A preparation method and application of Huajuhong and Huajuhongtai for relieving cough, reducing phlegm, protecting lungs, enhancing efficacy and removing bitterness

Abstract

The present invention belongs to the technical field of traditional Chinese medicine, and specifically relates to a preparation method and application of Huajuhong and Huajuhong embryo for relieving cough, reducing phlegm, protecting lungs, enhancing efficacy and removing bitterness. The preparation method of the present invention is: take the raw materials (Huajuhong, Huajuhong embryo) and mix them with sodium bicarbonate aqueous solution or sodium carbonate aqueous solution, warm them at 50-90°C, and then dry them to obtain the prepared products. Experimental results show that the preparation method of Huajuhong and Huajuhong embryo of the present invention significantly changes the content of their ingredients, and the efficacy of the prepared prepared products is significantly improved, including a significant increase in the efficacy and activity of relieving cough, reducing phlegm and protecting lungs; moreover, the content of the main bitter substance naringin is significantly reduced after preparation, and the bitterness in the raw materials can be effectively removed, which is very beneficial for oral drinking by patients with cough, phlegm and lung damage.

Description

Technical Field

[0001] The present invention belongs to the technical field of traditional Chinese medicine, and more specifically relates to a preparation method and application of Huajuhong and Huajuhongtai for relieving cough, reducing phlegm, protecting lungs, enhancing efficacy and removing bitterness. Background Art

[0002] Exocarpium Citri Grandis, listed in the Chinese Pharmacopoeia, is the immature or nearly mature dried outer peel of Citrus grandis 'Tomentosa' or Citrus grandis (L.) Osbeck, both Rutaceae plants. The former is commonly known as "hairy orange peel," while the latter is commonly known as "light seven claws" or "light five claws." The fruit is harvested in the summer when it is immature, briefly blanched in boiling water, and then cut into 5 or 7 segments. The pulp and part of the mesocarp are removed, pressed into shape, and dried. Exocarp embryos are the dried young fruit of Citrus grandis 'Tomentosa', a Rutaceae plant, listed in the Guangdong Provincial Medicinal Materials Standard. The fruit is harvested in late spring and early summer when it is immature, withered, and dried; or withered and then pressed into cylindrical shapes and dried; or withered and then cut into two or four segments longitudinally and dried; or sliced, withered, and then dried. The surface of the Huajuhong tea is covered with tiny white hairs, and there is a distinct fragrance beneath the fine hairs. Due to long-term aging and unique processing technology, it is brown in color and has a firm texture.

[0003] Both Huajuhong (Huajuhong) and Huajuhongtai (Huajuhong) are used as traditional Chinese medicines. They have the properties of regulating qi, relieving fullness, and dispelling dampness and resolving phlegm. They are used for coughs with excessive phlegm, food stagnation, alcohol poisoning, vomiting, nausea, and abdominal distension. Huajuhong and Huajuhongtai are specialty products of Huazhou City, Maoming City, Guangdong Province, and are designated as China's National Geographical Indication Products.

[0004] Processing is a hallmark of Traditional Chinese Medicine (TCM) and a key method for enhancing efficacy and improving flavor. Currently, the processing techniques for Huajuhong (Huajuhong) include stir-frying, salting, honey roasting, earthenware processing, ginger processing, and alum processing. Naringin is one of the main active ingredients in Huajuhong (Huajuhong) and Huajuhong embryo, and it is also the primary component responsible for its bitterness. Therefore, the challenge in preparing Huajuhong and Huajuhong embryo lies in achieving a bitter effect while maintaining or even enhancing efficacy. Summary of the Invention

[0005] The technical problem to be solved by the present invention is to overcome the defects and shortcomings of the existing processing technology, and provide a method for processing citrus peel and citrus peel embryo to enhance the effect and remove the bitterness.

[0006] The purpose of the present invention is to provide the application of the synergistic and / or bitter-removing processing method of the present invention in the preparation of synergistic and / or bitter-removing Huajuhong or Huajuhongtai processed products.

[0007] The purpose of the present invention is to provide a processed product of Huajuhong or Huajuhongtai prepared by the synergistic and bitter-removing processing method of the present invention.

[0008] Another object of the present invention is to provide the use of the Huajuhong or Huajuhongtai processed product in the preparation of medicaments for relieving cough, resolving phlegm and / or protecting the lungs.

[0009] Another object of the present invention is to provide a medicine for relieving cough, resolving phlegm and / or protecting the lungs, which contains the processed product of Huajuhong or Huajuhongtai described in the present invention.

[0010] Another object of the present invention is to provide a medicine for relieving cough, resolving phlegm and / or protecting the lungs, which contains the processed product of Huajuhong or Huajuhongtai described in the present invention, as well as other Chinese medicines with medicinal and edible properties other than Huajuhong or Huajuhongtai.

