91 ** ]]> Huangjing ointment low dose group 1.56g crude drug / kg <![CDATA[36.49±3.22 ** ]]>

[0180] 3.4 Effects of different treatments on superoxide dismutase (SOD) activity in aging model mice

[0181] As shown in Table 13 and Fig.11 As shown in the figure, compared with the normal control group, the serum SOD activity of the model group mice decreased after modeling, with a statistically significant difference (P<0.01). Compared with the model group, the serum SOD activity of the mice in the 6.76g / kg group of Cicadae Polygonatum Ointment and the 6.24g / kg group of Polygonatum Ointment increased, with a statistically significant difference (P<0.05); compared with the model group, there was no difference in the serum SOD activity of the mice in the 1.69g / kg group of Cicadae Polygonatum Ointment and the 1.56g / kg group of Polygonatum Ointment.

[0182] Table 13 Effects of different treatments on superoxide dismutase (SOD) activity in aging model mice

[0183] Group dose SOD activity (U / mL) Normal control group / 65.92±2.85 Model Group / <![CDATA[55.39±5.95 ## ]]> High-dose group of cicada flower and polygonatum ointment 6.76g crude drug / kg <![CDATA[61.23±2.50 * ]]> Low-dose group of cicada flower and polygonatum ointment 1.69g crude drug / kg 57.09±3.73 Huangjing ointment high dose group 6.24g crude drug / kg <![CDATA[62.03±4.38 * ]]> Huangjing ointment low dose group 1.56g crude drug / kg 59.63±5.22

[0184] 3.5 Effects of different treatments on serum malondialdehyde (MDA) content in aging model mice

[0185] As shown in Table 14 and Fig.12As shown in the figure, compared with the normal control group, the serum MDA content of the mice in the model group increased after modeling, with statistical difference (P<0.05). Compared with the model group, the serum MDA content of the mice in the 6.76g / kg group of Cicadae Polygonatum Ointment and the 1.56g / kg group of Polygonatum Ointment decreased, with statistical difference (P<0.01 or P<0.05); compared with the model group, the serum MDA content of the mice in the 1.69g / kg group of Cicadae Polygonatum Ointment and the 6.24g / kg group of Polygonatum Ointment did not differ.

[0186] Table 14 Effects of different treatments on serum malondialdehyde (MDA) content in aging model mice

[0187] Group dose MDA (umol / L) Normal control group / 16.64±3.90 Model Group / <![CDATA[19.74±2.41 # ]]> High-dose group of cicada flower and polygonatum ointment 6.76g crude drug / kg <![CDATA[16.39±3.53 * ]]> Low-dose group of cicada flower and polygonatum ointment 1.69g crude drug / kg 18.86±3.65 Huangjing ointment high dose group 6.24g crude drug / kg 18.58±2.65 Huangjing ointment low dose group 1.56g crude drug / kg <![CDATA[16.86±1.81 ** ]]>

[0188] Excess free radicals may cause damage to a variety of biomacromolecules, and it is speculated that it will have an important impact on gene expression and regulatory systems, and the outcome is to change the differentiation state of cells and lead to aging changes. The aging reaction induced by D-galactose will also show this feature. Based on the experimental results of this embodiment, it is found that the Chinese medicine composition of the present invention (Cicada Flower and Polygonatum Cream) has a slight effect on the body weight of D-galactose aging model mice, but there is no statistical difference; it can increase the GSH of D-galactose aging mice, reduce the serum protein carbonyl of D-galactose aging model mice; increase the serum SOD activity of D-galactose aging model mice; reduce the content of serum MDA in D-galactose aging model mice, and has an antioxidant effect.

[0189] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A Chinese medicine composition for treating kidney yang deficiency and having antioxidant function, characterized in that: The raw materials calculated by weight include: 2-5 parts of polygonatum, 2-5 parts of raspberry, 2-5 parts of cistanche, 2-5 parts of eucommia male flowers and 1-3 parts of cicada fungus fruiting bodies.

2. The Chinese medicine composition according to claim 1, characterized in that: The raw materials measured by mass include: 3 parts of polygonatum, 3 parts of rubus idaeus, 3 parts of cistanche deserticola, 3 parts of eucommia male flowers and 1 part of cicada fungus fruiting body.

3. The Chinese medicine composition according to claim 1 or 2, characterized in that: The Chinese medicine composition is an oral preparation; the oral preparation is granules, powders, pills, tablets or capsules.

4. A method for preparing the Chinese medicine composition according to claim 1 or 2, characterized in that: The following steps are involved: Polygonatum sibiricum, raspberry, Cistanche deserticola and Eucommia male flowers are decocted with water for extraction, and an extract is obtained by filtering; the extract is concentrated, dried and ground to obtain an extract powder; the extract powder is mixed with Cicadae confusa fruiting body powder to obtain the traditional Chinese medicine composition.

5. The preparation method according to claim 4, characterized in that: The decoction extraction is performed twice, 9-12 mL of water is added per gram of the raw material drug during the first decoction extraction, and 6-10 mL of water is added per gram of the raw material drug during the second decoction extraction.

6. The preparation method according to claim 5, characterized in that: During the decoction extraction, heat the water to boiling each time, keep it at a slight boiling point for 1.5-2.5 hours, and combine the two extracts after filtering.

7. The preparation method according to claim 4, characterized in that: The drying is carried out by oven drying or freeze drying.

8. The preparation method according to claim 4, characterized in that: The Chinese medicine composition is stored at 2-8° C. in a light-proof environment.

9. Use of the Chinese medicine composition according to any one of claims 1 to 3 or the preparation method according to any one of claims 4 to 8 in preparing a medicine for treating kidney-yang deficiency.

10. Use of the Chinese medicine composition according to any one of claims 1 to 3 or the preparation method according to any one of claims 4 to 8 in the preparation of antioxidant drugs.

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