Application of traditional Chinese medicine composition in preparation of medicine for preventing and treating bronchial asthma

Abstract

The invention relates to the technical field of traditional Chinese medicines, in particular to application of a traditional Chinese medicine composition in preparation of medicines for preventing and treating bronchial asthma. The traditional Chinese medicine composition is prepared from 5-30 parts of herba houttuyniae, 5-30 parts of honeysuckle flowers, 5-20 parts of radix paeoniae rubra, 3-15 parts of folium artemisiae argyi and 3-15 parts of herba menthae. The composition is found to have the effects of reducing the oxidative stress level, relieving the inflammation degree and relieving bronchial constriction on OVA-induced rat and mouse bronchial asthma models, and it is indicated that the traditional Chinese medicine composition has the good bronchial asthma treatment effect.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicines, and in particular to the application of a traditional Chinese medicine composition in the preparation of medicines for preventing and treating bronchial asthma. Background Art

[0002] Asthma, also known as bronchial asthma, is a heterogeneous disease characterized by chronic inflammation of the airways involving multiple cells (such as eosinophils, mast cells, T lymphocytes, neutrophils, airway epithelial cells, etc.) and cell components. This chronic inflammation is associated with airway hyperresponsiveness, and usually presents with extensive and variable reversible expiratory airflow limitation, leading to repeated attacks of wheezing, shortness of breath, chest tightness and (or) cough, etc., with intensity varying over time. It often attacks and worsens at night and (or) early morning, and most patients can relieve themselves or after treatment. If bronchial asthma is not diagnosed and treated in time, irreversible airway narrowing and airway remodeling may occur as the course of the disease prolongs. Paroxysmal expiratory dyspnea with wheezing or paroxysmal cough and chest tightness. In severe cases, patients are forced to sit or sit to breathe, have dry coughs or cough up a lot of white foamy sputum, and even develop cyanosis, etc. Sometimes coughing is the only symptom (cough variant asthma). Some adolescent patients experience chest tightness, coughing and dyspnea during exercise as the only clinical manifestations (exercise-induced asthma). Asthma symptoms can occur within minutes and can be relieved by bronchodilators or spontaneously after hours to days.

[0003] Bronchial asthma is a common chronic respiratory disease, and its treatment mainly relies on drug control. Drugs for the treatment of bronchial asthma mainly include two categories: bronchodilators and anti-inflammatory drugs. Among them:

[0004] Bronchodilators can quickly act on bronchial smooth muscles to relax them, thereby quickly relieving asthma patients' symptoms such as dyspnea, cough and chest tightness. However, bronchodilators have limited effects on long-term control of asthma inflammation and prevention of worsening of the disease. Long-term use of certain bronchodilators may cause patients to develop resistance to them, causing the drug effect to gradually weaken. Some bronchodilators may even cause side effects such as palpitations, tremors and headaches.

[0005] Anti-inflammatory drugs, especially glucocorticoids, can effectively control airway inflammation in asthma, reduce the frequency and severity of asthma attacks, and long-term use of anti-inflammatory drugs can improve lung function in asthma patients. However, long-term use of anti-inflammatory drugs may lead to side effects such as osteoporosis, increased blood sugar, and immunosuppression. In addition, inhaled glucocorticoids may also increase the risk of oral infections and Candida infections.

[0006] In addition, there are some traditional Chinese medicines that can improve the symptoms of bronchial asthma. Although these traditional Chinese medicines have certain therapeutic effects, they still have shortcomings in syndrome differentiation, drug efficacy, decoction procedures, taste, treatment courses, research support, acute phase treatment and dosage forms. Summary of the invention

[0007] In view of this, the technical problem to be solved by the present invention is to provide an application of a Chinese medicine composition in the preparation of a medicine for preventing and treating bronchial asthma.

[0008] The present invention provides the use of a Chinese medicine composition in preparing a medicine for preventing and treating bronchial asthma;

[0009] The Chinese medicine composition comprises the following raw materials in parts by weight: 5-30 parts of houttuynia cordata, 5-30 parts of honeysuckle, 5-20 parts of red peony root, 3-15 parts of wormwood leaves, and 3-15 parts of mint.

[0010] In the present invention, preventing and treating bronchial asthma includes: reducing the level of inflammatory factors, reducing the level of histamine, reducing the number of inflammatory cells and / or reducing the level of oxidative stress.

