Composition for treating yin deficiency and phlegm stasis syndrome as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to a composition for treating yin deficiency and phlegm stasis syndromes as well as a preparation method and application thereof. The composition is prepared from the following medicinal raw materials: rhizoma bletillae, thunberg fritillary bulb, platycodon grandiflorum, radix stemonae, lily, bryozoatum, radix asteris, pseudo-ginseng, Chinese mahonia stem and liquorice. The preparation method comprises the following steps: (1) adding alcohol into rhizoma bletillae, pseudo-ginseng, Chinese mahonia stem and liquorice, and performing reflux extraction to obtain an alcohol extract and decoction dregs; (2) mixing platycodon grandiflorum, thunberg fritillary bulb, radix stemonae, lily, bryozoatum and radix asteris, adding water, decocting, performing microwave treatment, and filtering to obtain an aqueous extract; (3) mixing the medicine residue and the water extract, performing enzymolysis, decocting, and centrifuging to obtain supernate; and (4) mixing the alcohol extract and the supernate, concentrating and drying to obtain the composition. A specific preparation method is adopted, the effects of active ingredients in the traditional Chinese medicine components can be fully exerted, and the treatment score evaluation on the yin deficiency and phlegm stasis syndrome is remarkably improved.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of traditional Chinese medicine, and particularly relates to a composition for treating yin deficiency and phlegm stasis, a preparation method and application thereof. BACKGROUND

[0002] Chronic obstructive pulmonary disease (COPD) is a chronic airway inflammatory disease characterized by persistent airflow limitation, which is not completely reversible and has a significant adverse impact on the daily life of patients. The main clinical manifestations of COPD are cough, sputum, and dyspnea, and acute exacerbation often occurs during the course of the disease, which is the main factor promoting the continuous progression of the disease. COPD belongs to the category of "lung distension" in traditional Chinese medicine. The main symptoms of "lung distension" described by physicians of past dynasties are mainly summarized as chest fullness, stuffy as if blocked, wheezing and coughing, excessive sputum, and restlessness, palpitation, etc., that is, the six characters of "cough, sputum, wheezing, distension, swelling, and stasis" are the main characteristics. In the acute exacerbation period of chronic obstructive pulmonary disease (COPD), although the effect of traditional Chinese medicine treatment is not as rapid as that of western medicine, in the stable period, the treatment with traditional Chinese medicine has obvious advantages in terms of improving symptoms, reducing acute exacerbation, improving exercise capacity and quality of life, etc.

[0003] The stable period of COPD includes two main types of lung and kidney deficiency and phlegm stasis blocking the lung. At present, Chinese invention patent application CN119345319A discloses a traditional Chinese medicine compound for treating chronic obstructive pulmonary disease in the stable period and its application, wherein the traditional Chinese medicine composition contains Herba Epimedii, Herba Cuspariae, Radix Paeoniae Alba, Radix Salviae Miltiorrhizae, Lumbricus, Rhizoma Dispori, Radix Ophiopogonis, Radix Scrophulariae, Radix Paeoniae Rubra, Rhizoma Zingiberis Recens, Cortex Mori, Rhizoma Pinelliae, Rhizoma Coptidis, and Semen Ginkgo. The prescription mainly warms yang and tonifies qi (Herba Epimedii, Herba Cuspariae, Radix Paeoniae Alba), and combines with clearing heat and reducing phlegm (Radix Paeoniae Alba, Cortex Mori) and promoting blood circulation to remove meridian obstruction (Radix Salviae Miltiorrhizae, Lumbricus, Rhizoma Dispori). However, there is a risk of warming and drying yin: Herba Epimedii, Herba Cuspariae, and Rhizoma Zingiberis Recens, which are yang-warming drugs, account for a large proportion, and long-term use may consume lung yin, especially for patients with yin deficiency type COPD (such as red tongue with little moss, dry cough without sputum), which may aggravate the symptoms; although Radix Ophiopogonis and Radix Scrophulariae can nourish yin, the dosage is relatively weak compared with yang-warming drugs, and it is difficult to completely balance them. In addition, Rhizoma Pinelliae, Rhizoma Zingiberis Recens, and Rhizoma Coptidis may stimulate the gastrointestinal tract, and easily cause abdominal distension in patients with weak spleen and stomach. The dosage of each medicinal material in the prescription has a large span, and the compatibility is questionable.

