Application of thermal sensitive sibistein particles in treating / or preventing asthma

Abstract

The invention discloses an application of thermal sensitive Sibestein granules in treating / or preventing asthma, and belongs to the technical field of biological medicines. The invention finds that the thermal sensitive Sibestein granules can reduce the total number of cells and the number of eosinophilic granulocytes, neutrophil granulocytes and lymphocytes in asthma mice BALF, reduce the level of cell factors in asthma mice BALF and IgE in serum, reduce exudation of inflammatory substances in airway and alveolar tissues of asthma mice and infiltration of inflammatory cells, improve the lesion degree of lung tissues, and improve the curative effect of the lung tissues. The action mechanism may be related to regulation and control of MAPK / NF-kappa B signal channel related protein expression, so that inflammatory response and cell apoptosis are inhibited, and airway inflammation of mice is relieved.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological medicine, and particularly relates to the application of hot Gansibistan granules in the treatment and / or prevention of asthma. BACKGROUND

[0002] Bronchial asthma (asthma for short) is a common chronic disease characterized by chronic airway inflammation, and 358 million people worldwide suffer from asthma. Its pathological features mainly include airway inflammation, airway hyperresponsiveness, excessive mucus secretion, and airway remodeling. The imbalance of Th1 / Th2 cells is considered to be an inducible factor of asthma. Among them, the elevated levels of Th2 cytokines, IL-4, IL-5 and IL-13, and eosinophil infiltration are closely related to persistent airway inflammation; IFN-γ is a characteristic factor of Th1 cell subsets, which inhibits the proliferation of bronchial epithelial cells and delays airway remodeling. Therefore, an effective treatment method for asthma is to attempt to improve the Th1 immune response while inhibiting the Th2 immune response to restore the Th1 / Th2 balance. Glucocorticoids are currently the most effective anti-inflammatory drugs, but have greater adverse reactions, and therefore, finding safe, effective and low-toxicity asthma prevention and treatment drugs has become a research hotspot.

[0003] Traditional Chinese medicine has a long history and profound foundation, and plays an important role in the modern medical system. Through the prevention and treatment of asthma by traditional Chinese medicine, the number of disease relapses can be reduced, the degree of relapse can be reduced, hormone dependence can be reduced, the tension and anxiety caused by asthma can be relieved, and the quality of life of patients can be improved. Hot Gansibistan granules are included in "Medicine Thought Love Azamu" and have the effect of relieving cough and reducing sputum. The prescription is composed of 12 characteristic medicinal materials such as picrotoxin, licorice, and hollyhock seeds. Because of its good curative effect and small toxic and side effects, it has been developed into a classic prescription preparation with Xinjiang characteristics, which is recorded in "Ministry of Health of the People's Republic of China Drug Standards Uygur Medicine Volume". However, the prior art has never disclosed or suggested that the drug has a specific treatment for bronchial asthma, especially through the regulation of immune balance and the inhibition of specific inflammatory pathways to improve airway inflammation. The present application uses the OVA sensitization and challenge method to construct an asthma mouse model to explore the effect of RGS on airway inflammation in asthma model mice and its possible mechanism of action, providing more data support for clinical drug use. Overcomes the shortcomings of the prior art, and for the first time discovers that hot Gansibistan granules can significantly improve airway inflammation in asthma model mice, reduce airway hyperresponsiveness, and reveal its potential mechanism of action. SUMMARY

[0004] Therefore, the present application aims to provide the application of hot Gansibistan granules in the preparation of drugs for treating and / or preventing asthma. The hot Gansibistan granules of the present application can improve airway inflammation, reduce airway hyperresponsiveness, regulate the MAPK / NF-κB signaling pathway to inhibit inflammatory response and apoptosis, and relieve airway inflammation in mice by regulating immune balance and inhibiting specific inflammatory pathways.

[0005] To achieve the above object, the present application provides the following technical solutions. The present application provides application of the thermogenic sebitan granules in preparation of a medicine for treating and / or preventing asthma.

[0006] The present application provides application of the thermogenic sebitan granules in preparation of a medicine for treating and / or preventing airway inflammation of asthma, and reducing airway hyperresponsiveness.