[0011] Inflammatory factors play a key role in the pathogenesis of bronchial asthma. On the one hand, they participate in the inflammatory response of the airways, leading to congestion, edema and increased secretions of the airway mucosa, further aggravating airway stenosis and dyspnea. On the other hand, inflammatory factors can also affect the contractility of airway smooth muscle and the integrity of airway epithelial cells, further aggravating the pathophysiological changes of asthma. In the present invention, the Chinese medicine composition can have a good inhibitory effect on inflammatory factors in serum and / or cytokines in alveolar lavage fluid. In some embodiments, the inflammatory factors include at least one of IL-4, IL-1β, TNFα, and IL-18.

[0012] The release of histamine is closely related to the onset of asthma. When allergens enter the body, a large amount of histamine is released and exerts its effects, thereby stimulating the contraction of bronchial smooth muscles, narrowing the bronchial cavity, and exacerbating asthma symptoms. In addition, histamine can also stimulate the secretion of mucous glands and cause inflammatory reactions. In the present invention, the composition can significantly reduce the histamine level in lung tissue.

[0013] The inflammatory response of bronchial asthma is a complex process involving multiple inflammatory cells, mediators and cytokines. The interaction between these cells and the released mediators lead to the persistence of airway inflammation and aggravate the symptoms of asthma. The experiment of the present invention shows that the composition can significantly reduce the number of inflammatory cells in alveolar lavage fluid.

[0014] Oxidative stress plays a key role in the pathogenesis of bronchial asthma. It can directly or indirectly regulate lung tissue through gene level or intracellular and extracellular signal molecules, causing a series of pathological and physiological changes such as airway mucus hypersecretion, which may eventually lead to irreversible damage to asthma. The loss or increase of intracellular or extracellular molecules related to oxidative stress signaling may jointly contribute to the occurrence and development of airway mucus hypersecretion in asthma. The experiments of the present invention show that the composition can effectively inhibit the level of oxidative stress, especially the level of oxidative stress in lung tissue.

[0015] There are many factors that lead to bronchial asthma, including but not limited to genetic factors, allergies and / or respiratory infections. In the present invention, the bronchial asthma is bronchial asthma caused by allergies. In a specific embodiment, the allergen that causes bronchial asthma is ovalbumin.

[0016] In an embodiment of the present invention, the mass fractions of each component in the traditional Chinese medicine composition are: 5-25 parts of houttuynia cordata, 5-25 parts of honeysuckle, 5-15 parts of red peony root, 3-10 parts of wormwood, and 3-10 parts of mint. Furthermore, the mass fractions of each component in the traditional Chinese medicine composition are: 15-25 parts of houttuynia cordata, 15-25 parts of honeysuckle, 10-15 parts of red peony root, 5-10 parts of wormwood, and 3-5 parts of mint.

[0017] Optionally, the mass proportions of each component in the traditional Chinese medicine composition are: 20 parts of Houttuynia cordata, 20 parts of Honeysuckle, 12 parts of Red Peony Root, 8 parts of Artemisia argyi, and 4 parts of mint.

[0018] Optionally, the mass proportions of each component in the traditional Chinese medicine composition are: 15 parts of Houttuynia cordata, 15 parts of Honeysuckle, 12 parts of Red Peony Root, 7 parts of Artemisia argyi, and 7 parts of mint.

[0019] Optionally, the mass proportions of each component in the traditional Chinese medicine composition are: 10 parts of Houttuynia cordata, 10 parts of Honeysuckle, 8 parts of Red Peony Root, 4 parts of Artemisia argyi, and 4 parts of Mint.

[0020] The composition of the present invention can be directly ground into powder, or it can be an extract obtained by conventional means in the art. The composition can be directly ground into powder by conventional methods, or it can be an extract obtained by conventional means, so as to be made into different dosage forms, obtain better therapeutic effects and be more conducive to the preparation of modern pharmaceutical dosage forms.

[0021] In an embodiment of the present invention, the preparation method of the Chinese medicine composition comprises:

[0022] Step A: weighing Chinese medicinal materials according to the weight ratio of the raw materials;

[0023] Step B: extracting honeysuckle with 6-12 times the amount of 50%-80% ethanol, filtering the extract, recovering the ethanol, drying and crushing to obtain a dry paste powder;

[0024] Step C: add 3-12 times of water to Houttuynia cordata, Artemisia argyi and mint to extract volatile oil, add coarse powder of red peony root to the residue, add 6-12 times of water to boil, filter the extract to obtain a clear paste, add ethanol with a concentration of more than 90% to 60% to 80% alcohol, precipitate with alcohol, let stand, recover ethanol from the supernatant to obtain a thick paste, dry and crush to obtain dry paste powder;

[0025] Step D: Mix the volatile oil and the two dry paste powders.