[0004] Chinese invention patent application CN109939172A discloses a traditional Chinese medicine composition for chronic obstructive pulmonary disease in the stable period, which has the effects of tonifying lung, spleen and kidney qi, reducing phlegm and dampness, promoting blood circulation to remove blood stasis, and nourishing yin and clearing heat, and can improve the pathology of stable chronic obstructive pulmonary disease, but the improvement of specific traditional Chinese medicine syndrome manifestations is not clear.

[0005] Chinese invention patent application CN119139398A discloses a traditional Chinese medicine compound for treating stable chronic obstructive pulmonary disease with lung-kidney deficiency syndrome, a preparation method and application thereof. The raw materials of the traditional Chinese medicine compound include radix codonopsis, radix morindae officinalis, radix paeoniae alba, radix atractylodis macrocephalae, radix rehmanniae preparata, semen litchi, ramulus cinnamomi, radix salviae miltiorrhizae and radix ephedrae praeparata. The medicine formula focuses on tonifying lung and kidney (radix codonopsis, radix morindae officinalis and radix rehmanniae preparata), supplementing qi and activating blood (radix paeoniae alba and radix salviae miltiorrhizae), and is assisted by pungent and warm herbs for dispersing lung (radix ephedrae praeparata and ramulus cinnamomi). Among them, the side effects of pungent and warm herbs are prominent, radix ephedrae praeparata contains ephedrine, which may induce palpitation and elevated blood pressure, and the risk is higher for COPD patients with cardiovascular diseases; ramulus cinnamomi is pungent and warm, which may aggravate hot flashes and night sweats in people with internal heat due to yin deficiency. In the medicine formula, only one herb, radix rehmanniae preparata, is used to nourish yin and moisten lung, and the dosage is low, so the effect on COPD with lung-kidney yin deficiency (such as wheezing with dry throat and little sticky sputum) is limited.

[0006] Therefore, it is necessary to develop a traditional Chinese medicine composition for treating yin deficiency and phlegm stasis syndrome (stable chronic obstructive pulmonary disease) with improved drug effectiveness and safety and remarkable curative effect. SUMMARY

[0007] In view of the deficiencies of the prior art, the present application provides a composition for treating yin deficiency and phlegm stasis syndrome and a preparation method and application thereof, which can effectively solve the problem of medicine for treating chronic obstructive pulmonary disease.

[0008] To achieve the above-mentioned object, the technical scheme adopted by the present application is as follows: A preparation method of a composition for treating yin deficiency and phlegm stasis syndrome, the composition comprising the following raw medicinal materials: radix bletillae, fritillaria thunbergii miq., platycodon grandiflorum, stemona tuberosa, lilium brownii, sea stone, radix pini, radix notoginseng, and radix peganum harmala and licorice; The preparation method comprises the following steps: (1) reflux extraction of radix bletillae, radix notoginseng, radix peganum harmala and licorice with alcohol to obtain alcohol extract and dregs; (2) mixing fritillaria thunbergii miq., platycodon grandiflorum, stemona tuberosa, lilium brownii, sea stone and radix pini, and then performing microwave treatment after decocting with water to obtain water extract; (3) mixing the dregs and the water extract, and then performing enzymolysis and decoction, and then centrifuging to obtain supernatant; (4) mixing the alcohol extract and the supernatant, and then concentrating and drying to obtain the composition.

[0009] Preferably, in step (1), the alcohol is ethanol with a volume concentration of 50%-70%; the reflux extraction is performed at a temperature of 45-60°C for 1-3h, and the solid-liquid ratio is 1:20-40g / mL.

[0010] Preferably, in step (2), the solid-liquid ratio of decoction is 1:10-25g / mL, and the decoction time is 1-3h; the microwave treatment is performed at a power of 500-800W for 5-15min.