[0007] The present application provides application of the thermogenic sebitan granules in preparation of a medicine for treating and / or preventing MAPK / NF-κB pathway in lung tissue of asthma condition.

[0008] The present application provides application of the thermogenic sebitan granules in preparation of a medicine for treating and / or preventing airway goblet cell hyperplasia and mucus secretion of asthma condition.

[0009] The present application provides application of the thermogenic sebitan granules in preparation of a medicine for reducing Th1, Th2 cytokine and IgE level of asthma condition.

[0010] At least the following beneficial technical effects are contained: The thermogenic sebitan granules can reduce total cell number, eosinophil, neutrophil and lymphocyte number in BALF of asthma mice, reduce cytokine level in BALF and IgE level in serum of asthma mice, reduce inflammatory substance exudation and inflammatory cell infiltration in airway and alveolar tissue of asthma mice, improve lung tissue lesion degree, which may be related to regulation of MAPK / NF-κB signal pathway related protein expression, thereby inhibiting inflammatory reaction and cell apoptosis, and relieving airway inflammation of mice. BRIEF DESCRIPTION OF DRAWINGS

[0011] Figure 1 It is a diagram for observing lung tissue pathological changes of mice by HE staining.

[0012] Figure 2 It is a diagram for periodic acid-schiff (PAS) staining of lung tissue of mice in each group.

[0013] Figure 3 It is a diagram for influence of RGS on p-ERK, p-JNK, p-p38, p-NF-κB and p-IκBα protein expression in lung tissue of asthma mice; A: Control group; B: Model group; C: Experimental -L, D: Experimental -M, E: Experimental -H.

[0014] Figure 4RGS on the expression of caspase-3, cleaved caspase-3, Bax and Bcl-2 proteins in lung tissues of asthmatic mice; A: Control group; B: Model group; C: Experimental -L, D: Experimental -M, E: Experimental -H. DETAILED DESCRIPTION

[0015] The detailed description set forth below is intended as a description of various example embodiments of the application and is not intended to represent the only embodiments in which the application can be practiced. The detailed description is intended to be read with the understanding that both the comprehensive and careful presentation of the details as well as the careful presentation of the completely detailed description are not intended to limit the scope of the application but merely to illustrate certain aspects, features and embodiments of the application.

[0016] It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application. Additionally, the use of "including," "comprising," "having," "containing," and variations thereof herein is meant to encompass the items listed thereafter and equivalents thereof as well as additional items. Unless otherwise indicated, the use of the approximately or about symbol (e.g., ~) in the description or claims should not be construed to mean a deviation of 10% or less, but rather should be construed to mean a deviation of about 20%, about 10%, about 5%, about 1%, about 0.5%, about 0.1%, or the like, with the exact departure being dependent on the particular context in which the term is used.

[0017] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice of the present application, the preferred methods and materials are described herein. All documents mentioned herein are incorporated by reference to disclose and describe the methods and / or materials in connection with which the documents are cited. In case of conflict between the present specification and any document incorporated by reference, the present specification will control.

[0018] Various modifications and changes can be made to the specific embodiments of the application described herein without departing from the scope or spirit of the application. Other embodiments of the application will be apparent to those of ordinary skill in the art from the description and examples presented herein. The specification and examples given herein are exemplary and should not be used to limit the present application, unless otherwise specifically indicated.

[0019] As used herein, the terms "comprises", "comprising", "includes", "including", "has", "having", "contains", "containing", or variations thereof, are intended to mean a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises, includes, has, or contains a list of elements does not include only those elements but can also include other elements not expressly listed or inherent to such process, method, article, or apparatus.

[0020] As used herein, the terms "room temperature", "ambient temperature" are intended to mean 25 ± 2 °C, unless otherwise specified.

[0021] The raw materials and instruments used in the following examples of the present application are commercially available, unless otherwise specified.

[0022] Examples 1. Experimental materials Animals: SPF grade BALB / c mice (female), 6-8 weeks of age, weighing 18-20 g, purchased from the Animal Experiment Center of Xinjiang Medical University. License No.: SCXK (Xin) 2023-0002. The mice were raised in a clean animal laboratory with day and night alternation, the feeding temperature was 18-22℃, and the mice were free to eat and drink water, and were adaptively fed for 1 week. The animal experiment was approved by the Animal Ethics Review Committee of Xinjiang Uygur Autonomous Region Institute of Pharmaceutical Research (Ethical Approval No. XJIMM-20240604).