[0026] Furthermore, the method for preparing the Chinese medicine composition comprises the following steps:

[0027] Step A: Add 12 times the amount of 50% ethanol to honeysuckle, reflux extract twice, each time for 0.5 hours, filter, combine the filtrate, recover the ethanol under reduced pressure and concentrate to a thick paste with a relative density of 1.20 to 1.25, dry under reduced pressure, and crush to obtain alcohol-extracted dry extract powder;

[0028] Step B: add 8 times the amount of water to Houttuynia cordata, Artemisia argyi and mint, extract volatile oil by steam distillation for 5 hours, filter the extract and set aside;

[0029] Step C: grinding and inclusion of the obtained volatile oil with 10 times the amount of β-cyclodextrin and 20 times the amount of water for 20 minutes, drying and crushing the inclusion compound to obtain a volatile oil inclusion compound;

[0030] Step D: after extracting the volatile oil, the residue is combined with red peony root, 10 times the amount of water is added, extraction is performed twice, each time for 1 hour, the extract is combined with the oil extraction liquid, filtered, the filtrate is concentrated under reduced pressure to a clear paste with a relative density of 1.02-1.05, cooled to room temperature, 95% ethanol is added to make the alcohol content reach 70%, and the mixture is allowed to stand for more than 12 hours. The supernatant of the alcohol precipitation is reduced in pressure to recover ethanol and concentrated to a thick paste with a relative density of 1.25-1.30, dried under reduced pressure, and crushed to obtain a water-extracted dry extract powder; the alcohol-extracted dry extract powder, the volatile oil inclusion compound and the water-extracted dry extract powder are mixed.

[0031] The present invention also provides a medicine for preventing and treating bronchial asthma, which comprises pharmaceutically acceptable excipients and the following raw materials in parts by weight: 5-30 parts of houttuynia cordata, 5-30 parts of honeysuckle, 5-20 parts of red peony root, 3-15 parts of wormwood leaf, and 3-15 parts of mint.

[0032] The dosage form of the drug of the present invention includes but is not limited to decoction, granules, capsules, tablets, oral liquids, pills, tinctures, syrups, suppositories, gels, sprays and / or injections.

[0033] In some embodiments provided by the present invention, the capsule is a hard capsule or a soft capsule.

[0034] In some embodiments provided herein, the tablet is an oral tablet or an oral tablet.

[0035] Oral tablets refer to tablets for oral administration. Most of the drugs in such tablets are absorbed through the gastrointestinal tract to exert their effects, and some of the drugs in tablets exert their effects locally in the gastrointestinal tract. In some embodiments provided by the present invention, the oral tablets are ordinary compressed tablets, dispersible tablets, effervescent tablets, chewable tablets, coated tablets or sustained-release tablets.

[0036] The pharmaceutically acceptable excipients include one or a mixture of two or more of fruit powder, edible flavors, sweeteners, acidulants, fillers, lubricants, preservatives, suspending agents, edible pigments, diluents, emulsifiers, disintegrants or plasticizers.

[0037] The medicine described in the present invention also includes other medicines for preventing and treating bronchial asthma, and the other medicines for preventing and treating bronchial asthma include: at least one of budesonide, fluticasone, prednisone, dexamethasone, salmeterol, formoterol, salbutamol, terbutaline, montelukast sodium, zafirlukast, cetirizine, loratadine, aminophylline, and ipratropium bromide.

[0038] In the present invention, in the medicine, the Chinese medicine composition and other therapeutic agents may exist independently of each other or may exist in combination, and the present invention does not limit this.

[0039] Furthermore, the present invention also provides a method for preventing and treating bronchial asthma, which comprises administering the drug of the present invention.

[0040] The administration methods of the drug include, but are not limited to, oral administration, spray inhalation, rectal administration, nasal administration, buccal administration, vaginal administration, topical administration. Or non-enteral administration, such as subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intraventricular, intrasternal or intracranial injection or infusion; or administration via an explanted reservoir. Preferably, the administration method is oral administration and / or aerosol inhalation.

[0041] In the present invention, when the medicine includes a Chinese medicine composition and other therapeutic agents, two or more medicines can be administered simultaneously or sequentially, and the present invention is not limited to this.