[0011] Preferably, the enzymes used in the enzymolysis in step (3) are cellulase, beta-xylosidase and yeast extract enzyme, and the mass ratio of the cellulase, beta-xylosidase and yeast extract enzyme is 1:0.5-1:0.1-0.3.

[0012] Further preferably, the mass ratio of the cellulase, beta-xylosidase and yeast extract enzyme is 1:0.8:0.2.

[0013] Preferably, the amount of the enzyme added in step (3) is 0.3%-1% of the mass of the water extract, the temperature of the enzymolysis is 50-55℃, the time of the enzymolysis is 2-6h, and the pH of the enzymolysis is 5.5-6.5.

[0014] Preferably, the time of the decoction in step (3) is 0.5-1.0h.

[0015] The present application also relates to the composition for treating yin deficiency and phlegm stasis syndrome prepared by the preparation method, and the raw materials of the composition are 110-120 parts of white, 35-45 parts of Zhejiang fritillary, 65-70 parts of platycodon, 65-70 parts of stemona, 65-70 parts of lily, 65-70 parts of sea sponge, 65-70 parts of smilax, 35-45 parts of sanqi, 65-70 parts of mahonia, and 15-25 parts of licorice, in terms of weight.

[0016] The present application also relates to the application of the composition for treating yin deficiency and phlegm stasis syndrome prepared by the preparation method in the preparation of traditional Chinese medicine preparations for treating yin deficiency and phlegm stasis syndrome.

[0017] Preferably, the dosage form of the traditional Chinese medicine preparation is selected from any one of granules, tablets, capsules, pills, powders, syrup, oral solution, oral suspension, oral emulsion, mixture and decoction.

[0018] White, the dried tuber of the liliaceae plant white Bletilla striata (Thunb.) Reichb. f. has the effect of moistening the lungs, reducing the viscosity of respiratory mucus to relieve cough and reduce phlegm, clearing dry heat, soothing the liver and regulating qi.

[0019] Zhejiang fritillary, the bulb of the liliaceae plant Zhejiang fritillary, has the effects of clearing heat and moistening the lungs, detoxifying and reducing swelling, relieving cough and asthma.

[0020] Platycodon, the root of the platycodon grandiflorum, is used to treat external cough, sore throat, lung abscess, chest fullness, and abdominal pain.

[0021] Stemona, the bulb of the stemonaceae plant straight stemona Stemona sessilifolia (Miq.) Miq., stemona tuberosa Stemona japonica (Bl.) Miq. or stemona tuberosa Stemona tuberosaThe dried tuberous root of Lour. It has anti-tuberculosis and antitussive / asthmatic effects.

[0022] Lily refers to the dried, fleshy scales of the lily family plants *Lilium tigrinum*, *Lilium tigrinum*, or *Lilium davidii*. It is primarily used to treat chronic cough due to lung dryness or yin deficiency, coughing up blood; lingering fever after febrile diseases, restlessness, palpitations, and mental confusion; and beriberi with edema.

[0023] The pumice mentioned is the skeleton of bryozoans such as the Bryozoa bream and the Bryozoa bursa-palatineids; it has the effects of clearing lung heat, resolving phlegm, softening hard masses, and promoting urination. According to "Ben Cao Zheng", pumice can aid digestion, clear heat phlegm, relieve fever, thirst, and urinary tract infections, stop cough and wheezing, and promote diuresis.

[0024] Aster tataricus is the root and rhizome of the plant Asteraceae family. It is primarily used to treat coughs and asthma due to wind-cold, coughs with purulent sputum and blood due to deficiency, sore throat, and difficulty urinating.

[0025] Panax notoginseng is the tuberous root of Panax notoginseng, a plant belonging to the genus Panax notoginseng in the family Araliaceae. It is mainly used to treat hematemesis, hemoptysis, epistaxis, hematochezia, bloody dysentery, metrorrhagia, abdominal masses, postpartum hemorrhage, lochia retention, traumatic bleeding, external bleeding, and carbuncles and boils with pain.