[0011] The above-mentioned purpose of the present invention is achieved through the following technical solutions:

[0012] A method for preparing Huajuhong and Huajuhongtai to enhance the bitterness and dispel the bitterness of Huajuhong is prepared according to the following steps:

[0013] Take the raw material of Huajuhong or Huajuhong embryo, mix it with a sodium bicarbonate aqueous solution or a sodium carbonate aqueous solution with a mass volume concentration of 0.5-10%, soak it at 50-90° C. for 6-72 hours, and then dry it to obtain a processed product.

[0014] Experiments conducted by the present invention have revealed significant changes in the contents of various substances in the processed product obtained using the synergistic and bitter-removing processing method described herein. These changes include a relative decrease in the dissolution of naringin, the major bitter component, and a relative increase in the contents of flavonoids such as naringenin-7-O-glucoside, naringenin, and naringin dihydrochalcone. Furthermore, efficacy experiments have shown that the cough-relieving, expectorant, and lung-protecting activities of Huajuhong processed using the synergistic and bitter-removing processing method described herein are significantly higher than those of Huajuhong processed without processing. This indicates that the synergistic and bitter-removing processing method described herein can enhance the efficacy of Huajuhong while simultaneously reducing its bitterness.

[0015] The Huajuhong described in the present invention is the immature or nearly mature dried outer peel of the Rutaceae plant Huazhou pomelo Citrus grandis'Tomentosa' or Citrus grandis (L.) Osbeck, which has not been otherwise processed; the Huajuhong embryo described in the present invention is the dried young fruit of the Rutaceae plant Huazhou pomelo Citrus grandis'Tomentosa', which has not been otherwise processed.

[0016] Preferably, in the preparation method, the mass volume concentration of the sodium bicarbonate aqueous solution or the sodium carbonate aqueous solution is 0.5-5%.

[0017] Preferably, in the processing method, the infiltration time is 12 to 48 hours.

[0018] More preferably, in the preparation method, the infiltration time is 24 hours.

[0019] Furthermore, the drying method is: after soaking, leaving the liquid, taking the raw material and drying it once until the moisture content is below 20%; then mixing it with the liquid and drying it twice until the moisture content is below 5% (referred to as drying method a).

[0020] Furthermore, the drying method may also be: after soaking, drying the raw material until the moisture content is below 5% (referred to as drying method b).

[0021] Both drying methods described in the present invention can be used in actual production. The only difference is that in drying method a, after infiltration, the raw material is dried to a certain moisture content and then mixed with the infiltrating liquid and dried, which can further increase the infiltration time.

[0022] In the present invention, the sodium bicarbonate aqueous solution or sodium carbonate aqueous solution with a mass volume concentration of 0.5 to 10% is specifically prepared by adding 0.5 to 10 g of sodium bicarbonate or sodium carbonate to 100 mL of water.

[0023] Furthermore, the raw material and sodium bicarbonate aqueous solution or sodium carbonate aqueous solution are mixed according to a mass volume ratio of 1 g: (2 to 6) mL.

[0024] Preferably, the raw material and the sodium bicarbonate aqueous solution or the sodium carbonate aqueous solution are mixed at a mass volume ratio of 1 g: (2-5) mL; more preferably, the raw material and the sodium bicarbonate aqueous solution or the sodium carbonate aqueous solution are mixed at a mass volume ratio of 1 g: 5 mL.

[0025] Preferably, the soaking temperature is 65°C.

[0026] Furthermore, the drying is carried out at 100-140°C.

[0027] Preferably, the raw material of the tangerine peel or tangerine peel base is sliced ​​or cut into strips. As a preferred embodiment, in a specific embodiment of the present invention, slicing or cutting the raw material of the tangerine peel or tangerine peel base into strips before infiltration is beneficial to increase the contact area between the raw material and the liquid, so that the raw material of the tangerine peel or tangerine peel base can be better infiltrated.

[0028] Additionally, the present invention protects the application of the processing method in preparing synergistic and / or bitter-removing Huajuhong or Huajuhongtai processed products, as well as the processed products obtained.

[0029] The enhancement refers to the improvement of cough relieving, expectorant and lung protecting effects.

[0030] The present invention also protects the use of the processed product of Huajuhong or Huajuhongtai in the preparation of cough suppressant medicines.

[0031] In Example 12 of the present invention, the cough latency time and cough frequency of each group of mice were measured, and the antitussive activity of the processed Huajuhongtai tablets and Huajuhongtai tablets (unprocessed) was compared and evaluated. The experimental results showed that compared with Huajuhongtai tablets, the processed product prepared by the processing method of the present invention can enhance the antitussive activity of Huajuhong, which is specifically manifested in prolonging the cough latency period and suppressing the cough frequency.