[0023] Drugs and reagents: Gansensibistan granules were provided by Xinjiang Hetian Uygur Pharmaceutical Co., Ltd.; dexamethasone acetate tablets (batch number: H50020446, Chongqing Kerui Pharmaceutical Group Co., Ltd.); Feinan tablets (batch number: Z20080609, Jilin Zhenghe Pharmaceutical Group Co., Ltd.); OVA (purity: ≥98%), aluminum hydroxide gel and normal saline were purchased from Jilin Zhengye Biological Products Co., Ltd.; interferon-γ (IFN-γ) was purchased from Wuhan Dr. Deki Biological Engineering Co., Ltd.; tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), IL-4, IL-5, IL-13, immunoglobulin E (IgE) were purchased from Wuhan Elireite Biological Technology Co., Ltd.; Periodic acid-Schiff (PAS) staining kit (purchased from Beijing Solabio Technology Co., Ltd.); phosphorylation-extracellular signal-regulated kinase 1 / 2 (p-Erk1 / 2) antibody, Erk1 / 2 antibody, phosphorylation-c-Jun-terminal kinase (p-JNK) antibody, JNK antibody, phosphorylation-mitogen activated protein kinase (p-MAPK)-p-p38MAPK antibody, p38MAPK antibody, rabbit anti-mouse B cell lymphoma-2 (Bcl-2) antibody, Bcl-2 associated X protein (Bax) antibody, phosphorylation-nuclear factor kappa-B (p-NF-κB) antibody, NF-κB antibody, phosphorylation-inhibitor alpha of NF-κB (p-IκB-α) antibody were purchased from Cell Signaling Technology Co., Ltd.; caspase-3, cleaved-caspase-3 antibody were purchased from Abeam; mouse anti-β-tubulin antibody was purchased from Beijing Zhongsu Jinqiao Biological Technology Co., Ltd.; goat anti-mouse IgG / horseradish peroxidase-labeled, goat anti-rabbit IgG / horseradish peroxidase-labeled were purchased from Affinity, USA; Bicinchoninic acid (BCA) protein quantification kit was purchased from Thermo Fisher Scientific, USA.

[0024] Instruments: Multifunctional microplate detector (TECAN SPARK, Switzerland); ultrasonic cell crusher (Ningbo Xinzhi Biological Technology Co., Ltd.); electric heating constant temperature air drying oven (Shanghai Qixin Scientific Instrument Co., Ltd.); multifunctional imaging system (VILBER, France); electrophoresis apparatus and electrophoresis tank (Bio-Rad, USA); table type high speed refrigerated centrifuge TGL-16k (Hunan Xiangyi Laboratory Instrument Development Co., Ltd.); optical microscope (OLYMPUS, Japan); ice maker (Hangzhou Zhongleng Electrical Co., Ltd.); vortex shaker (Jiangsu Haimen Qilunbel Instrument Manufacturing Co., Ltd.); micropipette (Eppendorf AG, Germany).

[0025] 2. Test method 2.1 Establishment of animal model and drug administration The mice were randomly divided into a blank group, a model group, RGS low, medium and high dose groups (0.75, 1.5, 3 g•kg -1 ), a dexamethasone positive drug group (DEX, 3 mg•kg -1 ), and a Fei Ning tablet positive drug group (FN, 0.72 g•kg -1 ). Each group of 6 mice was adaptively fed for 1 week. Except for the blank group, the remaining mice were given OVA sensitizing solution 200 μL (containing OVA 20 μg) intraperitoneally on the 1st, 8th and 15th days. From the 22nd to the 28th day, except for the blank group, each drug administration group was administered according to the set dose. One hour after gavage, except for the normal control group, the mice in each drug administration group were placed in the atomizer in turn according to the group, and were atomized with 1% OVA normal saline solution. The normal group was atomized with normal saline, once a day, 30 min each time, for 7 days.

