A traditional Chinese medicine composition for treating chronic bronchitis, and a preparation method and application thereof

Abstract

The present application relates to the technical field of traditional Chinese medicine formula, and particularly discloses a traditional Chinese medicine composition for treating chronic bronchitis, a preparation method and application thereof. The traditional Chinese medicine composition is composed of tremella, ganoderma lucidum, fried seminum raphani, polygala, herba epimedii, astragalus, zingiber officinale-processed magnolia officinalis, fructus schisandrae chinensis, fried pericarpium citri reticulatae, panax notoginseng, radix bis-aconiti, armeniacae semen, ginkgo seed, honey ephedra, longan arillus, eucommia ulmoides, radix morindae officinalis, radix astragali, and ginger. The traditional Chinese medicine composition can reduce the level of inflammatory factor TNF-α and increase the content of SOD according to the key points of moistening lung to relieve cough, reducing qi and resolving phlegm, and treating the symptoms and root cause, so as to reduce the oxidative damage of the body tissue and improve chronic bronchitis. The present application provides a modern pharmacological basis for treating chronic bronchitis with the traditional Chinese medicine compound.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine formulation technology, and specifically discloses a traditional Chinese medicine composition for treating chronic bronchitis, its preparation method and application. Background Technology

[0002] Chronic bronchitis (CB) is a common respiratory disease characterized by chronic nonspecific inflammation of the trachea, bronchial mucosa, and surrounding tissues. Its main clinical symptoms are cough and sputum production, and it typically lasts for several months each year for several consecutive years. Diagnosis requires further exclusion of other diseases presenting with cough, sputum production, and wheezing. The incidence of chronic bronchitis increases with age.

[0003] Current research suggests that long-term smoking is one of the main causes of chronic bronchitis. Inflammation is the core mechanism of CB development, with oxidative stress being the key component. Mucus hypersecretion, airway surface dehydration, and airway remodeling are secondary manifestations of the inflammatory response in CB. These components are not independent and promote each other. Prolonged inflammation can progress to emphysema, pulmonary hypertension, respiratory insufficiency, and even respiratory failure.

[0004] In recent years, drug treatment methods have been extensively researched and explored. Currently, the main drugs used clinically for the anti-infective treatment of chronic bronchitis are cephalosporins, fluoroquinolones, and macrolides. While there are many conventional Western medicine treatments for chronic bronchitis, long-term medication carries the risk of drug resistance. Furthermore, elderly patients are often frail and have multiple illnesses, leading to significant adverse reactions and severely impacting clinical treatment outcomes.

[0005] Traditional Chinese medicine (TCM) treatment has gained attention in recent years. The *Huangdi Neijing* (Yellow Emperor's Inner Classic) already lists cough as a disease, believing that the primary site of the lesion is the lungs, but it also involves other organs, hence the saying "all five viscera and six bowels can cause cough." The *Shanghan Lun* (Treatise on Cold Damage) and *Jinkui Yaolue* (Essential Prescriptions of the Golden Chamber) also describe cough in detail, noting that cough, wheezing, phlegm, and even chest pain, with symptoms like coughing, wheezing, and chest and back pain, can all be treated with Gualou Xiebai Baijiu Tang (Trichosanthes and Allium macrostemon Decoction with White Wine). Modern TCM clinical practice generally classifies chronic bronchitis patients into three types based on TCM syndrome differentiation: lung and spleen qi deficiency, phlegm-heat obstructing the lungs, and phlegm-turbidity obstructing the lungs. The key to treatment lies in moistening the lungs and relieving cough, regulating qi and resolving phlegm, and addressing both the root cause and the symptoms. Currently, there are many TCM formulas for treating chronic bronchitis, each with its own characteristics. However, because the diagnostic criteria for this disease vary, the treatment period is long, highly flexible, and lacks uniformity, it is difficult to achieve prevention and fundamental cure of the disease. Therefore, developing a traditional Chinese medicine composition that is versatile, clinically safe, and effective in preventing and treating symptoms during acute exacerbations and the buffer period, and comprehensively treating chronic bronchitis, has significant application value. Summary of the Invention

[0006] To address the problems existing in the prior art, this invention proposes a traditional Chinese medicine composition for treating chronic bronchitis, its preparation method, and its application. It constructs a novel traditional Chinese medicine compound and improves the compounding method of the traditional Chinese medicine composition. Different methods are used to prepare the composition based on the characteristics of the effective components of different parts, thereby obtaining a universal and effective traditional Chinese medicine composition for treating chronic bronchitis.

[0007] The technical solution adopted in this invention is as follows:

[0008] This invention provides a traditional Chinese medicine composition for treating chronic bronchitis, the composition comprising the following ingredients: Tremella fuciformis, Ganoderma lucidum, stir-fried radish seeds, Polygala tenuifolia, Epimedium, Astragalus membranaceus, ginger-processed Magnolia officinalis, Schisandra chinensis, stir-fried Citrus reticulata peel, Panax notoginseng, processed Pinellia ternata, apricot kernel, Ginkgo biloba kernel, honey-processed Ephedra sinica, longan pulp, Eucommia ulmoides, Morinda officinalis, Achyranthes bidentata, Lycium barbarum, ginseng, and ginger.

[0009] In a preferred embodiment, the traditional Chinese medicine composition comprises the following ingredients in the indicated weight ratios: 5-15g of Tremella fuciformis, 5-15g of Ganoderma lucidum, 5-15g of stir-fried radish seeds, 5-15g of Polygala tenuifolia, 5-15g of Epimedium brevicornu, 5-15g of Astragalus membranaceus, 1-10g of Magnolia officinalis (processed with ginger), 1-10g of Schisandra chinensis, 1-10g of stir-fried Citrus reticulata peel, 1-10g of Panax notoginseng, 2-8g of Pinellia ternata (processed), 1-6g of Prunus armeniaca, 1-6g of Ginkgo biloba kernel, 1-6g of Ephedra sinica (processed with honey), 1-6g of Longan pulp, 1-6g of Eucommia ulmoides, 1-6g of Morinda officinalis, 1-6g of Achyranthes bidentata, 1-6g of Lycium barbarum, 1-6g of Panax ginseng, and 0.5-4g of fresh ginger.