[0026] Mahonia bealei (Fort.) Carr. or Mahonia fortunei (Lindl.) Fedde, both belonging to the Berberidaceae family, are dried stems. They enter the liver, stomach, and large intestine meridians. They clear heat and dry dampness, purge fire and detoxify. They are used for damp-heat diarrhea, jaundice, red and swollen eyes, toothache due to stomach heat, boils, carbuncles, dysentery; and jaundice-type hepatitis.

[0027] Licorice, is the plant of the genus Glycyrrhiza in the legume family. Glycyrrhiza uralensis The roots and rhizomes of Fisch. Enter the spleen, stomach, and lung meridians. Primarily used to treat spleen and stomach weakness, poor appetite, abdominal pain and loose stools, fatigue and fever, pulmonary tuberculosis and cough, palpitations, and epilepsy; when used raw, it treats sore throat, peptic ulcers, carbuncles and sores, and detoxifies drug poisoning and food poisoning.

[0028] Compared with the prior art, the present invention has the following beneficial effects: This invention employs a specific preparation method, in which some medicinal materials are extracted with alcohol to obtain an ethanol extract and residue. The residue is then mixed with the water extracts of Platycodon grandiflorus, Fritillaria thunbergii, Stemona japonica, Lilium brownii, pumice, and Aster tataricus to prepare a traditional Chinese medicine composition. This composition can fully exert the effects of the active ingredients in each medicinal component and significantly improve the treatment score evaluation for Yin deficiency and phlegm stasis syndrome. Detailed Implementation

[0029] The following non-limiting examples can enable those of ordinary skill in the art to more fully understand the present application, but are not intended to limit the present application in any way. The following description is merely exemplary of the application and is not intended to limit the scope, applicability or configuration of the application in any way. Various changes and modifications can be made to the application by those of ordinary skill in the art without departing from the scope of the application as described by the claims.

[0030] The present application is further described in the following specific examples. The various instruments, devices, equipment, reagents, products, etc. used in the examples of the present application are obtained from commercial sources unless otherwise stated.

[0031] Cellulase (10,000 u / g), β-xylosidase (1-20,000 u / g), and yeast extract enzyme (comprehensive enzyme activity), all purchased from Nanning Dongheng Huadao Biological Technology Co., Ltd.

[0032] Example 1 A composition for treating yin deficiency and phlegm stasis, the medicinal material raw materials of the composition comprise, by weight fraction: 118 parts of white bletilla, 40 parts of Zhebei, 67 parts of jujube, 67 parts of trichosanthes, 67 parts of lily, 67 parts of sea stone, 67 parts of purple root, 40 parts of sanqi, 67 parts of mahonia, and 20 parts of licorice.

[0033] The preparation method is as follows: (1) Mix white bletilla, sanqi, mahonia, and licorice, and reflux extract with 60% volume concentration ethanol, specifically reflux extract at 55℃ for 2h, with a solid-liquid ratio of 1:30g / mL, to obtain an alcohol extract and dregs.

[0034] (2) Mix jujube, Zhebei, trichosanthes, lily, sea stone, and purple root, and decoct with water at a solid-liquid ratio of 1:20g / mL for 2h, then microwave treat for 10min at a microwave power of 600W, and filter to obtain a water extract.

[0035] (3) Mix the dregs and the water extract, and enzymatically hydrolyze at 50℃ and pH 6.0 for 4h, with an enzyme addition amount of 0.5% of the mass of the water extract, then decoct for 0.5h, and centrifuge to obtain a supernatant; The enzymes used for enzymatic hydrolysis are cellulase, β-xylosidase, and yeast extract enzyme at a mass ratio of 1:0.8:0.2.

[0036] (4) Mix the alcohol extract and the supernatant, concentrate, and dry to obtain the composition.

[0037] Example 2 A composition for treating yin deficiency and phlegm stasis, the medicinal material raw material composition of the composition comprises, by weight fraction: 118 parts of white and 40 parts of Zhebei, 67 parts of Jiegeng, 67 parts of Baibu, 67 parts of Baihe, 67 parts of Haifushi, 67 parts of Ziyuan, 40 parts of Sanqi, 67 parts of Gonglaomu, and 20 parts of Gancao.