[0026] 2.2 Specimen collection 24 h after the last treatment, the mice were anesthetized with sodium pentobarbital, and one side of the mouse eyeball was removed with an ophthalmic forceps. The mouse blood was collected in a 1.5 ml centrifuge tube, which was left to stand at room temperature and then centrifuged. The supernatant was stored in a freezer at -80 ℃. After being sacrificed, the trachea was cannulated and 800 μL of ice-cold phosphate buffered saline (PBS) was slowly injected into the lungs. After repeating the process once, the BALF was collected. The mouse lung tissue was separated, the left lung was placed in paraformaldehyde for subsequent sectioning, and the right lung was stored in a freezer at -80 ℃ for subsequent experiments.

[0027] 2.3 Classification and counting of inflammatory cells in BALF The alveolar lavage fluid was centrifuged at 1000× r•10 min at 4 ℃, the precipitated cells were resuspended in 200 μL PBS, and the total number of cells, eosinophils (EOS), neutrophils (Neu) and lymphocytes (Lym) were counted using a Mindray automatic blood cell analyzer.

[0028] 2.4 Detection of active factors in serum and BALF by ELISA method The serum was taken and the content of IgE was determined by ELISA method. The BALF supernatant was taken and the contents of INF-γ, IL-4, IL-5, IL-13, IL-1β and TNF-α were determined by ELISA method.

[0029] 2.5 Pathological changes of lung tissue in asthmatic mice The lung tissue of mice was fixed with paraformaldehyde, embedded in paraffin, cut into 4 μm sections, stained with hematoxylin-eosin (HE), and observed under a microscope for inflammatory cell infiltration, alveolar, mucosal and epithelial cell damage. PAS staining was used to observe the mucus secretion of goblet cells.

[0030] 2.6 Detection of protein expression by Western Blot method The protein of the right lung tissue of mice was extracted, RIPA cell lysis solution was added, and lysis was performed on ice for 30 min. The supernatant was collected by centrifugation, the cell protein was extracted, the protein concentration was detected using BCA kit, and the protein concentration of each group was diluted and adjusted to the same level. After boiling in a 100 ℃ metal bath for 5 min, it was stored in aliquots. The protein samples were subjected to polyacrylamide gel electrophoresis at 10 % and then transferred to PVDF membrane. 5 % skim milk powder was used to block the membrane at room temperature for 2 h, and TBST was used to wash 3 times for 10 min each time. The corresponding primary antibody was added and incubated at 4 ℃ overnight. The next day, TBST was used to wash 3 times, and the corresponding secondary antibody was added and incubated at room temperature for 2 h. TBST was used to wash 3 times, and chemiluminescence reagent was added. The gel imaging system was used to take pictures and Image J software was used to analyze the gray scale of the bands.

[0031] 2.7 Statistical analysis The experimental data were statistically analyzed using GraphPad Prism 9.5 statistical software. All data were expressed as The sample means were compared using one-way ANOVA, and P<0.05 was considered statistically significant.

[0032] 3. Experimental results 3.1 Effect of RGS on the classification and counting of inflammatory cells in BALF of mice Compared with the model group, the total number of cells and the number of each type of cell (such as eosinophils (EOS), neutrophils (Neu), and lymphocytes (Lym)) in the BALF of the mice in the model group increased significantly, and the differences were statistically significant (all P<0.01); compared with the model group, the total number of cells, the number of eosinophils, the number of neutrophils, and the number of lymphocytes in the mice in the low-, medium-, and high-dose experimental groups decreased, and the differences were statistically significant (all P <0.05 or P<0.01). See Table 1.