[0010] According to traditional Chinese medicine theory, the relevant information of the Chinese medicinal materials used in the herbal composition of this invention is as follows:

[0011] The formula contains white fungus, which is sweet and neutral in nature, and enters the lung, stomach, and kidney meridians. Its functions include nourishing and generating fluids, moistening the lungs and stomach, and tonifying lung qi. It contains white fungus polysaccharides, which have antioxidant properties, enhance cellular immunity, relieve cough and phlegm, protect liver function, and increase the activity of immune cells. It also has an adjunctive therapeutic effect on chronic bronchitis, relieving symptoms such as cough and excessive phlegm. White fungus is combined with ginkgo seeds, which astringe the lungs and relieve asthma, and is further supplemented with stir-fried dried tangerine peel to regulate qi, strengthen the spleen, dry dampness, and resolve phlegm. Together, they moisten the lungs, regulate qi, resolve phlegm, and relieve asthma. Epimedium brevicornu is then used to tonify kidney yang and strengthen muscles and bones, enhancing the overall efficacy.

[0012] The formula contains Ganoderma lucidum, which is sweet in taste and neutral in nature. It enters the heart, lung, liver and kidney meridians and has the functions of replenishing qi and calming the mind, relieving cough and asthma.

[0013] The formula includes stir-fried radish seeds, which are pungent and sweet in taste, neutral in nature, and enter the lung, stomach, and spleen meridians. The *Compendium of Materia Medica* states that radish seeds excel at regulating qi. Raw radish seeds ascend, while cooked radish seeds descend. Ascending radish seeds expel phlegm, dispel wind-cold, and promote the eruption of sores and rashes; descending radish seeds relieve phlegm, wheezing, and cough, regulate tenesmus after diarrhea, and stop internal pain—all effects of regulating qi. Raw radish seeds ascend more than descend, and can be used to treat symptoms such as food stagnation, constipation, and diarrhea due to food stagnation. Stir-fried radish seeds descend more than ascend, and can resolve phlegm and stop coughing. The stir-frying process removes toxins and warms the radish seeds, enhancing their spleen-strengthening and digestion-aiding effects. Therefore, stir-fried radish seeds are used in this formula.

[0014] The formula contains Polygala tenuifolia, which is bitter and pungent in taste, warm in nature, and enters the heart, lung, and kidney meridians. Its functions include clearing phlegm and opening the orifices, detoxifying and reducing swelling, and calming the mind and soothing the nerves.

[0015] Epimedium, a herb in the formula, is pungent and sweet in taste, warm in nature, and enters the liver and kidney meridians. Its functions include tonifying kidney yang, strengthening tendons and bones, and dispelling wind and dampness.

[0016] Astragalus in the formula is sweet and slightly warm in nature. It enters the lung and spleen meridians and has the functions of tonifying qi and raising yang, consolidating the exterior and stopping sweating, promoting diuresis and reducing swelling, generating fluids and nourishing blood, promoting circulation and relieving pain, promoting detoxification and draining pus, and promoting wound healing and tissue regeneration. It contains astragaloside and astragalin, which can reduce airway inflammation, improve asthma symptoms, inhibit tracheal smooth muscle hyperplasia, and prevent airway remodeling.

[0017] The formula contains ginger-processed magnolia bark, which is bitter and pungent in taste, warm in nature, and enters the spleen, stomach, lung, and large intestine meridians. Its functions include drying dampness and resolving phlegm, and lowering qi and relieving fullness.

[0018] Schisandra chinensis in the formula is sour and sweet in taste, warm in nature, and enters the lung, heart, and kidney meridians. Its functions are to astringe and consolidate, replenish qi and generate fluids, tonify the kidney and calm the heart. Schisandra chinensis decoction astringes the lungs and stops cough, while cough-relieving powder promotes lung qi. It is a commonly used formula for treating cough due to exogenous pathogens. In the study of the drug use patterns in the treatment of chronic bronchitis with traditional Chinese medicine, it belongs to the high-frequency drugs.

[0019] The formula contains stir-fried tangerine peel, which is bitter and pungent in taste, warm in nature, and enters the lung and spleen meridians. Its functions are to regulate qi and strengthen the spleen, dry dampness and resolve phlegm. Clinically, it is often used with prepared pinellia and ginger-processed magnolia bark to treat symptoms such as cough with excessive phlegm due to qi stagnation and dampness obstruction. It also has the value of being both a food and a medicine.

[0020] The formula contains Panax notoginseng, which is sweet and slightly bitter in taste, warm in nature, and enters the liver and stomach meridians. Its functions include dispersing blood stasis, stopping bleeding, reducing swelling, and relieving pain.

[0021] The formula contains Pinellia ternata, which is pungent and warm in nature. It enters the spleen, stomach, and lung meridians and has the functions of drying dampness and resolving phlegm, relieving nausea and vomiting, and eliminating masses and nodules.

[0022] The formula contains almonds, which are bitter, slightly warm in nature, and slightly toxic. They enter the lung and large intestine meridians and have the effects of lowering qi, relieving cough and asthma, moistening the intestines and promoting bowel movements. In clinical practice, almonds are often combined with honey-processed ephedra, ginger-processed magnolia bark, etc. to treat cough and asthma-related diseases. The almond extract amygdalin can inhibit the respiratory center, relax bronchial smooth muscle, and reduce inflammatory cell infiltration.

[0023] The ginkgo kernel in the formula is sweet, bitter and astringent in taste, neutral in nature, enters the lung meridian, and has the function of astringing the lungs and relieving asthma.

[0024] The formula contains honey-processed ephedra, which is pungent and slightly bitter in taste, warm in nature, and enters the lung and bladder meridians. Its functions include inducing sweating and dispelling cold, clearing the lungs and relieving asthma, and promoting diuresis and reducing swelling.