[0032] The present invention also protects the use of the processed product of Huajuhong or Huajuhongtai in the preparation of expectorant medicine.

[0033] In Example 13 of the present invention, the expectorant activity of the processed Huajuhongtai tablets and Huajuhongtai tablets (unprocessed) was compared and evaluated by measuring the phenol red excretion in mice. The experimental results showed that compared with Huajuhongtai tablets, the processed product prepared by the processing method of the present invention can enhance the expectorant activity of Huajuhong.

[0034] The present invention also protects the use of the processed product of Huajuhong or Huajuhongtai in the preparation of lung protection medicines.

[0035] In Example 14 of the present invention, a comparative evaluation was conducted on the improvement effects of processed Huajuhongtai tablets and unprocessed Huajuhongtai tablets on lung injury induced by cigarette smoke extract E (CES). The experimental results showed that the processed Huajuhongtai tablets could alleviate lung injury, reduce the aggregation of inflammatory cells in the lungs, thereby alleviating inflammatory damage, and reduce the release of proinflammatory factors, thereby alleviating the inflammatory response in mice. It has a significant lung-protecting effect and is more effective than the unprocessed Huajuhongtai tablets.

[0036] The processed products prepared by the processing method of the present invention, and products prepared with other Chinese medicinal materials or raw materials with medicinal and edible properties, as well as various pharmaceutically acceptable excipients should also be within the scope of protection of the present invention.

[0037] The present invention has the following beneficial effects:

[0038] The present invention adopts sodium bicarbonate or sodium carbonate solution to soak and process tangerine peel or tangerine peel embryo at a certain temperature to obtain a processed product. The solution used is environmentally friendly and green, and meets the requirements for the safety of medicine and food. At the same time, the present invention experiments show that after processing using the processing method of the present invention, the content of its components changes significantly, and the efficacy of the prepared processed product is significantly improved, which is specifically reflected in: the efficacy of tangerine peel embryo in relieving cough, resolving phlegm and protecting lungs after processing is significantly improved compared with the unprocessed tangerine peel embryo; the processed tangerine peel embryo brewing liquid has a soft mouthfeel, no bitterness, and moderate sourness and sweetness. After component analysis, it is found that the dissolution amount of naringin, the main bitter substance of tangerine peel embryo, is significantly reduced after processing, and the dissolution amount of naringin dihydrochalcone, a sweet substance, is significantly increased. The synergistic and bitter-removing processing method of tangerine peel and tangerine peel embryo provided by the present invention can not only improve its efficacy and activity, but also effectively remove the bitterness in the raw materials, which is very beneficial for oral drinking of patients with cough, phlegm and lung injury. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 These are actual photographs of the original Huajuhong fetus product, the original Huajuhong fetus product, the processed Huajuhong fetus product, and the processed Huajuhong product of the present invention.

[0040] Figure 2 The present invention is a fluid extract of the original product CG of Huajuhongtai and the processed product PCG of Huajuhongtai; wherein Figure 2 A in the figure is a diagram of fluid extract. Figure 2 Figure B is an enlarged view of the fluid extract (at the same concentration, the original product CG on the left precipitates).

[0041] Figure 3 This is a comparison chart of the results of the cough-relieving experiment of the processed Huajuhong product of the present invention; Figure 3 A in the equation is cough latency time, Figure 3 The B in the equation is the number of coughs.

[0042] Figure 4 This is a comparison chart of the results of the phlegm-resolving experiment of the processed Huajuhong product of the present invention; Figure 4 A in the equation is phenol red excretion. Figure 4 The B in it is expectorant activity.

[0043] Figure 5 This is a comparison chart of the results of the processed tangerine peel product of the present invention on H&E sections of mouse lungs (magnification: 200×).

[0044] Figure 6 This is a comparison chart of the alveolar results of the processed tangerine peel product of the present invention in a cigarette smoke extract-induced experiment; Figure 6 A is the mean lining interval (MLI) of lung tissue. Figure 6 B is the mean alveolar number MAN; C is the inflammation score Inflammationscore.

[0045] Figure 7 The effect of the processed product of Huajuhong on the number of cells in the bronchoalveolar lavage fluid (BALF) of mice with lung injury; Figure 7 A is the total cell count in BALF, Figure 7 B in the figure is the number of BALF macrophages. Figure 7 C in the figure is the number of neutrophils in BALF.

[0046] Figure 8 This is a comparison chart of the results of the processed Huajuhong product of the present invention on inflammation in mice; Figure 8 A in the equation is the COX-2 content in BALF. Figure 8 B in the figure is the IFN-γ content in BALF, Figure 8 C is the TNF-α content in BALF, Figure 8 D in the equation is the COX-2 content in the lungs. Figure 8 E in the figure is the lung IFN-γ content, Figure 8 F in the expression is the content of TNF-α in lung.