[0033] Table 1 Statistical table of cell classification count in the bronchoalveolar lavage fluid (BALF) )

[0034] EOS: eosinophils; Neu: neutrophils; Lym: lymphocytes; compared with the normal group, * P< 0.05, ** P< 0.01; compared with the model group, # P< 0.05, ## P< 0.01. 3.2 RGS on the secretion of Th1 and Th2 type cytokines in the BALF of mice and IgE in serum Since Th1 / Th2 cytokines are crucial in asthma, imbalance of these cytokines can lead to allergic asthma. Compared with the normal group, the levels of IL-4, IL-5, and IL-13 in the BALF of the mice in the model group increased significantly, while the level of INF-γ decreased, and the differences were statistically significant (all P<0.01); compared with the model group, the levels of IL-4, IL-5, and IL-13 in the BALF of the mice in the low-, medium-, and high-dose experimental groups decreased, while the level of INF-γ increased, and the differences were statistically significant (all P<0.01). In addition, the level of IgE in serum was detected by ELISA. Compared with the normal group, the level of IgE in serum in the model group increased; compared with the model group, the level of IgE in serum in the low-, medium-, and high-dose experimental groups decreased after treatment. The above results show that the RGS granules can reduce Th1 and Th2 cytokines and IgE levels to alleviate OVA-induced asthma, and the differences were statistically significant (all P <0.05 or P<0.01). See Table 2.

[0035] Table 2 Effect of RGS on the secretion of Th1 and Th2 cytokines in the BALF and IgE in serum )

[0036] Doses were consistent with Table 1; IL-4: interleukin-4; IL-5: interleukin-5; IL-13: interleukin-13; INF-γ: interferon-γ; IgE: immunoglobulin E; *P<0.05 compared with the normal group, ** P< 0.01; compared with the model group, # P<0.05, ## P<0.01. 3.3 Effects of RGS on secretion of inflammatory factors in BALF of mice Compared with the blank group, the levels of IL-1β and TNF-α in BALF of the model group were increased, and the differences were statistically significant (both P<0.01); compared with the model group, the levels of IL-1β and TNF-α in BALF of the low-, medium-, and high-dose experimental groups were decreased, and the differences were statistically significant (both P<0.01); compared with the normal group, the levels of IL-1β and TNF-α in BALF of the low-, medium-, and high-dose experimental groups were decreased, and the differences were statistically significant (both P<0.01). P <0.05 or P<0.01). See Table 3.

[0037] Table 3 Effects of RGS on secretion of inflammatory factors in BALF )

[0038] Doses were consistent with Table 1; IL-1β: interleukin-1β; TNF-α: tumor necrosis factor-α; *P<0.05 compared with the normal group, * P< 0.05, **P<0.01; compared with the model group, # P< 0.05, ## P< 0.01 3.4 Effects of RGS on inflammatory cell infiltration and mucus secretion in lung tissues of mice The results of HE staining showed that, compared with the blank group of mice, inflammatory cells of the asthma group of mice were widely infiltrated around the trachea and blood vessels, and RGS or DEX treatment could significantly improve this condition. See Figure 1 The results of PAS staining showed that, compared with the blank group of mice, the model group of mice had obvious hyperplasia of goblet cells around the airway and increased mucus secretion, and RGS or DEX treatment could significantly inhibit the hyperplasia of goblet cells and mucus secretion in the airway of the asthma mice. See Figure 2 .

[0039] 3.5 Effects of RGS on expression of MAPK / NF-κB pathway-related proteins in lung tissues of mice The p-ERK / ERK expression of the blank group, model group and low, medium and high dose experimental groups was 0.39±0.12, 1.14±0.09, 1.13±0.17, 0.86±0.01 and 0.43±0.1 respectively, the p-JNK / JNK expression was 0.40±0.05, 0.94±0.09, 0.70±0.09, 0.54±0.08 and 0.5±0.1 respectively, the p-p38 / p38 expression was 0.46±0.04, 0.98±0.12, 0.90±0.09, 0.69±0.10 and 0.47±0.04 respectively, the p-NF-κB / NF-κB expression was 0.81±0.14, 1.26±0.13, 0.91±0.07, 0.82±0.20 and 0.72±0.02 respectively, and the p-IκBα / IκBα expression was 0.46±0.14, 1.28±0.13, 0.76±0.09, 0.65±0.13 and 0.68±0.18 respectively. The p-p38 / p38, p-ERK / ERK, p-JNK / JNK, p-NF-κB / NF-κB and p-IκBα / IκBα expressions of the model group were significantly higher than those of the normal group, and the difference was statistically significant (P<0.05 or P<0.01); the p-p38 / p38, p-ERK / ERK, p-JNK / JNK, p-NF-κB / NF-κB and p-IκBα / IκBα expressions of the low, medium and high dose experimental groups were lower than those of the model group (P<0.05 or P<0.01). See Table 3. P <0.05 or P<0.01); the p-p38 / p38, p-ERK / ERK, p-JNK / JNK, p-NF-κB / NF-κB and p-IκBα / IκBα expressions of the low, medium and high dose experimental groups were lower than those of the model group (P<0.05 or P<0.01). See Table 3. P <0.05 or P<0.01). See Table 3. Figure 3 .