[0025] Longan pulp, one of the ingredients, is sweet and warm in nature, and enters the heart and spleen meridians. Its functions include nourishing the heart and spleen, replenishing blood, and calming the mind.

[0026] Eucommia bark, one of the ingredients in the formula, is sweet and warm in nature, and enters the liver and kidney meridians. Its functions include tonifying the liver and kidneys and strengthening muscles and bones.

[0027] Morinda officinalis, a component of the formula, is sweet and pungent in taste, slightly warm in nature, and enters the liver and kidney meridians. Its functions include tonifying kidney yang, strengthening tendons and bones, and dispelling wind and dampness.

[0028] The formula contains Achyranthes bidentata, which is bitter and sour in taste, neutral in nature, and enters the liver and kidney meridians. Its functions include tonifying the liver and kidneys, strengthening tendons and bones, removing blood stasis and promoting menstruation, and guiding blood downward.

[0029] The formula contains wolfberry, which is sweet in taste and neutral in nature. It enters the liver and kidney meridians and has the effects of nourishing the liver and kidneys, and benefiting essence and improving eyesight.

[0030] The ginseng in the formula is sweet and slightly bitter in taste, and slightly warm in nature. It enters the spleen, lung, heart and kidney meridians. Its functions include greatly replenishing vital energy, restoring pulse and consolidating the body, tonifying the spleen and benefiting the lungs, generating fluids and nourishing blood, and calming the mind and improving intelligence.

[0031] The formula contains ginger, which is pungent and slightly warm in nature. It enters the lung, spleen, and stomach meridians and has the functions of dispelling cold and relieving exterior symptoms, relieving nausea and vomiting, and resolving phlegm and relieving cough.

[0032] This invention relates to a novel traditional Chinese medicine composition, which is formulated to treat chronic bronchitis induced by factors such as microbial infection, dust inhalation, and inhalation of irritating gases or fumes. Combining the principles of TCM syndrome differentiation and treatment, the composition follows the treatment principle of "moistening the lungs and relieving cough, regulating qi and resolving phlegm." The formula uses Tremella fuciformis as the principal ingredient, Ginkgo biloba as the assistant ingredient, stir-fried Citrus reticulata peel as the adjuvant ingredient, and Epimedium as the guiding ingredient. The main active ingredients in the formula are those that tonify deficiency, relieve cough and asthma, resolve phlegm, relieve exterior symptoms, and astringe. They are supplemented with those that resolve dampness, regulate qi, nourish the heart and calm the mind, and promote digestion. This combination achieves the effects of clearing the lungs and relieving cough, resolving phlegm and promoting lung drainage. The treatment also takes into account both the acute exacerbation period and the recovery period. In the early stage of the disease, it is important to eliminate external pathogens, and when the pathogens are eliminated and the body is weakened, it is important to tonify the lungs and spleen.

[0033] This invention also provides a method for preparing a traditional Chinese medicine composition for treating chronic bronchitis, comprising the following steps:

[0034] (1) After pulverizing the white fungus, it was subjected to impurity removal, water extraction and alcohol precipitation to obtain the white fungus extract, and then freeze-dried to obtain the freeze-dried sample of white fungus polysaccharide.

[0035] (2) Ganoderma lucidum, stir-fried radish seeds, Polygala tenuifolia, Epimedium, Astragalus membranaceus, ginger-processed Magnolia officinalis, Schisandra chinensis, stir-fried tangerine peel, Panax notoginseng, processed Pinellia ternata, apricot kernel, ginkgo kernel, honey-processed ephedra, longan pulp, Eucommia ulmoides, Morinda officinalis, Achyranthes bidentata, wolfberry, ginseng and ginger are pulverized separately and then mixed to obtain a mixed powder;

[0036] (3) After soaking the mixed powder, it undergoes at least one or more boiling extractions, and then the water extract of the mixed raw materials is obtained after vacuum concentration.

[0037] (4) The water extract obtained in step (3) is subjected to alcohol precipitation, and the precipitate is freeze-dried to obtain the active extract of the mixed raw materials;

[0038] (5) The water extract obtained in step (3) is separated using D101 macroporous resin, eluted with ethanol-water gradient, the eluents are collected and combined, concentrated under reduced pressure, and freeze-dried to obtain a small molecule extract of the mixed raw materials.

[0039] (6) After mixing the freeze-dried sample of Tremella polysaccharide, the active extract of the mixed powder and the small molecule extract, different amounts of water were added to dilute the mixture to obtain traditional Chinese medicine compositions of different concentrations.

[0040] As a preferred embodiment, the preparation method of the Tremella fuciformis extract in step (1) is as follows:

[0041] S1-1. Mix the crushed tremella with the ethanol solution, and then remove impurities by ultrasonication and centrifugation to obtain the precipitate.

[0042] S1-2. Add the precipitate to pure water and extract it once or multiple times using a microwave extractor to obtain the extract.

[0043] S1-3. After centrifugation and drying, the extract is cooled, and the supernatant is taken and evaporated to one-third of its volume. Then it is filtered again to obtain the supernatant.

[0044] S1-4. The supernatant was subjected to alcohol precipitation to obtain a precipitate. The precipitate was purified using a DE-52 cellulose column and eluted with a gradient of sodium chloride aqueous solution. The 0.2M sodium chloride aqueous solution eluent was collected and freeze-dried to obtain a lyophilized sample of Tremella polysaccharide.

[0045] In a preferred embodiment, the solid-liquid ratio of the pulverized tremella to the ethanol solution in step S1-1 is 1g:10-30mL; the concentration of the ethanol solution is 80%.

[0046] In a preferred embodiment, the ratio of precipitate to pure water in steps S1-2 is 1:10 to 50, the output power of the microwave extractor is 400 to 500W, and the microwave extraction time is at least 2 hours.

[0047] In a preferred embodiment, the ethanol concentration in the supernatant during alcohol precipitation in steps S1-4 is 75-95%, the precipitation time is 24-30 hours, and the temperature is 4-6°C.