[0038] The preparation method is as follows: (1) Mix white and Sanqi, Gonglaomu and Gancao, add 50% by volume concentration ethanol for reflux extraction, specifically 60℃ reflux extraction for 1h, solid-liquid ratio is 1:20g / mL, to obtain alcohol extract and dregs.

[0039] (2) Mix Jiegeng, Zhebei, Baibu, Baihe, Haifushi and Ziyuan, add water for decocting 1h according to solid-liquid ratio 1:10g / mL, then microwave treatment for 15min, microwave power is 500W, filter to obtain water extract.

[0040] (3) Mix dregs and water extract, enzymolysis for 2h at 55℃ and pH 5.5, enzyme addition amount is 0.8% of water extract mass, then decoct for 0.8h, centrifugal to obtain supernatant; The enzyme used in enzymolysis is cellulase, β-xylosidase and yeast extractase with mass ratio of 1:0.5:0.1.

[0041] (4) Mix alcohol extract and supernatant, concentrate, dry, to obtain the composition.

[0042] Example 3 A composition for treating yin deficiency and phlegm stasis, the medicinal material raw material composition of the composition comprises, by weight fraction: 118 parts of white and 40 parts of Zhebei, 67 parts of Jiegeng, 67 parts of Baibu, 67 parts of Baihe, 67 parts of Haifushi, 67 parts of Ziyuan, 40 parts of Sanqi, 67 parts of Gonglaomu, and 20 parts of Gancao; The preparation method is as follows: (1) Mix white and Sanqi, Gonglaomu and Gancao, add 70% by volume concentration ethanol for reflux extraction, specifically 45℃ reflux extraction for 3h, solid-liquid ratio is 1:40g / mL, to obtain alcohol extract and dregs.

[0043] (2) Mix Jiegeng, Zhebei, Baibu, Baihe, Haifushi and Ziyuan, add water for decocting 3h according to solid-liquid ratio 1:25g / mL, then microwave treatment for 5min, microwave power is 800W, filter to obtain water extract.

[0044] (3) Mix dregs and water extract, enzymolysis for 6h at 50℃ and pH 6.5, enzyme addition amount is 0.3% of water extract mass, then decoct for 1.0h, centrifugal to obtain supernatant; The enzyme used in the enzymolysis is cellulase, β-xylosidase and yeast extract enzyme with a mass ratio of 1:1:0.3.

[0045] (4) The alcohol extract and the supernatant are mixed, concentrated and dried to obtain the composition.

[0046] Comparative Example 1 The difference between this comparative example and Example 1 is only in the preparation method, and the specific preparation method is as follows: (1) All medicinal material raw materials are mixed, and water is added at a solid-liquid ratio of 1:20 g / mL for decoction for 2 h, and then microwave treatment is performed for 10 min at a microwave power of 600 W, and filtration is performed to obtain a water extract.

[0047] (2) The water extract is subjected to enzymolysis, specifically: enzymolysis is performed at 50°C and pH 6.0 for 4 h, and the enzyme is added in an amount of 0.5% of the mass of the water extract; then decoction is performed for 0.5 h, and centrifugation is performed to obtain a supernatant; The enzyme used in the enzymolysis is cellulase, β-xylosidase and yeast extract enzyme with a mass ratio of 1:0.8:0.2.

[0048] (3) The alcohol extract and the supernatant are mixed, concentrated and dried to obtain the composition.

[0049] Comparative Example 2 The difference between this comparative example and Example 1 is only in the preparation method, and the specific preparation method is as follows: (1) All medicinal material raw materials are mixed, and ethanol with a volume concentration of 60% is used for reflux extraction, specifically: reflux extraction is performed at 55°C for 2 h at a solid-liquid ratio of 1:30 g / mL to obtain an alcohol extract and a residue.

[0050] (2) Water is added to the residue at a solid-liquid ratio of 1:20 g / mL, and enzymolysis is performed at 50°C and pH 6.0 for 4 h, and the enzyme is added in an amount of 0.5% of the mass of the water, and then decoction is performed for 0.5 h, and centrifugation is performed to obtain a supernatant; The enzyme used in the enzymolysis is cellulase, β-xylosidase and yeast extract enzyme with a mass ratio of 1:0.8:0.2.