[0040] 3.6 Effect of RGS on the expression of apoptosis-related proteins in mouse lung tissue The caspase-3 expression of the blank group, model group and low, medium and high dose experimental groups was 0.73±0.11, 0.81±0.03, 0.91±0.04, 0.82±0.02 and 0.78±0.06 respectively, the cleaved-caspase-3 / caspase-3 expression was 0.46±0.03, 1.09±0.07, 0.68±0.12, 0.84±0.15 and 0.76±0.10 respectively, the Bax expression was 0.65±0.09, 0.97±0.11, 0.75±0.04, 0.81±0.15 and 0.69±0.05 respectively, and the Bcl-2 expression was 0.86±0.09, 0.44±0.11, 0.47±0.14, 0.95±0.01 and 0.80±0.05 respectively. The cleaved-caspase-3 / caspase-3 and Bax expressions of the model group were significantly higher than those of the normal group, and the Bcl-2 expression was significantly lower, and the difference was statistically significant (P<0.05 or P<0.01); the cleaved-caspase-3 / caspase-3, Bax and Bcl-2 expressions of the low, medium and high dose experimental groups were lower than those of the model group (P<0.05 or P<0.01). See Table 4. P<0.05 or P<0.01); the expression of cleaved-caspase-3 / caspase-3 and Bax in the low, medium and high dose experimental groups was significantly lower than that in the model group, and the expression of Bcl-2 was significantly higher, and the differences were statistically significant (P<0.05 or P<0.01). P <0.05 or P<0.01). See Figure 4 .

[0041] In summary, the thermosensitive saibian granules have good anti-inflammatory activity. After the intervention of the thermosensitive saibian granules in the present application, the total number of cells, eosinophils, neutrophils and lymphocytes in the BALF of the mice are significantly reduced; the bronchial wall thickening, lung tissue and peribronchial inflammatory cell infiltration of the mice are reduced; the bronchial wall thickening, lung tissue and peribronchial inflammatory cell infiltration of the mice are reduced; and the levels of IL-4, IL-5, IL-13, TNF-α and IL-1β in the BALF of the mice and the level of IgE in the serum of the mice are significantly reduced. It is shown that RGS can reduce the release of inflammatory factors and effectively relieve the airway inflammation of the asthmatic mice.

[0042] Helper T cells play a core role in the immune system, including Th1 and Th2 subgroups, and the imbalance of Th1 / Th2 response is an important mechanism leading to and exacerbating allergic asthma. IL-13, IL-4 and IL-5 are Th2 type cytokines, mainly involved in mucus secretion, eosinophil production, IgE synthesis and airway hyperresponsiveness; compared with Th2 cells, Th1 cells have opposite effects. IFN-γ is a Th1 cell type factor, and its main function is to inhibit the synthesis of IgE and the differentiation of Th2 type cytokines. Th1 cells can inhibit the differentiation and proliferation of eosinophils and basophils, and their activity is controlled by Th2 cell synthesized cytokines. Therefore, an effective treatment method for asthma is to improve Th1 immune response while inhibiting Th2 immune response to restore Th1 / Th2 balance. The results of the present application show that the levels of Th2 type cytokines (IL-13, IL-4 and IL-5) in the BALF of the asthmatic mice are decreased and the level of Th1 cytokine (IFN-γ) is increased after the intervention of RGS. The results show that RGS can promote Th1 / Th2 balance to inhibit allergic asthma, and these results show that RGS can be a potential anti-inflammatory agent for treating asthma.