[0048] As a preferred embodiment, the cooking extraction in step (3) specifically involves soaking the mixed powder in 3 to 6 times the amount of water for 30 to 90 minutes, then adding water to 10 times the weight of the raw material, and decocting at least once or multiple times, each time for at least 45 minutes. The extracts are then combined and concentrated under reduced pressure to obtain the cooking liquid, with a concentration temperature of 60 to 65°C.

[0049] The present invention also applies the above-mentioned traditional Chinese medicine composition to the preparation of a drug for treating chronic bronchitis.

[0050] In a preferred embodiment, the drug is further supplemented with pharmaceutically acceptable excipients, and the dosage form is a decoction, tablet, capsule, injection, granule, powder, pill, oral liquid or syrup.

[0051] Compared with the prior art, the present invention has the following beneficial effects:

[0052] 1. The traditional Chinese medicine composition of the present invention is based on the key points of moistening the lungs and relieving cough, lowering qi and resolving phlegm, and treating both the symptoms and the root cause. It is obtained through years of experience, exploration, summarization, screening, analysis, research and verification. It has the effects of regulating qi and strengthening the spleen, clearing heat and detoxifying, moistening the lungs and relieving cough, drying dampness and resolving phlegm, removing phlegm and draining pus, dispelling wind and heat, purging the lungs and relieving asthma, promoting diuresis and reducing swelling, resolving phlegm and relieving cough, dispersing nodules and relieving pain, tonifying the spleen and replenishing qi, relieving pain and enhancing immunity. It has good efficacy, short course of treatment and no toxic side effects. It can effectively prevent and treat the symptoms of acute exacerbation and buffer period of chronic bronchitis. It has strong versatility and significant effect.

[0053] 2. Improvements are made to the compounding method of the traditional Chinese medicine composition of the present invention. Different components are prepared by different extraction methods. The compounding idea is novel. It does not affect the efficacy of the original drug. In fact, the efficacy is enhanced by changing the compounding method.

[0054] 3. The novel traditional Chinese medicine composition provided by this invention provides a modern pharmacological basis for the clinical use of this traditional Chinese medicine compound to treat chronic bronchitis. Moreover, the formula is low in cost, simple to prepare, and has broad application prospects, laying the foundation for further development of new drugs. Attached Figure Description

[0055] Figure 1 This is a pathological section (HE staining, ×100) of lung tissue from a mouse model 7 days after modeling, according to one embodiment of the present invention.

[0056] Figure 2 This is a comparison chart of cough indicators in different groups of mice according to one embodiment of the present invention;

[0057] Figure 3 This is a pathological section (HE staining, ×100) of mouse lung tissue according to one embodiment of the present invention;

[0058] Figure 4 This is a comparison chart of IL-4 levels in the serum of mice in different groups according to one embodiment of the present invention;

[0059] Figure 5 This is a comparison chart of IFN-γ levels in the serum of mice in different groups according to one embodiment of the present invention;

[0060] Figure 6 This is a comparison chart of TNF-α levels in the serum of mice in different groups according to one embodiment of the present invention;

[0061] Figure 7 This is a comparison chart of SOD content in the lung tissues of mice in different groups according to one embodiment of the present invention;

[0062] Figure 8 This is a comparison chart of IFN-γ content in the lung tissues of mice in different groups according to one embodiment of the present invention;

[0063] Figure 9 This is a comparison chart of TNF-α content in the lung tissues of mice in different groups according to one embodiment of the present invention. Detailed Implementation

[0064] The endpoints and any values ​​of the ranges disclosed herein are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of the various ranges, the endpoint values ​​of the various ranges and individual point values, and individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed herein.

[0065] The present invention will be further described below with reference to the accompanying drawings and preferred embodiments. The embodiments given are only for illustrating the present invention and are not intended to limit the scope of the present invention.

[0066] In the following examples, the terms "soaking," "water extraction," and "decoction" refer to various types of water that can be used to prepare water extracts, active extracts, and small molecule extracts, including medicinal water such as distilled water and deionized water.

[0067] In the examples described below, the alcohols used in the "alcohol precipitation" process include, for example, lower alcohols, preferably ethanol; under certain conditions, aqueous solutions of alcohols with concentrations higher than 50% v / v may also be used.

[0068] In the following embodiments, "extracting once or multiple times" and "decocting once or multiple times" mean usually extracting or decocting 1 to 3 times, preferably 1 or 2 times, and in special cases, more times can be extracted according to needs.

[0069] In the following embodiments, the experimental animals used, experimental mice, were SPF-grade, 180 male ICR mice, weighing 30 - 40 g, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., and the experimental animal production license number: SYXK(Beijing)2022 - 0052.

[0070] The Chinese medicinal materials used in the following embodiments were all purchased from the market and were qualified through identification. The experimental reagents and experimental instruments used were common experimental reagents and experimental instruments in this field, and the determination methods used were common methods in this field, except as specifically stated.

[0071] Example 1

[0072] Composition of the Chinese medicinal composition:

[0073] 7 g of Tremella fuciformis, 7 g of Ganoderma lucidum, 7 g of stir-fried Semen Raphani, 7 g of Polygala tenuifolia, 7 g of Epimedium brevicornu, 7 g of Astragalus membranaceus, 5 g of Magnolia officinalis processed with ginger, 5 g of Schisandra chinensis, 5 g of stir-fried Pericarpium Citri Reticulatae, 5 g of Panax notoginseng, 4 g of Pinellia ternata processed with method, 3 g of Armeniaca vulgaris, 3 g of Ginkgo biloba seeds, 3 g of Ephedra sinica processed with honey, 3 g of Arillus Longan, 3 g of Eucommia ulmoides, 3 g of Morinda officinalis, 3 g of Achyranthes bidentata, 3 g of Lycium barbarum, 3 g of Panax ginseng, and 2 g of Zingiber officinale.