[0047] Figure 9 This is a comparison chart of the effects of the processed Huajuhong product of the present invention on oxidative stress in mice; Figure 9 A in the equation is lung CAT content. Figure 9 B in the equation is lung GSH content. Figure 9 The C in it is the SOD content in the lungs.

[0048] Note: Figure 3 、 Figure 4 *P≤0.05 and **P≤0.01 indicate significant differences compared with the Model group; Figures 6 to 9 ##P≤0.01 indicates significant difference compared with the CON group, *P≤0.05 and **P≤0.01 indicate significant difference compared with the CSE group. DETAILED DESCRIPTION

[0049] The present invention will be further described below with reference to the accompanying drawings and specific examples, but the examples do not limit the present invention in any way. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art.

[0050] In the embodiments of the present invention, all used are Huajuhong or Huajuhong embryo obtained from authentic Huazhou pomelo of Huazhou origin.

[0051] Huajuhong decoction pieces: Huajuhong is cut into strips or blocks and dried in the sun.

[0052] Huajuhong fetal slices: Huajuhong fetal slices are round slices like deer antlers.

[0053] Unless otherwise specified, the reagents and materials used in the following examples were commercially available.

[0054] Example 1

[0055] Cut 200g of dried tangerine peel into strips and add 5 times its weight by volume of 0.5% sodium bicarbonate aqueous solution. Soak the dried tangerine peel at 65°C for 24 hours. Remove the dried tangerine peel and reserve the liquid. Dry the soaked dried tangerine peel at 120°C to a moisture content of 20%. Remove the dried tangerine peel and mix it with the liquid. Dry it at 120°C to a moisture content of 5% and set aside.

[0056] Example 2

[0057] Cut 200g of tangerine peel into slices, add 5 times its weight by volume of 2% sodium bicarbonate solution, and soak at 65°C for 24 hours. Remove the tangerine peel and save the liquid. Dry the soaked tangerine peel at 120°C to a moisture content of 20%, remove it, mix it with the liquid, and dry it at 120°C to a moisture content of 5%, and set aside.

[0058] Example 3

[0059] Add 5 times the weight volume of 5% sodium bicarbonate solution to 200g of tangerine peel slices and soak at 65°C for 24 hours. Remove the slices and save the liquid. Dry the soaked tangerine peel slices at 120°C to a moisture content of 20%. Remove and mix with the liquid again. Dry at 120°C to a moisture content of 5% and set aside.

[0060] Example 4

[0061] Add 5 times the weight volume of 2% sodium bicarbonate solution to 200g of Huajuhong slices. Soak at 90°C for 12 hours. Remove the Huajuhong slices and retain the liquid. Dry the soaked Huajuhong slices at 120°C to a moisture content of 20%. Remove, mix thoroughly with the liquid, and dry at 120°C to a moisture content of 5%. Set aside.

[0062] Example 5

[0063] Add 2 times the weight volume of 4% sodium bicarbonate aqueous solution to 200g of Huajuhong slices and soak at 50℃ for 48 hours. Take out Huajuhong slices and bake at 120℃ until the moisture content is 5% and set aside.

[0064] Example 6

[0065] Add 2 times the weight volume of 5% sodium carbonate aqueous solution to 200g of Huajuhong slices and soak at 90℃ for 24 hours. Take out Huajuhong slices and bake at 120℃ until the moisture content is 5% and set aside.

[0066] Example 7

[0067] Add 3 times the weight volume of 4% sodium bicarbonate aqueous solution to 200g of tangerine peel slices and soak at 65℃ for 24 hours. Take out the tangerine peel slices and bake at 120℃ until the moisture content is 5% and set aside.

[0068] Example 8

[0069] Add 3 times the weight volume of 4% sodium bicarbonate aqueous solution to 200g of tangerine peel sheet and soak at 90℃ for 24 hours. Take out the tangerine peel sheet and bake at 120℃ until the moisture content is 5% and set aside.

[0070] Example 9

[0071] Add 5 times the weight volume of 10% sodium bicarbonate aqueous solution to 200g of tangerine peel slices and soak at 90°C for 6 hours. Remove the slices and save the liquid. Dry the soaked tangerine peel slices at 120°C to a moisture content of 20%, remove them, mix them with the liquid, and dry them at 120°C to a moisture content of 5%. Set aside.

[0072] Example 10

[0073] Add 5 times the weight volume of 5% sodium bicarbonate solution to 200g of tangerine peel. Soak at 50°C for 72 hours. Remove the tangerine peel and save the liquid. Dry the soaked tangerine peel at 120°C to a moisture content of 20%. Remove and mix with the liquid. Dry at 120°C to a moisture content of 5% and set aside.