[0043] On this basis, the present application further studies the lung tissue to determine the specific mechanism of RGS in improving asthma inflammation. Experiments show that MAPK / NF-κB is proved to be an important signaling pathway in the onset of asthma. MAPK / NF-κB signaling pathway is involved in airway inflammation, airway remodeling and apoptosis. The MAPK family is composed of extracellular signal-regulated kinase (ERK), c-Jun amino-terminal kinase (JNK) and p38 signaling pathway. ERK regulates the biosynthesis of pro-inflammatory and anti-inflammatory cytokines to participate in airway inflammation, and is involved in IL-5-mediated eosinophilia and IL-13-dependent mucus hypersecretion. The activation of JNK is related to IgE class switching and is involved in airway remodeling. Activated p38MAPK induces polarization of Th2 cells and production of Th2 cytokines, inhibits eosinophil apoptosis, and produces inflammatory factors, thereby promoting the formation of asthma inflammation. The specific substrate after p38MAPK phosphorylation can activate the NF-κB pathway, further promote the expression of inflammatory factors, and the secretion of Th2 cytokines leads to the overreaction of the body's immune system. NF-κB is combined with inhibitory proteins in the form of p50-p65 heterodimer, is activated and induces gene transcription when the cell is stimulated, regulates the abnormal expression of downstream cytokines such as TNF-α, IL-6 and IL-1β, and further expands and aggravates the inflammatory response of asthma. The results of the present application show that the phosphorylation of p38MAPK, JNK, ERK, NF-κB and IκBα in the lung tissue of the asthma model mice increases to different degrees, while the phosphorylation of p38MAPK, JNK and ERK decreases after the intervention of RGS. These results show that RGS can alleviate the airway inflammatory response of mice and relieve asthma through the MAPK / NF-κB pathway.

[0044] Apoptosis is a kind of programmed cell death, which plays an important role in cell growth, morphogenesis, tissue homeostasis and immunity, etc. Bax is a protein that promotes tumor cell apoptosis, which transfers to the mitochondrial membrane in monomer form, and undergoes conformational change, mediates the release of downstream apoptosis molecules, thereby triggering cell apoptosis. Bcl-2 can maintain normal cells and prevent apoptosis by mediating the above pathways. Caspase-3 is a very important protein molecule in the apoptosis pathway, and various apoptosis pathways induce cell apoptosis through caspase-3 signaling. Experiments show that under normal physiological conditions, the renewal metabolism of airway epithelial tissue is in a dynamic balance state under the action of pro-apoptotic and anti-apoptotic factors, but when in the pathogenesis of asthma, due to the long-term repeated stimulation of chronic inflammation, the expression of pro-apoptotic genes decreases, and the expression of anti-apoptotic genes increases, so that the airway tissue appears hyperplasia, proliferation and other airway remodeling changes. The results of the present application show that RGS can down-regulate the expression levels of Bax, cleaved caspase-3 and protein in the lung tissue of asthmatic mice, and up-regulate the expression level of Bcl-2 protein, thereby improving airway inflammation and relieving the development of asthma.

[0045] In summary, RGS can reduce the total number of cells, eosinophils, neutrophils and lymphocytes in the BALF of asthmatic mice, reduce the levels of cytokines in the BALF and IgE in the serum of asthmatic mice, reduce the exudation of inflammatory substances and the infiltration of inflammatory cells in the airway and alveolar tissue of asthmatic mice, improve the degree of lung tissue lesions, and may be related to the regulation of MAPK / NF-κB signaling pathway-related protein expression, thereby inhibiting inflammatory response and apoptosis and relieving airway inflammation in mice.

[0046] The above only describes the preferred embodiments of the present application, and it should be noted that those skilled in the art can make several improvements and refinements without departing from the principles of the present application, and these improvements and refinements should also be considered within the protection scope of the present application.

Claims

1. Application of thermosensitive bistan granules in the preparation of drugs for the treatment and / or prevention of asthma.

2. Application of thermosensitive Sepitan granules in the preparation of drugs that inhibit the MAPK / NF-κB pathway in lung tissue of asthmatic patients.

3. Application of thermosensitive Sepitan granules in the preparation of drugs that inhibit airway goblet cell proliferation and mucus secretion in asthma.

4. Application of thermosensitive cytokines in the preparation of drugs that reduce the levels of Th1, Th2 cytokines and IgE in asthma.

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