[0074] Preparation of the Chinese medicinal composition:

[0075] First, prepare the active ingredient of Tremella polysaccharide:

[0076] (1) After pulverizing Tremella fuciformis, mix it with an ethanol solution, with a solid-liquid ratio of 1 g:20 ml, and the concentration of the ethanol solution is 80%. At the same time, perform ultrasonic impurity removal, and after impurity removal, centrifuge at 4000 rpm for 15 min to obtain a precipitate;

[0077] (2) Add the precipitate to pure water and use a microwave extractor to extract once or multiple times to obtain an extract. The material-liquid ratio of the precipitate to water is 50:1, the output power of the microwave extractor is 400 W, and the microwave extraction time is at least 2 h; after the extract is centrifuged and dried, it is cooled, the supernatant is taken and then rotary evaporated to one-third of the volume and filtered again to obtain the supernatant;

[0078] (3) Alcohol precipitation is performed on the supernatant to obtain a precipitate. The volume ratio of the supernatant to absolute ethanol is 1:4, the precipitation time is 26 h, and the temperature is 5°C; the precipitate is purified using a DE-52 cellulose column, gradient elution is performed using an aqueous sodium chloride solution, the eluate of 0.2 M aqueous sodium chloride solution is collected, and after freeze-drying, a freeze-dried sample of Tremella polysaccharide is obtained.

[0079] Secondly, prepare the active extract of the mixed powder:

[0080] (1) Ganoderma lucidum, stir-fried radish seeds, Polygala tenuifolia, Epimedium, Astragalus membranaceus, ginger-processed Magnolia officinalis, Schisandra chinensis, stir-fried Citrus reticulata peel, Panax notoginseng, processed Pinellia ternata, apricot kernel, ginkgo kernel, honey-processed Ephedra sinica, longan pulp, Eucommia ulmoides, Morinda officinalis, Achyranthes bidentata, Lycium barbarum, ginseng, and ginger are pulverized separately and then mixed to obtain a mixed powder. The powder is then passed through a 400-500 mesh sieve.

[0081] (2) After soaking the mixed powder, it undergoes at least one or more cooking extractions. The cooking extraction is specifically to soak the mixed powder in 5 times the amount of water for 60 minutes, then add water to 10 times the weight of the raw material, and cook at least once or multiple times, each time for at least 45 minutes. The extracts are combined and then concentrated under reduced pressure to obtain the cooking liquid. The concentration temperature is 60°C.

[0082] (3) The water extract obtained in step (2) is subjected to alcohol precipitation, and the precipitate is freeze-dried to obtain the active extract of the mixed raw materials.

[0083] Next, prepare the small molecule extract of the mixed powder: the water extract obtained in step (2) above is separated by D101 macroporous resin, eluted by ethanol-water gradient (0%-95%), the eluent with more than 50% ethanol is collected and combined and concentrated under reduced pressure, and then freeze-dried to obtain the small molecule extract of the mixed raw materials.

[0084] Finally, the freeze-dried sample of Tremella polysaccharide, the active extract of the mixed powder and the small molecule extract were mixed and diluted with different amounts of water to obtain traditional Chinese medicine compositions of different concentrations.

[0085] Example 2

[0086] Composition of the traditional Chinese medicine composition formula:

[0087] 15g of white fungus, 15g of Ganoderma lucidum, 15g of stir-fried radish seeds, 15g of Polygala tenuifolia, 15g of Epimedium, 15g of Astragalus membranaceus, 10g of ginger-processed Magnolia officinalis, 10g of Schisandra chinensis, 10g of stir-fried dried tangerine peel, 10g of Panax notoginseng, 10g of processed Pinellia ternata, 6g of apricot kernel, 6g of ginkgo nut kernel, 6g of honey-processed Ephedra sinica, 6g of longan pulp, 6g of Eucommia ulmoides, 6g of Morinda officinalis, 6g of Achyranthes bidentata, 6g of Lycium barbarum, 6g of ginseng, and 4g of fresh ginger.

[0088] Preparation of Traditional Chinese Medicine Compositions:

[0089] First, prepare the effective components of Tremella fuciformis polysaccharide:

[0090] (1) After crushing the white fungus, mix it with an ethanol solution with a solid-liquid ratio of 1g:20ml and an ethanol solution concentration of 80%. At the same time, remove impurities by ultrasonication. After removing impurities, centrifuge at 4000rpm for 15min to obtain the precipitate.

[0091] (2) Add the precipitate to pure water and use a microwave extractor to extract once or multiple times to obtain the extract. The ratio of precipitate to water is 50:1. The output power of the microwave extractor is 500W and the microwave extraction time is at least 2 hours. After centrifugation and drying, the extract is cooled. The supernatant is taken and evaporated to one-third of its volume, and then filtered again to obtain the supernatant.

[0092] (3) The supernatant was subjected to alcohol precipitation to obtain a precipitate. The volume ratio of supernatant to anhydrous ethanol was 1:4, the precipitation time was 30 h, and the temperature was 6℃. The precipitate was purified by DE-52 cellulose column and eluted with sodium chloride aqueous solution gradient. The 0.2M sodium chloride aqueous solution eluent was collected and freeze-dried to obtain the lyophilized sample of Tremella polysaccharide.

[0093] Secondly, prepare the active extract of the mixed powder:

[0094] (1) Ganoderma lucidum, stir-fried radish seeds, Polygala tenuifolia, Epimedium, Astragalus membranaceus, ginger-processed Magnolia officinalis, Schisandra chinensis, stir-fried tangerine peel, Panax notoginseng, processed Pinellia ternata, apricot kernel, ginkgo kernel, honey-processed Ephedra sinica, longan pulp, Eucommia ulmoides, Morinda officinalis, Achyranthes bidentata, Lycium barbarum, ginseng, and ginger are pulverized separately and then mixed to obtain a mixed powder. The powder is passed through a 400-500 mesh sieve.

[0095] (2) After soaking the mixed powder, it undergoes at least one or more cooking extractions. The cooking extraction is specifically to soak the mixed powder in 6 times the amount of water for 90 minutes, then add water to 10 times the weight of the raw material, and cook at least once or multiple times, each time for at least 45 minutes. The extracts are combined and then concentrated under reduced pressure to obtain the cooking liquid. The concentration temperature is 65°C.