[0074] The color of the Huajuhong or Huajuhong embryo after processing of the present invention becomes obviously darker. Taking Examples 1 and 3 as examples, the color of Huajuhong embryo and Huajuhong after processing becomes obviously darker ( Figure 1 ).

[0075] Example 11 Analysis of changes in the composition of the processed product

[0076] 1. Experimental methods:

[0077] 1.1 Preparation of sample solution

[0078] The original product of Huajuhongtai tablets (GG) and the processed sample of Huajuhongtai tablets (PGG) prepared in the above example were weighed separately, and 70% methanol aqueous solution was added according to a 1:1 (V / V) ratio. The mixture was vortexed for 15 minutes and centrifuged at 12000 r / min and 4 ° C for 3 minutes. The supernatant was taken to obtain the test solution.

[0079] 1.2 Liquid quality conditions

[0080] Separation was performed using an Agilent SB-C18 column (1.8 μm, 2.1 mm × 100 mm). Phase A consisted of ultrapure water containing 0.1% formic acid, and phase B consisted of acetonitrile. The column oven temperature was 40°C, and the injection volume was 2 μL. Gradient elution was as follows: 0.0–9.0 min, 5% B to 95% B; 9.0–10.0 min, 95% B; 10.0–11.1 min, 95% B to 5% B; 11.1–14.0 min, 5% B.

[0081] 1.3 Ingredient identification

[0082] The components of the citrus aurantium were analyzed by extracting the molecular ion peaks of the main components in the mass spectrum and comparing them with the secondary mass spectra and LC / MS database of standards such as naringin, naringenin and naringin dihydrochalcone.

[0083] 2. Experimental results:

[0084] In this study, at the same dose, the fluidity of the PCG fluid extract of the Huajuhong tire tablets was better than that of the original product CG. The results are shown in the following example using the PCG fluid extract of the Huajuhong tire tablets of Example 3 as an example. Figure 2 shown.

[0085] Composition analysis revealed that the processed Huajuhongtai Tablet (PCG) is rich in flavonoids such as naringin, naringin, vitexin, apigenin, naringenin, and naringenin dihydrochalcone. Furthermore, a comparison of the compositional content of the processed Huajuhongtai Tablet (PCG) with that of the original CG revealed significant changes. Notably, after processing, the amount of naringin, one of the main active ingredients in Huajuhongtai Tablet (and also the primary bitter component in Huajuhongtai Tablet), was relatively reduced. The content of flavonoids such as naringenin-7-O-glucoside, naringenin, and naringenin dihydrochalcone increased. After the processing of Huajuhongtai Tablet, some naringin may become a more readily soluble active ingredient. This change in chemical composition may explain the change in the fluidity of the extract in this study. Furthermore, due to the significant reduction in the content of naringin, the main bitter component, the infusion of the processed Huajuhongtai Tablet (PCG) no longer has a bitter taste, but rather a moderate sweetness and sourness, resulting in a smooth and mellow taste. Taking the processed product PCG of Huajuhong tablets in Example 3 as an example, the results are shown in Table 1.

[0086] Table 1 Composition analysis of the original product CG and processed product PCG of Huajuhong

[0087]

[0088]

[0089] Example 12 Study on the Changes in Antitussive Efficacy of Processed Products

[0090] In this example, an antitussive efficacy evaluation experiment was conducted on the processed and unprocessed Huajuhongtai.

[0091] 1. Preparation method of animal drug solution: Take 200 g of Huajuhongtai tablets (GG) and 200 g of Huajuhongtai tablets prepared in Example 3 (PCG), add 8 times of water and boil twice, each time for 45 minutes, filter, combine the filtrate, and concentrate to 200 mL to obtain GG solution and PCG solution, respectively, and store them in the refrigerator for later use.

[0092] 2. Experimental animals and methods

[0093] After adaptive feeding, male ICR mice were randomly divided into 6 groups:

[0094] Model group: gavage with normal saline containing 0.5% CMC-Na;

[0095] Kebiqing group (Pen): Kebiqing (Pen) was administered orally at a dose of 0.25 g / kg;

[0096] Huajuhongtai tablets group (CG): administered GG solution by gavage, dose 1.0 g / kg;

[0097] Low-dose Huajuhongtai tablets preparation group (PCGL): PCG solution was administered orally at a dose of 0.5 g / kg;

[0098] Medium-dose processed tangerine peel tablets group (PCGM): PCG solution was administered orally at a dose of 1.0 g / kg;

[0099] High-dose Huajuhongtai tablets preparation group (PCGH): PCG solution was administered by gavage at a dose of 2.0 g / kg.

[0100] Each group received the drug once daily for seven consecutive days. One hour after the final dose, the mice were placed in a 500-mL glass chamber and exposed to 0.2 mL of ammonia solution. Observers recorded cough frequency and latency over a three-minute period.