[0051] (3) The alcohol extract and the supernatant are mixed, concentrated and dried to obtain the composition.

[0052] Comparative Example 3 The difference between this comparative example and Example 1 is only in the enzyme in step (3), specifically: only cellulase is used in this comparative example.

[0053] Comparative Example 4 The difference between this comparative example and Example 1 is only in the enzyme in step (3), specifically: the enzyme used in this comparative example is β-xylosidase and yeast extract enzyme with a mass ratio of 1.6:0.4.

[0054] Comparative Example 5 The only difference between this comparative example and Example 1 is the enzyme in step (3), specifically: the enzyme used in this comparative example is cellulase and β-xylosidase in a mass ratio of 1:1.

[0055] Comparative Example 6 The only difference between this comparative example and Example 1 is the enzyme in step (3), specifically: the enzyme used in this comparative example is cellulase and yeast extractase in a mass ratio of 1:1.

[0056] Comparative Example 7 The only difference between this comparative example and Example 1 is the preparation method, specifically: All medicinal material raw materials were crushed, mixed, and added with water in a solid-liquid ratio of 1:20 g / mL, and then subjected to enzymatic hydrolysis at 50°C and pH 6.0 for 4 h, with the enzyme being added in an amount of 0.5% of the mass of the medicinal materials, and the enzyme used in the enzymatic hydrolysis being cellulase, β-xylosidase, and yeast extractase in a mass ratio of 1:0.8:0.2; then subjected to decoction for 0.5 h, centrifuged, and the supernatant was obtained, concentrated, and dried to obtain the composition.

[0057] Effect Test Effect Test 1 Treatment of Chronic Obstructive Pulmonary Disease 1.1 Animals: SPF level SD rats 130, half male and half female, body weight 180-200 g, feeding environment: room temperature 22-26°C, relative humidity 45%-65%, standard feed, free access to food and water.

[0058] 1.2 Reagents and Instruments: Hongqi Canal brand filter tip cigarette (flue-cured tobacco type, containing 11 mg of tar, 0.9 mg of smoke nicotine, and 11 mg of smoke carbon monoxide, Henan Anyang Cigarette Factory). IL-1β (EK0393), TNF-α (EK0526), MMP-9 (EK1463), and TIMP-1 (EK0583) enzyme-linked immunosorbent assay (ELISA) kits were purchased from Wuhan Dr. Deki Biological Engineering Co., Ltd.

[0059] 1.3 Model Preparation: SPF level SD rats were adaptively fed for 7 days. Randomly divided into blank group, model group, examples 1-3 group, comparative examples 1-7 group and Baibei Yifei capsule group, 10 in each group. The COPD lung qi deficiency syndrome rat model was copied by smoke combined with LPS (lipopolysaccharide) tracheal instillation and forced swimming method. The rats were placed in a constant temperature water tank (43±1) °C before smoking every day, forced to swim for 30 minutes, to consume the lung qi, then placed in a self-made smoking box, lit a cigarette, the smoke concentration reached 3000±500ppm, smoked every day, 1 time per day, 30 minutes each time, a total of 28 days of smoking, on the 1st day and the 14th day, the trachea was exposed by operation, 200 μL of LPS (1 mg / mL) was instilled into the trachea of each rat, and the trachea was not smoked on the same day, on the 22nd day, 3 times a day.

[0060] 1.4 Administration method: After the model was successfully established, the mice in examples 1-3 group, comparative examples 1-7 group and Baibei Yifei capsule group were orally administered with the corresponding dose of composition or drug, the administration amount was 0.4 g / kg, and the administration volume was 10 mL / kg. After 28 days of modeling, the administration was started, and the composition prepared in examples 1-3, comparative examples 1-7 and Baibei Yifei capsule were configured with distilled water to the concentration required by the administration volume every day after modeling. The blank group and the model group were administered with distilled water with the same volume, 1 time / day, and the administration was continuously given for 8 weeks. The body weight was weighed every week to adjust the drug amount.