[0096] (3) The water extract obtained in step (2) is subjected to alcohol precipitation, and the precipitate is freeze-dried to obtain the active extract of the mixed raw materials.

[0097] Next, prepare the small molecule extract of the mixed powder: the water extract obtained in step (2) above is separated by D101 macroporous resin, eluted by ethanol-water gradient (0%-95%), the eluent with more than 50% ethanol is collected and combined, concentrated under reduced pressure, and freeze-dried to obtain the small molecule extract of the mixed raw materials.

[0098] Finally, the freeze-dried sample of Tremella polysaccharide, the active extract of the mixed powder and the small molecule extract were mixed and diluted with different amounts of water to obtain traditional Chinese medicine compositions of different concentrations.

[0099] Example 3

[0100] Composition of the traditional Chinese medicine composition formula:

[0101] 5g of white fungus, 5g of Ganoderma lucidum, 5g of stir-fried radish seeds, 5g of Polygala tenuifolia, 5g of Epimedium, 5g of Astragalus membranaceus, 1g of ginger-processed Magnolia officinalis, 1g of Schisandra chinensis, 1g of stir-fried dried tangerine peel, 1g of Panax notoginseng, 2g of processed Pinellia ternata, 1g of apricot kernel, 1g of ginkgo nut kernel, 1g of honey-processed Ephedra sinica, 1g of longan pulp, 1g of Eucommia ulmoides, 1g of Morinda officinalis, 1g of Achyranthes bidentata, 1g of Lycium barbarum, 1g of ginseng, and 0.5g of fresh ginger.

[0102] Preparation of Traditional Chinese Medicine Compositions:

[0103] First, prepare the effective components of Tremella fuciformis polysaccharide:

[0104] (1) After crushing the white fungus, mix it with an ethanol solution with a solid-liquid ratio of 1g:20ml and an ethanol solution concentration of 80%. At the same time, remove impurities by ultrasonication. After removing impurities, centrifuge at 4000rpm for 15min to obtain the precipitate.

[0105] (2) Add the precipitate to pure water and use a microwave extractor to extract once or multiple times to obtain the extract. The ratio of precipitate to water is 50:1. The output power of the microwave extractor is 450W and the microwave extraction time is at least 2 hours. After centrifugation and drying, the extract is cooled. The supernatant is then taken and rotary evaporated to one-third of its volume and filtered again to obtain the supernatant.

[0106] (3) The supernatant was subjected to alcohol precipitation to obtain a precipitate. The volume ratio of supernatant to anhydrous ethanol was 1:4, the precipitation time was 24 h, and the temperature was 4℃. The precipitate was purified by DE-52 cellulose column and eluted with sodium chloride aqueous solution gradient. The 0.2M sodium chloride aqueous solution eluent was collected and freeze-dried to obtain the lyophilized sample of Tremella polysaccharide.

[0107] Secondly, prepare the active extract of the mixed powder:

[0108] (1) Ganoderma lucidum, stir-fried radish seeds, Polygala tenuifolia, Epimedium, Astragalus membranaceus, ginger-processed Magnolia officinalis, Schisandra chinensis, stir-fried tangerine peel, Panax notoginseng, processed Pinellia ternata, apricot kernel, ginkgo kernel, honey-processed Ephedra sinica, longan pulp, Eucommia ulmoides, Morinda officinalis, Achyranthes bidentata, Lycium barbarum, ginseng, and ginger are pulverized separately and then mixed to obtain a mixed powder;

[0109] (2) After soaking the mixed powder, it undergoes at least one or more cooking extractions. The cooking extraction is specifically to soak the mixed powder in 3 times the amount of water for 30 minutes, then add water to 10 times the weight of the raw material, and cook at least once or multiple times, each time for at least 45 minutes. The extracts are combined and then concentrated under reduced pressure to obtain the cooking liquid. The concentration temperature is 60°C.

[0110] (3) The water extract obtained in step (2) is subjected to alcohol precipitation, and the precipitate is freeze-dried to obtain the active extract of the mixed raw materials.

[0111] Next, a small molecule extract of the mixed powder is prepared: the water extract obtained in step (2) above is separated using D101 macroporous resin, eluted with an ethanol-water gradient, the eluents are collected, combined and concentrated under reduced pressure, and freeze-dried to obtain a small molecule extract of the mixed raw materials.

[0112] Finally, the freeze-dried sample of Tremella polysaccharide, the active extract of the mixed powder and the small molecule extract were mixed and diluted with different amounts of water to obtain traditional Chinese medicine compositions of different concentrations. Among them, a high-concentration traditional Chinese medicine composition solution was obtained by diluting with 10 times the amount of water for a human daily dose, and a low-concentration traditional Chinese medicine composition solution was obtained by diluting with 5 times the amount of water for a human daily dose.

[0113] Example 4

[0114] A mouse model of chronic bronchitis was constructed using LPS tracheal instillation. The therapeutic effect of the traditional Chinese medicine composition of this invention on the mouse model of chronic bronchitis was observed, and its effect on improving chronic bronchitis was determined.

[0115] In this embodiment, the traditional Chinese medicine composition prepared in Example 1 was used as the test drug (the daily adult dose is equivalent to 190g of raw herb), and ambroxol hydrochloride tablets (Shandong Yuxin Pharmaceutical Co., Ltd., batch number H20163194) were used as the positive control drug (specification: 30mg per tablet, clinical daily dose is 6 tablets).

[0116] The dosage of the traditional Chinese medicine composition of this invention is as follows: administered by gavage at a concentration of 0.1 ml / 10 g body weight. The dosages at 10 times and 5 times the concentration are 31.7 g / kg and 15.8 g / kg, respectively.

[0117] Positive control drug dosage: Dissolve ambroxol hydrochloride granules in water to a concentration of 2.25 mg / mL. The administration volume is 0.1 ml / 10 g body weight. The effective dose for mice is 0.0225 g / kg.