[0101] 3. Experimental results:

[0102] This experiment compared and evaluated the antitussive (antitussive) activity of the processed products of Citrus aurantium by measuring the cough latency time and cough frequency of each group of mice. Indicates that the result is Figure 3 The experimental results showed that the processed product of Huajuhongtai tablets showed a more significant antitussive effect.

[0103] like Figure 3Middle A: Compared with the model group, Huajuhongtai Tablet CG and its processed product PCG significantly prolonged the cough latency. Moreover, at the same dose (1.0 g / kg), PCG significantly prolonged the cough latency compared with GG.

[0104] On the other hand, Figure 3 Figure B shows that ammonia stimulation caused mice to cough continuously (model group). Compared with the model group, both Huajuhongtai Tablet CG and its processed product PCG significantly suppressed the number of coughs. Specifically, at the same dose, Huajuhongtai Tablet PCG demonstrated superior antitussive activity compared to the original CG.

[0105] Therefore, it can be seen that the processed Huajuhongtai tablets prepared by the processing method of the present invention have an antitussive effect, and the effect is significantly better than that of the unprocessed Huajuhongtai tablets at the same dose.

[0106] Example 13 Study on the Changes in the Expectorant Efficacy of Processed Products

[0107] In this embodiment, an experiment was conducted to evaluate the expectorant efficacy of the processed Huajuhong and the unprocessed Huajuhong embryo.

[0108] 1. Preparation method of drug solution for animal administration: the same as that in Example 12.

[0109] 2. Experimental animals and methods

[0110] After adaptive feeding, male ICR mice were randomly divided into 6 groups:

[0111] Model group: gavage with normal saline containing 0.5% CMC-Na;

[0112] Ambroxol group (Amb): Ambroxol (Amb) was administered orally at a dose of 0.25 g / kg;

[0113] Huajuhongtai tablets group (CG): CG solution was administered orally at a dose of 1.0 g / kg;

[0114] Low-dose Huajuhongtai tablets preparation group (PCGL): PCG solution was administered orally at a dose of 0.5 g / kg;

[0115] Medium-dose processed tangerine peel tablets group (PCGM): PCG solution was administered orally at a dose of 1.0 g / kg;

[0116] High-dose Huajuhongtai tablets preparation group (PCGH): PCG solution was administered by gavage at a dose of 2.0 g / kg.

[0117] The above groups were administered once daily for seven consecutive days. After seven days, the mice were fasted for 24 hours but had free access to water for 24 hours. After 0.5 hours of normal gavage administration, each mouse was intraperitoneally injected with 0.5% phenol red in normal saline (10 mL / kg). After 0.5 hours, the mice were sacrificed, the tissue surrounding the trachea was peeled off, and a section of the trachea from the thyroid cartilage to the bronchial branch was cut. The trachea was placed in a 5 mL centrifuge tube, 1 mL of 5% NaHCO3 solution was added, vortexed, and then sonicated for 20 minutes. The supernatant was centrifuged at 1500 rpm for 10 minutes, the absorbance was measured at 546 nm, and the phenol red excretion was calculated according to the phenol red standard curve. The phenol red excretion of the model group was used as the control. The expectoration rate (%) = (corrected phenol red content of the treatment group / corrected phenol red content of the model group) × 100%.

[0118] 3. Experimental results:

[0119] This study compared and evaluated the expectorant activity of the Juhongtai tablets by measuring the phenol red excretion of mice in each group. Indicates that the result is Figure 4 The experimental results showed that phenol red secretion in the Huajuhongtai Pian processed PCG group was significantly increased and significantly superior to that in the Huajuhongtai Pian GG group. Oral administration of the Huajuhongtai Pian processed PCG group demonstrated a significant expectorant effect. At the same dose, the Huajuhongtai Pian processed PCG group significantly increased the expectorant rate, and its expectorant effect was even superior to that of the unprocessed Huajuhongtai Pian GG group. Therefore, it can be concluded that the Juhongtai Pian processed group exhibits an expectorant effect, demonstrating a stronger expectorant effect than the unprocessed Huajuhongtai Pian.

[0120] Example 14 Study on the Changes in Lung Protection Efficacy of Processed Products

[0121] In this example, the processed product PCG of Citrus aurantium and the unprocessed original product CG of Citrus aurantium were used to evaluate their efficacy against lung injury induced by cigarette smoke extract E (CSE).

[0122] 1. Preparation method of drug solution for animal administration: the same as that in Example 12.