[0061] 1.5 Index detection: The mental state, activity, fur, secretion, food and water intake changes, stool and urine, and symptoms such as cough, asthma, body weight of the rats were observed.

[0062] BALF IL-1β, TNF-α, MUC5AC level determination: ELISA method was used to detect the expression of IL-1β, TNF-α and MUC5AC in BALF, and the kit instructions were followed.

[0063] MMP-9, TIMP-1 expression in serum: ELISA method was used to detect the expression changes of MMP-9 and TIMP-1 in the serum of rats in each group, and the kit instructions were followed.

[0064] 1.6 Statistical analysis: The experimental data were statistically analyzed by using statistical software SPSS 25.0. The normality test was performed on the measurement data, the indicators conforming to the normal distribution were represented by, the one way ANOVA test was used for comparison among multiple groups, the LSD-t test was used for comparison between groups, and P<0.05 indicated that the difference was statistically significant.

[0065] 1.7 Experimental results: (1) General condition of rats The control group rats had shiny fur, good spirits, and normal food and water intake. The model group rats had dull fur, emaciation, lethargy, reduced activity, reduced food and water intake, loose stools, and wet bedding. The rats had reduced resistance when grabbed, and different degrees of panting, nasal mucus secretion, and other symptoms. After drug intervention, the rats' hair gradually became denser, shiny, and their food intake and body weight increased, and their overall activity improved.

[0066] (2) Expression of IL-1β, TNF-α, and MUC5AC in BALF Compared with the blank group, the expression of IL-1β, TNF-α, and MUC5AC in the BALF of the model group rats increased (P<0.05 or P<0.01); compared with the model group, the expression of IL-1β, TNF-α, and MUC5AC in the example and comparative example groups decreased (P<0.05 or P<0.01), and there was a significant difference between the comparative example and Example 1 groups (P<0.05). See Table 1 for details.

[0067]

[0068] Note: Compared with the blank group, & P<0.05, && P<0.01; compared with the model group, # P<0.05, ## P<0.01; compared with the Example 1 group, ▲ P<0.05.

[0069] (2) Expression of MMP-9 and TIMP-1 in serum Compared with the blank group, the serum MMP-9 of the model group rats increased significantly (P<0.01), and the TIMP-1 decreased significantly (P<0.01); compared with the model group, the MMP-9 of the example and comparative example groups decreased. See Table 2.

[0070]

[0071] Note: Compared with the blank group, & P<0.05, && P<0.01; compared with the model group, # P<0.05, ## P<0.01; compared with the Example 1 group, ▲ P<0.05.

[0072] Effect of Example 2 on cough and expectoration in mice 2.1 Effect on cough induced by concentrated ammonia water in mice Male Kunming mice 96, were randomly divided into 12 groups, 8 mice in each group, namely the blank control group, Bai Bei Yi Fei capsule group, examples 1-3 group and comparative examples 1-7 group, gavage drug composition 100mg / kg, respectively gavage corresponding drug composition or drug (Bai Bei Yi Fei capsule), once a day, for 7 consecutive days, the volume of drug administration 20mL / kg, 30min after the last administration, respectively, the mice were placed in 1L glass bell, through the air compressor connected to the spray head, with 400mmHg constant pressure into the bell, spray 10s, then observe the latency of cough (the time of the first cough) and the number of cough within 10min. The test results are shown in Table 3.

[0073]

[0074] Note: compared with the blank group, & P<0.05, && P<0.01; compared with the example 1 group, ▲ P<0.05.

[0075] 2.2 The expectorant effect on mice Male Kunming mice 96, were randomly divided into 12 groups, 8 mice in each group, namely the blank control group, Bai Bei Yi Fei capsule group, examples 1-3 group and comparative examples 1-7 group, gavage drug composition 100mg / kg, respectively gavage corresponding drug composition or drug (Bai Bei Yi Fei capsule), once a day, for 5 consecutive days, the volume of drug administration 20mL / kg, 30min after the last administration, respectively, intraperitoneal injection of 0.05% phenol red solution 10mL / kg, injection of phenol red 30min after the mice were killed, stripped trachea, with filter paper to absorb the blood around it. The lower edge of the tracheal cartilage to the branch was cut off, put into a test tube containing 2mL of normal saline, and then add 4% NaOH 0.1mL, the OD value was measured by CL-770 type clinical spectrophotometer at 546nm wavelength. The test results are shown in Table 4.