[0118] (1) Model preparation: Ninety 6-week-old mice were housed in an SPF-grade environment. After one week of acclimatization, 15 mice were randomly selected as the blank control group, and the remaining mice were used to prepare an animal model by intratracheal injection of lipopolysaccharide (LPS). After anesthetizing the mice with 5 mg / mL sodium pentobarbital, the mice were fixed on a mouse board, and an endotracheal tube was inserted into the trachea. LPS (concentration of 0.5 mg / mL) was slowly injected into the trachea at a rate of 0.1 mL / 10 g, and the mice were shaken up and down for 30 seconds to prevent suffocation and to help the LPS be evenly distributed in both lungs. After one week of continuous housing, three model mice were randomly selected, and hematoxylin-eosin (HE) sections were prepared from the lung specimens to assess the success of the model.

[0119] (2) Grouping and administration: After successful modeling, the model mice were randomly divided into four groups: model group, positive control group (ambroxol hydrochloride), high concentration group, and low concentration group, with 15 mice in each group. Mice in each group were administered 0.1 ml / 10 g via gavage once daily for 5 weeks. The blank control group and model group received an equal volume of water, while the intervention groups received the corresponding drugs. Samples were collected and analyzed after administration. The specific grouping and administration details for the mice are shown in Table 1 below.

[0120] Table 1. Description of drug administration in mouse groups

[0121]

[0122] (3) Measurement methods for each indicator:

[0123] Determination of cough indicators in mice: After the last administration, mice were placed in a fumigation device, smoke was injected into the fumigation device, and the latency period of the first cough and the number of coughs within 3 minutes were observed and recorded.

[0124] morphological examination of mouse lungs: Mouse lung tissue was taken, fixed in 4% paraformaldehyde solution, embedded in paraffin, sectioned, stained with hematoxylin and eosin (HE), and the pathological changes of mouse lung tissue in each group were observed under a light microscope. The light intensity of the sections was kept consistent, and the magnification was uniformly 100 times.

[0125] Determination of the levels of various inflammation-related factors in mouse serum: Blood was collected by enucleation of the eyeballs. After the blood samples were allowed to stand for 1 hour, they were centrifuged at 3000 rpm for 15 minutes. The supernatant was collected and the levels of IL-4, IFN-γ, and TNF-α in mouse serum were detected by ELISA according to the kit instructions.

[0126] Determination of the content of various related indicators in mouse lung tissue: Mouse lung tissue was taken, lung tissue homogenate was prepared, and the operation was performed according to the kit instructions to detect the content of SOD, IFN-γ, and TNF-α in mouse lung tissue.

[0127] All data were analyzed using Graphpad Prism 8.0 software, and experimental data are expressed as mean ± standard deviation (X±SD). The t-test was used to evaluate differences between groups. A p-value < 0.05 was considered statistically significant.

[0128] (4) Measurement results of each indicator

[0129] The pathological morphological changes of lung tissue sections from mice in the model group seven days after modeling are as follows: Figure 1 As shown.

[0130] The effect of the traditional Chinese medicine composition of the present invention on cough indicators in mice with chronic bronchitis is as follows: Figure 2As shown, compared with the blank control group, the latency period of the first cough in the model group was significantly different (P < 0.0001). After drug intervention, compared with the model group, the latency period of the positive control group, high concentration group, and low concentration group increased (P < 0.0001, P < 0.0001, P < 0.05). Compared with the model group, the number of coughs within 3 minutes decreased in all drug administration groups (P < 0.0001, P < 0.0001, P < 0.0001). The cough indicators in the positive control and high concentration groups were significantly improved compared with the model group.

[0131] The effects of the traditional Chinese medicine composition of the present invention on the pathological changes in lung tissue of mice with chronic bronchitis are as follows: Figure 3 As shown, in the blank control group, the bronchial walls of mice were smooth and flat, the mucosal structure was intact, no inflammatory exudate was observed in the lumen, there was no proliferation or rupture of bronchial smooth muscle, and no inflammatory cell infiltration in the lung tissue. In the model group, the bronchial mucosal epithelial cells of mice proliferated, there was a lot of inflammatory secretion in the lumen, inflammatory cell infiltration in the submucosa, alveolar dilation, and inflammatory cell infiltration. After drug intervention, the inflammatory cell infiltration in the lung tissue of mice in each treatment group was significantly improved, and the alveolar proliferation was improved. Among them, the high-concentration group and the positive control group were closer to the normal lung tissue morphology and had an improving effect on the microscopic morphology of lung tissue in mice with chronic bronchitis.

[0132] The effects of the traditional Chinese medicine composition of the present invention on the levels of various inflammatory factors in the serum of mice with chronic bronchitis are as follows: Figure 4-6 As shown;

[0133] Compared with the blank control group, the serum IL-4 level in the model group was significantly increased (P<0.0001); compared with the model group, the IL-4 levels in the positive control group, high concentration group, and low concentration group were significantly decreased (P<0.0001, P<0.0001, P<0.05).

[0134] Compared with the blank control group, the serum IFN-γ level in the model group was significantly decreased (P<0.0001); compared with the model group, the IFN-γ level in the positive control group, high concentration group, and low concentration group was significantly increased (P<0.0001, P<0.001, P<0.0001).

[0135] Compared with the blank control group, the serum TNF-α level in the model group was significantly increased (P<0.0001); compared with the model group, the TNF-α levels in the positive control group, high concentration group, and low concentration group were significantly decreased (P<0.0001, P<0.0001, P<0.001).

[0136] The effects of the traditional Chinese medicine composition of the present invention on SOD activity and IFN-γ and TNF-α content in the lung tissue of mice with chronic bronchitis are as follows: Figure 7-9 As shown;

[0137] Compared with the blank control group, the SOD content in the lung tissue of the model group was significantly decreased (P<0.001); compared with the model group, the SOD content in the positive control group, high concentration group, and low concentration group was significantly increased (P<0.01, P<0.01, P<0.05).

[0138] Compared with the blank control group, the serum IFN-γ level in the model group was significantly decreased (P<0.0001); compared with the model group, the IFN-γ level in the positive control group and the high concentration group was significantly increased (P<0.0001, P<0.01), while there was no significant difference in the IFN-γ level in the low concentration group (P>0.05).