[0123] 2. Experimental animals and methods

[0124] After adaptive feeding, male ICR mice were randomly divided into 7 groups:

[0125] The normal group (CON), model group (CSE), dexamethasone group (DEX, 0.5 mg / kg), unprocessed Huajuhongtai tablets group (CG, 1.0 g / kg), high-dose processed Huajuhongtai tablets group (PCGH, 2.0 g / kg), medium-dose processed Huajuhongtai tablets group (PCGM, 1.0 g / kg), and low-dose processed Huajuhongtai tablets group (PCGL, 0.5 g / kg) were fed according to the following method:

[0126] Normal group (CON): administered with normal saline via gavage;

[0127] Model group (CSE): mice were gavaged with normal saline and intraperitoneally injected with 0.2 mL of CSE on days 0, 5, 10, and 15.

[0128] Dexamethasone group (DEX): Dexamethasone (DEX) solution was administered orally at a dose of 0.5 mg / kg, and 0.2 mL of CSE was injected into the peritoneum of the mice on days 0, 5, 10, and 15, respectively;

[0129] Huajuhongtai tablets group (CG): GG solution was administered orally at a dose of 1.0 g / kg; and 0.2 mL of CSE was injected into the peritoneum of the mice on days 0, 5, 10, and 15, respectively.

[0130] High-dose Huajuhongtai tablets (PCGH) group: PCG solution was administered orally at a dose of 2.0 g / kg, and 0.2 mL of CSE was injected into the peritoneum of the mice on days 0, 5, 10, and 15, respectively.

[0131] Medium-dose processed tangerine peel tablets (PCGM) group: PCG solution was administered orally at a dose of 1.0 g / kg, and 0.2 mL of CSE was injected into the peritoneum of the mice on days 0, 5, 10, and 15, respectively.

[0132] Low-dose Huajuhongtai tablets preparation group (PCGL): PCG solution was administered orally at a dose of 0.5 g / kg, and 0.2 mL CSE was injected into the peritoneum of the mice on days 0, 5, 10, and 15, respectively.

[0133] Each group received the drug once daily for 20 consecutive days. During the experiment, the animals' diet was monitored daily. On the final day of the experiment, bronchoalveolar lavage fluid (BALF) and lung tissue were collected for subsequent pathological observation and inflammatory factor detection.

[0134] Lung tissue was obtained for HE staining and pathological observation and scoring. Pathological scoring criteria: a cross was drawn in the center of each visual field, and the number of alveolar septa (A) and the number of alveoli in each visual field (B) that intersected with the line were calculated. The total length of the cross line (L) and the area of ​​each visual field (S) were also measured. The mean lining interval (MLI, reflecting the mean diameter of the alveoli) and the mean number of alveoli (MAN, reflecting the alveolar density) of the lung tissue were calculated according to the following formulas: MLI = L / A and MAN = B / S; 4 different visual fields were selected for each section and scored using the corresponding inflammatory infiltration scoring criteria (scoring criteria: 0 points, no inflammatory infiltration; 1 point, inflammatory infiltration less than 25%; 2 points, inflammatory infiltration 25-50%; 3 points, inflammatory infiltration 50-75%; 4 points, inflammatory infiltration more than 75%). Statistical data are presented as mean ± standard deviation. P < 0.05 indicates that the difference between the groups is statistically significant.

[0135] 2. Experimental results:

[0136] H&E staining Figure 5 As shown in the figure, the lung tissues of mice in the CSE group showed obvious emphysema and inflammatory changes, which were manifested as alveolar hemorrhage, severe damage to alveolar structure, alveolar expansion and fusion, widening of alveolar septa, and infiltration of inflammatory cells. In contrast, the PCG group of the Huajuhongtai tablets preserved the alveolar structure, and reduced pulmonary interstitial congestion, edema and inflammation, just like the DEX group, while the original CG group still had a small amount of infiltrating inflammatory cells. Figure 6 As shown in the results, compared with the CON group, the mean lining interval (MLI) and inflammation score (Inflammation score) of the lung tissue of mice in the CSE group were significantly increased, while the mean alveolar number (MAN) was significantly decreased, further indicating that CSE induction can lead to severe alveolar damage and lung inflammatory infiltration (P<0.01). When PCG was used to treat mice with lung injury, these pathological lesions in the lung tissue were effectively alleviated. At the same dose, the effect of the processed Huajuhongtai tablet PCG on reducing MLI and inflammation score and increasing the mean alveolar number (MAN) was significantly better than that of the unprocessed Huajuhongtai tablet GG. At the same time, the high-dose effect of the processed Huajuhongtai tablet PCG was better than that of the DEX-positive drug group.

[0137] Combine Figure 7 The changes in BALF cell count and inflammatory factors in the CSE group indicated that the lungs of mice were damaged. Figure 7As shown in Figure A, the total cell number in the alveolar lavage fluid (BALF) of the CSE group was significantly higher than that of the normal CON group (P<0.01). After administration of CG, PCG and the positive drug DEX, the total cell number in the BALF of mice showed a decreasing trend. Among them, after intervention with different concentrations of PCG, the total cell number was significantly decreased compared with the CSE group (P<0.01); although the total cell number decreased after the positive drug DEX and the unprocessed Huajuhongtai tablets (CG), there was no significant difference with the CSE group (P>0.05).