[0042] For example, the Chinese medicine composition in the medicine is administered no longer than 6 hours, preferably no longer than 5 hours, more preferably no longer than 4 hours, even more preferably no longer than 3 hours, more preferably no longer than 2 hours and most preferably no longer than 1 hour before the administration of other therapeutic agents and / or no longer than 6 hours, preferably no longer than 5 hours, more preferably no longer than 4 hours, even more preferably no longer than 3 hours, more preferably no longer than 2 hours and most preferably no longer than 1 hour after the administration of other therapeutic agents.

[0043] In the present invention, when the medicine includes a Chinese medicine composition and other therapeutic agents, the administration frequency of two or more medicines may be the same or different, and the present invention is not limited to this.

[0044] For example, the administration frequency of two or more of the drugs is independently selected from 4 times / day, 3 times / day, 2 times / day or 1 time / day, wherein the administration frequency of the Chinese medicine composition is 4 times / day, 3 times / day, 2 times / day or 1 time / day.

[0045] In the present invention, the medicine includes a Chinese medicine composition, and the dosage is not less than the effective dose. If the medicine includes a Chinese medicine composition and other therapeutic agents, the dosage of the Chinese medicine composition is lower than the effective dose, or not lower than the effective dose; the dosage of other therapeutic agents is lower than the effective dose, or not lower than the effective dose.

[0046] In the present invention, the effective dose includes a "therapeutically effective amount" or a "prophylactically effective amount", which refers to an amount that is sufficient to treat or prevent a patient's disease but low enough to avoid serious side effects (at a reasonable benefit / risk ratio) within the scope of reasonable medical judgment. The preventive or therapeutic effective amount of a drug will vary depending on the specific drug selected (e.g., considering the efficacy, effectiveness and half-life of the drug), the selected route of administration, the disease to be prevented or treated, the severity of the disease to be prevented or treated, the age, size, weight and physical illness of the patient to be prevented or treated, the medical history of the patient to be prevented or treated, the duration of prevention or treatment, the nature of concurrent therapy, the desired preventive or therapeutic effect, and other factors, but can still be routinely determined by those skilled in the art.

[0047] In the present invention, the prevention and treatment includes prevention and / or treatment, which is a preventive and therapeutic measure for bronchial asthma. This includes a method for preventing the occurrence of bronchial asthma, the purpose of which is to prevent, slow down (reduce) undesirable physiological changes or lesions in the treated subject, and the beneficial or desired clinical results include but are not limited to avoiding the occurrence of bronchial asthma, alleviating bronchial asthma symptoms, reducing the degree of bronchial asthma, stabilizing the bronchial asthma state (i.e., not worsening), delaying or slowing the progression of bronchial asthma, improving or alleviating the bronchial asthma state, and relief (whether partial relief or complete relief), whether detectable or undetectable. Subjects in need of treatment include subjects who already have a condition or disease, as well as subjects who are susceptible to a condition or disease or subjects who intend to prevent a condition or disease.

[0048] In the present invention, the object of prevention and treatment is a human or a mammal, and the mammal includes bovines, equines, ovines, swine, canines, felines, rodents, and primates. The rodent is a mouse and / or a rat.

[0049] The Chinese medicine composition of the present invention comprises 5-30 parts of houttuynia cordata, 5-30 parts of honeysuckle, 5-20 parts of red peony root, 3-15 parts of wormwood leaf and 3-15 parts of mint. The present invention finds that the composition can reduce the level of oxidative stress, reduce the degree of inflammation and relieve bronchial contraction in OVA-induced rat and mouse bronchial asthma models, indicating that the Chinese medicine composition of the present invention has a good effect of treating bronchial asthma. DETAILED DESCRIPTION

[0050] The present invention provides the use of a Chinese medicine composition in the preparation of a drug for preventing and treating bronchial asthma. Those skilled in the art can refer to the content of this article and appropriately improve the process parameters to achieve it. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art, and they are all considered to be included in the present invention. The method and application of the present invention have been described through preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the method and application of this article without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0051] The test materials used in the present invention are all common commercial products and can be purchased in the market.

[0052] The following experiments, if no specific conditions are specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The APIs or excipients, reagents or instruments used, if the manufacturer is not specified, are all conventional products that can be purchased commercially. Unless otherwise specified, all percentages, ratios, proportions or parts are by weight.

[0053] Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein can be applied to the present invention. The present invention is further described below in conjunction with the embodiments:

[0054] Example 1 Preparation of Chinese medicine composition

[0055] Ratio of raw materials: Houttuynia cordata: 750g, honeysuckle 750g, red peony root 450g, wormwood leaf 300g, mint 150g.