[0139] Compared with the blank control group, the serum TNF-α level in the model group was significantly increased (P<0.0001); compared with the model group, the TNF-α levels in the positive control group, high concentration group, and low concentration group were all significantly increased (P<0.0001, P<0.0001, P<0.0001).

[0140] In summary, the components in the traditional Chinese medicine composition of this invention are well-matched, and the composition has the effect of improving chronic bronchitis. The mechanism of action may be related to regulating the balance of Th1 / Th2 cells, reducing the level of the inflammatory factor TNF-α, increasing the content of SOD, thereby reducing oxidative damage to body tissues.

[0141] The above description is merely an embodiment of the present invention and does not limit the patent scope of the present invention. Any equivalent structural or procedural transformations made based on the content of the present invention specification, or direct or indirect applications in other related technical fields, are similarly included within the patent protection scope of the present invention.

Claims

1. A traditional Chinese medicine composition for treating chronic bronchitis, characterized in that: The traditional Chinese medicine composition is made from the following raw materials in the indicated weight ratios: 5-15g of Tremella fuciformis, 5-15g of Ganoderma lucidum, 5-15g of stir-fried radish seeds, 5-15g of Polygala tenuifolia, 5-15g of Epimedium brevicornu, 5-15g of Astragalus membranaceus, 1-10g of Magnolia officinalis (processed with ginger), 1-10g of Schisandra chinensis, 1-10g of stir-fried Citrus reticulata peel, 1-10g of Panax notoginseng, 2-8g of Pinellia ternata (processed), 1-6g of apricot kernel, 1-6g of Ginkgo biloba kernel, 1-6g of honey-processed Ephedra sinica, 1-6g of longan pulp, 1-6g of Eucommia ulmoides, 1-6g of Morinda officinalis, 1-6g of Achyranthes bidentata, 1-6g of Lycium barbarum, 1-6g of ginseng, and 0.5-4g of fresh ginger.

2. A method for preparing a traditional Chinese medicine composition for treating chronic bronchitis as described in claim 1, characterized in that, Includes the following steps: (1) After pulverizing the white fungus, it is subjected to impurity removal, water extraction and alcohol precipitation to obtain white fungus extract. The liquid was then freeze-dried to obtain a freeze-dried sample of Tremella polysaccharide. (2) Ganoderma lucidum, stir-fried radish seeds, Polygala tenuifolia, Epimedium, Astragalus membranaceus, ginger-processed Magnolia officinalis, Schisandra chinensis, stir-fried tangerine peel, Panax notoginseng, processed Pinellia ternata, apricot kernel, ginkgo kernel, honey-processed Ephedra sinica, longan pulp, Eucommia ulmoides, Morinda officinalis, Achyranthes bidentata, Lycium barbarum, ginseng, and ginger are pulverized separately and then mixed to obtain a mixed powder; (3) After soaking the mixed powder, it undergoes at least two boiling extractions, and after vacuum concentration, the water extract of the mixed raw material is obtained; (4) The water extract obtained in step (3) is subjected to alcohol precipitation, and the precipitate is freeze-dried to obtain the active extract of the mixed raw materials; (5) The water extract obtained in step (3) is separated using D101 macroporous resin, eluted with ethanol-water gradient, the eluents are collected and combined, concentrated under reduced pressure, and freeze-dried to obtain a small molecule extract of the mixed raw materials. (6) After mixing the freeze-dried sample of Tremella polysaccharide, the active extract of the mixed powder and the small molecule extract, the mixture was diluted with different amounts of water to obtain traditional Chinese medicine compositions of different concentrations.

3. The method for preparing a traditional Chinese medicine composition for treating chronic bronchitis as described in claim 2, characterized in that: The preparation method of the Tremella fuciformis extract in step (1) is as follows: S1-1. Mix the crushed tremella with the ethanol solution, and then remove impurities by ultrasonication and centrifugation to obtain the precipitate. S1-2. Add the precipitate to pure water and extract it using a microwave extractor to obtain the extract; S1-3. After centrifugation and drying, the extract is cooled, and the supernatant is taken and evaporated to one-third of its volume. Then it is filtered again to obtain the supernatant. S1-4. The supernatant was subjected to alcohol precipitation to obtain a precipitate. The precipitate was purified using a DE-52 cellulose column and eluted with a gradient of sodium chloride aqueous solution. The 0.2M sodium chloride aqueous solution eluent was collected and freeze-dried to obtain a lyophilized sample of Tremella polysaccharide.

4. The method for preparing a traditional Chinese medicine composition for treating chronic bronchitis as described in claim 3, characterized in that: In step S1-1, the solid-liquid ratio of the pulverized tremella to the ethanol solution is 1g:10~30mL; the concentration of the ethanol solution is 80%.

5. The method for preparing a traditional Chinese medicine composition for treating chronic bronchitis as described in claim 3. Its features are: In steps S1-2, the ratio of precipitate to pure water is 1:10~50, the output power of the microwave extractor is 400~500W, and the microwave extraction time is at least 2 hours.

6. The method for preparing a traditional Chinese medicine composition for treating chronic bronchitis as described in claim 3, characterized in that: In steps S1-4, the supernatant undergoes alcohol precipitation with an ethanol concentration of 75-95%, a precipitation time of 24-30 hours, and a temperature of 4-6°C.

7. The method for preparing a traditional Chinese medicine composition for treating chronic bronchitis as described in claim 2, characterized in that: In step (3), the cooking extraction is specifically performed by soaking the mixed powder in 3 to 6 times the amount of water for 30 to 90 minutes, then adding water to 10 times the weight of the raw material, and decocting at least twice, each time for at least 45 minutes. The extracts are then combined and concentrated under reduced pressure to obtain the cooking liquid, with a concentration temperature of 60 to 65°C.

8. The use of the traditional Chinese medicine composition as described in claim 1 in the preparation of a medicament for treating chronic bronchitis.

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