[0138] In addition, this study further identified and analyzed different cells in the BALF. Figure 7 As shown in Figures B and C, the number of macrophages and neutrophils in the CSE group was significantly increased compared with the CON group (P < 0.01). However, after oral administration of PCG, the number of macrophages in the mouse bronchoalveolar lavage fluid (BALF) was significantly decreased (P < 0.01). These results indicate that macrophages predominate in the mouse lungs and that PCG, a processed product of Huajuhongtai tablets, can reduce the accumulation of inflammatory cells in the lungs, thereby alleviating inflammatory damage.

[0139] During lung inflammation infiltration, macrophages have the function of amplifying the inflammatory response by releasing pro-inflammatory cytokines (such as COX-2, IFN-γ and TNF-α). Figure 8 As shown in the results, compared with the CON group, the levels of COX-2, IFN-γ and TNF-α in the BALF and lungs of the CSE group were significantly increased (all P<0.01), indicating that the mouse model was successfully established. However, compared with the CSE group, treatment with DEX, CG and PCG significantly reversed these changes. The levels of proinflammatory factors (COX-2, IFN-γ and TNF-α) in the three PCG dose groups were significantly reduced. Moreover, the improvement effects of the dose group and the high-dose group were better than those of the positive drug group DEX and the unprocessed Huajuhongtai tablets (CG). The results showed that the processed Huajuhongtai tablets PCG can reduce the release of proinflammatory factors to alleviate the pulmonary inflammatory response in mice, and the effect is better than the unprocessed Huajuhongtai tablets CG at the same dose.

[0140] like Figure 9As shown in the results, compared with the CON group, CSE-induced lung oxidative stress was accompanied by significant decreases in the activities of the antioxidant enzymes CAT, GSH, and SOD (P < 0.01). However, compared with the CSE group, administration of DEX, CG, and PCG increased the levels of CAT, GSH, and SOD to a certain extent (P < 0.01, P < 0.05, and P < 0.01, respectively). Furthermore, at the same dose, PCG was more effective than CG in increasing the levels of CAT, GSH, and SOD. These results suggest that PCG, a processed product of Huajuhongtai Tablet, can alleviate oxidative stress in the lungs and exert a lung-protective effect, with superior efficacy compared to unprocessed Huajuhongtai Tablet CG at the same dose.

[0141] The above results show that the processed Huajuhongtai tablets (PCG) have a significant lung protection effect, and the effect is better than the unprocessed Huajuhongtai tablets (CG).

[0142] The above embodiments are preferred implementations of the present invention, but the implementations of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be considered equivalent replacement methods and are included in the scope of protection of the present invention.

Claims

1. A method for preparing Huajuhong and Huajuhong embryo, which is used in preparing Huajuhong or Huajuhong embryo processed products that enhance efficacy and / or eliminate bitterness; The steps of the preparation method are as follows: Take the raw material of Huajuhong or Huajuhong embryo, mix it with a sodium bicarbonate aqueous solution or a sodium carbonate aqueous solution with a mass volume concentration of 0.5-10%, soak it at 50-90°C for 6-72 hours, and then dry it to obtain a processed product; in, The weight-to-volume ratio of the raw material to the sodium bicarbonate aqueous solution or the sodium carbonate aqueous solution is 1 g: (2-6) mL; The enhancement refers to the enhancement of cough relieving, expectorant and lung protecting effects; The drying method is as follows: after soaking, the liquid is left, the raw material is dried once until the moisture content is below 20%; then the raw material is mixed with the liquid and dried twice until the moisture content is below 5%.

2. The application according to claim 1, characterized in that In the preparation method, the mass volume concentration of the sodium bicarbonate aqueous solution or the sodium carbonate aqueous solution is 0.5-5%.

3. The application according to claim 1, characterized in that In the processing method, the infiltration time is 12 to 48 hours.

4. The application according to claim 3, characterized in that In the processing method, the infiltration time is 24 hours.

5. The application according to claim 1, characterized in that: In the preparation method, the weight-to-volume ratio of the raw material to the sodium bicarbonate aqueous solution or the sodium carbonate aqueous solution is 1 g: (2-5) mL.

6. The application according to claim 1, characterized in that: In the preparation method, the weight-to-volume ratio of the raw material to the sodium bicarbonate aqueous solution or the sodium carbonate aqueous solution is 1 g:5 mL.

7. The use according to claim 1, characterized in that In the processing method, the infiltration temperature is 65°C.

8. The application according to claim 1, characterized in that: In the processing method, the drying is performed at 100-140°C.

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