[0056] Step A: adding 12 times the amount of 50% ethanol to honeysuckle, reflux extraction twice, each time for 0.5 hour, filtering, combining the filtrate, reducing pressure (-0.07MPa to -0.09MPa, 65°C to 75°C) to recover ethanol and concentrate to a thick paste with a relative density of 1.20 to 1.25 (65°C), reducing pressure drying (-0.07MPa to -0.09MPa, 60°C to 65°C), and crushing to obtain alcohol-extracted dry extract powder;

[0057] Step B: add 8 times the amount of water to Houttuynia cordata, Artemisia argyi and mint, extract volatile oil by steam distillation for 5 hours, filter the extract, store it in another container for later use;

[0058] Step C: grinding and inclusion of the obtained volatile oil with 10 times the amount of β-cyclodextrin and 20 times the amount of water for 20 minutes, drying the inclusion compound (55° C.), and crushing to obtain a volatile oil inclusion compound;

[0059] Step D: after extracting the volatile oil, the residue is combined with red peony root, 10 times the amount of water is added, and extraction is performed twice, each time for 1 hour. The extract is combined with the oil extraction liquid, filtered, and the filtrate is concentrated under reduced pressure (-0.07MPa to -0.09MPa, 75°C to 85°C) to a clear paste with a relative density of 1.02 to 1.05 (70°C), cooled to room temperature, and 95% ethanol is added to make the alcohol content reach 70%, and the mixture is allowed to stand for more than 12 hours. The supernatant of the alcohol precipitation is decompressed (-0.07MPa to -0.09MPa, 65°C to 75°C) to recover ethanol and concentrate to a thick paste with a relative density of 1.25 to 1.30 (65°C), and an appropriate amount of dextrin is added to reduce the pressure (-0.07MPa to -0.09MPa, 75°C) to dry, and pulverize to obtain a water-extracted dry extract powder.

[0060] The above three intermediates (dry extract powder of alcohol extraction part, volatile oil inclusion compound, dry extract powder of water extraction part) are mixed with appropriate amount of dextrin and stevioside, and dry granulated to prepare 1000g granules.

[0061] Example 2 Effect of Chinese medicine composition on bronchial asthma in OVA rats

[0062] 1. Experimental Materials

[0063] 1.1 Animals

[0064] 60 SD rats, male, normal grade, weighing 240-260 g.

[0065] 1.2 Drugs

[0066] A Chinese medicine composition, three intermediates prepared according to the method of Example 1, Jiangsu Kangyuan Pharmaceutical Co., Ltd.; dexamethasone sodium phosphate injection (Guizhou Guangzheng Pharmaceutical Co., Ltd., batch number 19032034); chloral hydrate (Sinopharm Chemical Reagent Co., Ltd., batch number 20200320); ovalbumin OVA (Sigma, USA, batch number SLCB8249); aluminum hydroxide (Al(OH)3) (Aladdin, batch number H2106415); physiological saline (Chenxin Pharmaceutical Co., Ltd., batch number 2101282721).

[0067] 1.3 Instruments

[0068] Electronic balance (Sartorius Scientific Instruments, model BS224S); low-temperature centrifuge (eppendorf, Germany, model 5804R); purified water instrument (Millipore, Germany, model: Milli-Q Plus); animal weight scale (Shanghai Xiangchuan Electronic Scale Co., Ltd., model: CX-SC-DA); ELISA reader (Megu Molecular, USA, model Flexstation3); cough and wheeze inducing instrument (Shanghai Xinruan, model XR-YLS-8A).

[0069] 2. Dosage design

[0070] The clinical daily dosage of the Chinese medicine composition is 72g of crude drug / day. According to the body surface area conversion method, the equivalent dose for rats is 72 / 60*6.2=7.44g of crude drug / kg, and the high, medium and low doses are 14.88, 7.44 and 3.72g of crude drug / kg respectively.

[0071] The dosage of dexamethasone sodium phosphate injection is 14.5 mg / day. According to the body surface area conversion method, the equivalent dose for rats is 14.5 / 60kg*6.2=1.5 mg / kg.