[0076]

[0077] Note: compared with the blank group, & P<0.05, && P<0.01; compared with the example 1 group, ▲ P<0.05.

[0078] The above detailed description is for one of the feasible embodiments of the present application, which is not used to limit the patent scope of the present application, and any equivalent implementation or change without departing from the present application shall be included in the scope of the technical scheme of the present application.

Claims

1. A method for preparing a composition for treating yin deficiency and phlegm stasis syndrome, characterized in that, The medicinal material raw material composition of the composition is: Bletilla striata, Zhejiang fritillaria, Platycodon grandiflorum, Stachys japonica, Lilium brownii, sea sponge, Aster spathulifolius, Panax notoginseng, Mahonia fortunei and licorice; The preparation method comprises the following steps: (1) Bletilla striata, Panax notoginseng, Mahonia fortunei and licorice are added with alcohol for reflux extraction to obtain alcohol extract and dregs; (2) Platycodon grandiflorum, Zhejiang fritillaria, Stachys japonica, Lilium brownii, sea sponge and Aster spathulifolius are mixed, water is added for decoction, and then microwave treatment is carried out, and filtration is carried out to obtain water extract; (3) The dregs and the water extract are mixed for enzymolysis, decoction, centrifugation, and supernatant is obtained; (4) The alcohol extract and the supernatant are mixed, concentrated, dried, and the composition is obtained.

2. The production method according to claim 1, characterized by, In step (1), the alcohol is ethanol with a volume concentration of 50%-70%; the reflux extraction is carried out at a temperature of 45-60 DEG C for 1-3h, and the solid-liquid ratio is 1:20-40g / mL.

3. The preparation method according to claim 1, characterized in that, In step (2), the solid-liquid ratio of the decoction is 1:10-25g / mL, and the decoction time is 1-3h; the microwave treatment is carried out at a power of 500-800W for 5-15min.

4. The method of claim 1, wherein, In step (3), the enzymes used for the enzymolysis are cellulase, beta-xylosidase and yeast extract enzyme, and the mass ratio of the cellulase, the beta-xylosidase and the yeast extract enzyme is 1:0.5-1:0.1-0.

3.

5. The production method according to claim 4, characterized by, The mass ratio of the cellulase, the beta-xylosidase and the yeast extract enzyme is 1:0.8:0.

2.

6. The preparation method according to claim 4, characterized in that, In step (3), the enzyme is added in an amount of 0.3%-1% of the mass of the water extract, the enzymolysis is carried out at a temperature of 50 DEG C-55 DEG C for 2h-6h, and the pH of the enzymolysis is 5.5-6.

5.

7. The preparation method according to claim 1, characterized in that, In step (3), the decoction time is 0.5-1.0h.

8. The composition for treating yin deficiency and phlegm stasis syndrome prepared by the method of any one of claims 1-7. According to weight parts, the raw material of the composition is: Bletilla striata 110-120 parts, Zhejiang fritillaria 35-45 parts, Platycodon grandiflorum 65-70 parts, Stachys japonica 65-70 parts, Lilium brownii 65-70 parts, sea sponge 65-70 parts, Aster spathulifolius 65-70 parts, Panax notoginseng 35-45 parts, Mahonia fortunei 65-70 parts, and licorice 15-25 parts.

9. The composition for treating yin deficiency and phlegm stasis syndrome prepared by the preparation method of any one of claims 1-7 is applied to the preparation of a traditional Chinese medicine preparation for treating yin deficiency and phlegm stasis syndrome.

10. Use according to claim 9, characterized in that, The dosage form of the traditional Chinese medicine preparation is selected from any one of granules, tablets, capsules, pills, powders, syrup, oral solution, oral suspension, oral emulsion, mixture and decoction.

Images