[0072] 3. Experimental Methods

[0073] The rats were fed adaptively for 3 days and had free access to feed and water. They were randomly divided into a blank control group and a modeling group. Except for the animals in the normal group, all other animals were sensitized by intraperitoneal injection of 1 mL on the first day, and repeated once on the 8th day to strengthen sensitization. At the same time, the normal group and the model group were intraperitoneally injected with an equal volume of 0.9% saline. On the 15th day, the modeling rats were placed in a cough-inducing asthma induction instrument, and 1% OVA solution was atomized and inhaled to induce asthma. The symptoms of rats showing shortness of breath, scratching the nose, sneezing, etc. were observed to be successful. The stimulation was successful once a day, 30 minutes per time, for 2 consecutive weeks. The guinea pigs with successful modeling were divided into 5 groups, including a model control group, a positive drug dexamethasone group (1.5 mg / kg), and a low, medium and high dose (14.88, 7.44, 3.72 g crude drug / kg) group of 10 each. The blank and model groups were given an equal amount of pure water, and the remaining groups were gavaged with the corresponding drug solution for 2 consecutive weeks. One hour after the last administration, the rats in each group were anesthetized with 10% chloral hydrate, blood was collected from the abdominal aorta, and bronchoalveolar lavage fluid and lung tissue were collected to detect the levels of inflammatory factors in serum and bronchoalveolar lavage fluid, and histamine in lung tissue.

[0074] 4. Statistical methods

[0075] The results were statistically analyzed using t-test.

[0076] 5. Experimental Results

[0077] 5.1 Effects of cytokines on OVA-induced asthma in rats

[0078] Compared with the normal group, the levels of TNFα and IL-4 in the serum and bronchoalveolar lavage fluid of the model group were significantly increased (P<0.001); compared with the model group, the levels of the above inflammatory factors in the serum and bronchoalveolar lavage fluid of the dexamethasone group, the high-dose group of the Chinese medicine composition, and the medium-dose group were significantly reduced (P<0.001, P<0.05). The results are shown in Tables 1 and 2.

[0079] Table 1 Effects on serum cytokines in OVA-induced asthma rat model

[0080]

[0081] Note: Compared with the normal control group, ###P<0.001; compared with the model control group, ***P<0.001, *P<0.05.

[0082] Table 2 Effects on cytokines in bronchoalveolar lavage fluid of OVA-induced asthma rat model

[0083]

[0084] Note: Compared with the normal control group, ###P<0.001; compared with the model control group, ***P<0.001, *P<0.05.

[0085] 5.2 Effects on histamine levels in OVA-induced rat lung tissue

[0086] Compared with the normal group, the histamine content in the lung tissue of the model group animals was significantly increased (P < 0.001); compared with the model group, the histamine content in the lung tissue of the rats in the dexamethasone group, the high-dose group of the Chinese medicine composition, and the medium-dose group was significantly decreased (P < 0.001, P < 0.05). The results are shown in Table 3.

[0087] Table 3 Effects on histamine levels in lung tissue of OVA-induced asthma model in rats (±S)

[0088]

[0089] 6. Experimental conclusion

[0090] The Chinese medicine composition of 14.88 and 7.44 g crude drug / kg can significantly reduce the levels of inflammatory factors in the serum and bronchoalveolar lavage fluid of rat bronchial asthma model, and can significantly reduce the content of histamine in the lung tissue of model animals, indicating that the Chinese medicine composition has a certain therapeutic effect on OVA-induced rat asthma model.

[0091] Experimental Example 3 Therapeutic effect of Chinese medicine composition on OVA-induced mouse bronchial asthma model

[0092] 1. Experimental Materials

[0093] 1.1 Animals

[0094] ICR mice, male, SPF grade, 60 mice, weighing 16-18 g, were purchased from Yangzhou University, animal certificate number: A202403070056.

[0095] Breeding environment: room temperature 20-26℃, relative humidity controlled at 40-70%, and breeding density of 5 per cage.

[0096] 1.2 Drugs

[0097] The Chinese medicine composition, the intermediate prepared according to the method of Example 1, Jiangsu Kangyuan Pharmaceutical Co., Ltd.; ovalbumin OVA (Sigma, USA, batch number SLCB8249); aluminum hydroxide (Al(OH)3) (Aladdin, batch number: H2106415); salbutamol sulfate (Jiangsu Yabang Epson Pharmaceutical Co., Ltd., batch number: 201108); sodium pentobarbital (Beijing Chemical Reagent Company, batch number: 211122); physiological saline (Chenxin Pharmaceutical Co., Ltd., batch number: 2101282721).

[0098] 1.3 Instruments

[0099] Electronic balance (Sartorius Scientific Instruments, model BS224S); low-temperature centrifuge (eppendorf, Germany, model 5804R); purified water instrument (Millipore, Germany, model: Milli-Q Plus); animal weight scale (Shanghai Xiangchuan Electronic Scale Co., Ltd., model: CX-SC-DA); multifunctional microplate reader (Megu Molecular, USA, model: Flexstation 3). Multifunctional cough and wheeze induction instrument (Shanghai Xinruan Information Technology Co., Ltd., model: XR-YLS-8A).

[0100] 2. Dosage design

[0101] The clinical daily dosage of the Chinese medicine composition is 72g of crude drug / day. According to the body surface area conversion method, the equivalent dose for mice is 72 / 60kg*12.3=14.76g of crude drug / kg, and the high, medium and low doses are 29.52, 14.76 and 7.38g of crude drug / kg respectively.

[0102] The daily dosage of salbutamol sulfate is 10 mg / day. According to the body surface area conversion method, the equivalent dose for mice is 10 mg / 60 kg*12.3=2 mg / kg.

[0103] 3. Experimental Methods

[0104] ICR mice were adaptively fed for 3 days with free access to feed and water, and were randomly divided into a normal control group (10 mice) and a model control group (50 mice) according to their body weight.

[0105] The model group animals were sensitized by intraperitoneal injection of antigen mixture (ovalbumin (OVA) 100 μg / mouse, aluminum hydroxide 2.25 mg / mouse) on the 1st and 14th days, and the normal control group mice were intraperitoneally injected with an equal volume of normal saline at the same time. Starting from the 21st day, the model group animals were given 40 mL of 1% OVA aerosol solution for sensitization and stimulation every day, and the aerosol inhalation time was 30 min / d for 7 consecutive days. The normal group mice were given an equal volume of normal saline at the same time. At the same time, the model animals were regrouped from the 21st day and divided into a positive drug group, a high-, medium-, and low-dose group of the Chinese medicine composition, with 10 mice in each group being administered. The mice in the drug administration group were gavaged with the corresponding drugs 30 minutes before aerosolization every day. The normal group and the model group were given an equal amount of normal saline once a day for 7 consecutive days.

[0106] After the last atomization, the mice were euthanized, and the bronchoalveolar lavage fluid and lung tissue were obtained. The levels of inflammatory-related factors in bronchoalveolar lavage fluid and oxidative stress indicators in lung tissue of mice were detected.

[0107] 4. Statistical methods

[0108] The results were statistically analyzed using t-test.

[0109] 5. Experimental Results

[0110] 5.1 Effects on the levels of inflammatory factors in bronchoalveolar lavage fluid of OVA-induced asthma mouse model

[0111] Compared with the normal group, the levels of inflammatory factors (IL-1β, TNFα, IL-18) and inflammatory cells (eosinophils, neutrophils, lymphocytes) in the bronchoalveolar lavage fluid of mice in the model group were significantly increased (P < 0.001). Compared with the model group, the levels of the above indicators in the salbutamol sulfate group and the high and medium dose groups of the Chinese medicine composition were significantly reduced (P < 0.01, P < 0.05). The results are shown in Tables 4 and 5.

[0112] Table 4 Effects on inflammatory factors in bronchoalveolar lavage fluid of OVA-induced asthma mouse model

[0113]

[0114] Note: Compared with the normal group, ###P<0.001; compared with the model group, **P<0.01, *P<0.05.

[0115] Table 5 Effects on inflammatory cell counts in bronchoalveolar lavage fluid of OVA-induced asthma mouse model

[0116]

[0117] Note: Compared with the normal group, ###P<0.001; compared with the model group, **P<0.01, *P<0.05.

[0118] 5.2 Effects on oxidative stress levels in OVA-induced asthma mouse model

[0119] Compared with the normal control group, the ROS and MDA activities in the lung tissue of the model group mice were significantly increased (P<0.001); compared with the model group, the activities of the above indicators in the salbutamol sulfate group and the high and medium dose groups of the Chinese medicine composition were significantly reduced (P<0.001, P<0.05). The results are shown in Table 6.

[0120] Table 6 Effects on oxidative stress levels in OVA-induced asthma mouse model

[0121]

[0122] Note: Compared with the normal group, ###P<0.001; compared with the model group, ***P<0.001, *P<0.05.

[0123] 6. Experimental conclusion

[0124] The Chinese medicine composition of 29.52 and 14.76 g crude drug / kg can significantly reduce the inflammatory response of the mouse bronchial asthma model, and significantly improve the level of oxidative stress, indicating that the Chinese medicine composition has a certain therapeutic effect on the OVA-induced bronchial asthma mouse model.

[0125] The above are only preferred embodiments of the present invention. It should be pointed out that, for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. Use of a Chinese medicine composition in the preparation of a drug for preventing and treating bronchial asthma; The Chinese medicine composition comprises the following raw materials in parts by weight: 5-30 parts of houttuynia cordata, 5-30 parts of honeysuckle, 5-20 parts of red peony root, 3-15 parts of wormwood leaves, and 3-15 parts of mint.

2. The use according to claim 1, characterized in that: The prevention and treatment of bronchial asthma includes: reducing the level of inflammatory factors, reducing the level of histamine, reducing the number of inflammatory cells and / or reducing the level of oxidative stress.

3. The use according to claim 2, characterized in that: The inflammatory factors include at least one of IL-4, IL-1β, TNFα, and IL-18.

4. The use according to claim 2, characterized in that: The inflammatory cells include at least one of eosinophils, neutrophils, and lymphocytes.

5. The use according to claim 2, characterized in that: The reducing of oxidative stress level includes reducing ROS activity and / or reducing MDA activity.

6. The use according to claim 1, characterized in that: The bronchial asthma is bronchial asthma caused by allergies.

7. The use according to any one of claims 1 to 6, characterized in that: The preparation method of the Chinese medicine composition comprises: Step A: weighing Chinese medicinal materials according to the weight ratio of the raw materials; Step B: extracting honeysuckle with 6-12 times the amount of 50%-80% ethanol, filtering the extract, recovering the ethanol, drying and crushing to obtain a dry paste powder; Step C: add 3-12 times of water to Houttuynia cordata, Artemisia argyi and mint to extract volatile oil, add coarse powder of red peony root to the residue, add 6-12 times of water to boil, filter the extract to obtain a clear paste, add ethanol with a concentration of more than 90% to 60% to 80% alcohol, precipitate with alcohol, let stand, recover ethanol from the supernatant to obtain a thick paste, dry and crush to obtain dry paste powder; Step D: Mix the volatile oil and the two dry paste powders.

8. The use according to claim 7, characterized in that: The method for preparing the Chinese medicine composition comprises the following steps: Step A: Add 12 times the amount of 50% ethanol to honeysuckle, reflux extract twice, each time for 0.5 hours, filter, combine the filtrate, recover the ethanol under reduced pressure and concentrate to a thick paste with a relative density of 1.20 to 1.25, dry under reduced pressure, and crush to obtain alcohol-extracted dry extract powder; Step B: add 8 times the amount of water to Houttuynia cordata, Artemisia argyi and mint, extract volatile oil by steam distillation for 5 hours, filter the extract and set aside; Step C: grinding and inclusion of the obtained volatile oil with 10 times the amount of β-cyclodextrin and 20 times the amount of water for 20 minutes, drying and crushing the inclusion compound to obtain a volatile oil inclusion compound; Step D: after extracting the volatile oil, the residue is combined with red peony root, 10 times the amount of water is added, extraction is performed twice, each time for 1 hour, the extract is combined with the oil extraction liquid, filtered, the filtrate is concentrated under reduced pressure to a clear paste with a relative density of 1.02-1.05, cooled to room temperature, 95% ethanol is added to make the alcohol content reach 70%, and the mixture is allowed to stand for more than 12 hours. The supernatant of the alcohol precipitation is reduced in pressure to recover ethanol and concentrated to a thick paste with a relative density of 1.25-1.30, dried under reduced pressure, and crushed to obtain a water-extracted dry extract powder; the alcohol-extracted dry extract powder, the volatile oil inclusion compound and the water-extracted dry extract powder are mixed.

9. A drug for preventing and treating bronchial asthma, comprising pharmaceutically acceptable excipients and the following raw materials in parts by weight: 5-30 parts of Houttuynia cordata, 5-30 parts of Honeysuckle, 5-20 parts of Red Peony Root, 3-15 parts of Artemisia argyi, and 3-15 parts of mint.

10. The drug according to claim 8, characterized in that Also included are other drugs for preventing and treating bronchial asthma, and the other drugs for preventing and treating bronchial asthma include: at least one of budesonide, fluticasone, prednisone, dexamethasone, salmeterol, formoterol, salbutamol, terbutaline, montelukast sodium, zafirlukast, cetirizine, loratadine, aminophylline, and ipratropium bromide.

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