Professor Miao Qing's Experience In Treating Cough Variant Asthma And The Mechanism Of Jinbozhiketang Regulating TRPV1

Cough variant asthma is asthma with chronic cough as the primary manifestation,which can be induced or aggravated by factors such as speech,dust,and irritating odor.It is often manifested by the high airway reactivity,rather than symptoms such as fine shortness of breath.Modern medicine mainly uses glucocorticoids,bronchodilators,and anti-inflammatory treatment.However,some patients have poor treatment results that are failing to improve airway sensitivity.The long-term coughing affects the work and life of CVA patients and increases the psychological burden of patients.Prof.Miao Qing has been engaged in TCM clinical,teaching,and scientific research for many years.With extensive experience in the treatment of CVA in TCM,he developed the theory of evidence-based treatment of CVA based on external factors and internal disharmony that has remarkable clinical efficacy.Under the tutor's guidance,we summarized his experience in treating CVA from the perspective of external factors and internal disharmony.We chose his common prescription that is called Jin Fei Zhi Ke soup,based on the external factors,to carry out research of animal experiment.The research is to explore the mechanism of CVA and TRPV1,and the efficacy of Jin Fei Zhi Ke soup in treating CVA rats from the perspective of transient receptor potential vanilloid 1(TRPV1).Part ? Literature ReviewThis paper collects the records of ancient Chinese medicine in the typical ancient books on the principle-method-recipe-medicines of long-time cough.We bring the thought and experience of the famous modern Chinese medicine experts in treating long-time cough together and understand thoroughly.And we discuss the understanding and treatment of contemporary Chinese medicine practitioners in treating CVA by categorizing it into wind cough,cold cough,dry cough,phlegm stasis cough,liver cough,spleen cough,and kidney cough.Part ? Summary of Professor Miao Qing's experience in cough variant asthma Purpose;By collecting the experience of Prof.Miao Qing in treating CVA from the perspective of external and internal factors,to gain an in-depth understanding of the mentor's thoughts and methods of identifying and treating CVA from the perspective of internal and external pathogenic factor.Method:Through outpatient and ward study,collating clinical cases,lessons by the instructor,and the combination of ancient books and modern literature research,we summarized the instructor's thoughts and methods of treating CVA in terms of external factors and internal disharmony,and the law of medication addition and subtraction.We also provided case studies to support.Result;The instructor believes that CVA is caused by the wind,cold,damp and dryness in the case of externally factors,and it can also be caused by internal disharmony in the case of liver,spleen and kidney dysfunction and internal turbidity in the lung.If caused by wind,we use modified Zhisou powder to dispel the wind and cease the cough.If caused by cold,we use modified Jinfeicao powder and Zhisou powder,to warm the lung and cease the cough.If caused by damp,we use modified Ma Xing Yi Gan soup to dissipating dampness and cease the cough.If caused by dryness,we use Mai Men Dong soup or Qing Zao Jiu Fei soup to dispel the wind,moisten the lung and cease the cough.If caused by the conflict between lung and liver,we use Xiao Chai Hu soup with Zhisou powder to smooth the liver and cease the cough.If caused by spleen deficiency,we use Si Jun Zi soup to tonify the spleen,reduce the phlegm and cease the cough.If caused by the kidney deficiency,we use Er Xian soup to warm the kidney and cease the cough.Conclusion:Prof.Miao Qing thought the treatment of CVA should be based on two factors:external factors and internal disharmony.We must focus on the origin of CVA.The main principle of curing CVA is dispelling illness and ceasing the cough with different targets to internal or external pathogenic factors.Part ?:Mechanism of regulation of TRPV1 by Jin Fei Zhi Ke soupPurpose:To explore the mechanism of TRPV1 in the pathogenesis of CVA,as well as the efficacy and mechanism of Jin Fei Zhi Ke soup in treating CVA rats through animal experiments of Jin Fei Zhi Ke soup regulating TRPV1.Method:126 healthy male rats were divided into 7 groups by random rule,including blank control group,model group,glucocorticoid group,herbal medicine low,medium and high dose group and TRPV1 antagonist group.Except for the blank group,the remaining groups were modeled with reference to the literature:on day 1,each rat was given a solution of ovalbumin and aluminum hydroxide at 10 points in the hind-plantar,inguinal,lumbar,dorsal and cervical regions of the rats,respectively;0.05ml was injected subcutaneously at each point and 0.5ml was injected intraperitoneally for a total of 1ml;on day 8(1 week later),the rats were repeated once to enhance sensitization;from day 15,the rats were placed in a homemade transparent nebulized glass box(50cm×50cm×30cm)and given 1%(10mg/ml)ovalbumin solution for 20min to excite CVA for 10 days.At 3 hours,2 hours and 1 hour before the detection of various indicators,the TRPV1 antagonist group was intervened with 10?mol/L Capsazepine solution atomized for 10 minutes;the rest of the administration group started intragastric administration for 28 consecutive days after the last ovalbumin solution atomization challenge.After reaching the dosing cycle,airway resistance was measured to obtain airway resistance values.Enzyme-linked immunosorbent assay(ELISA)was used to detect eosinophilic cationie protein(ECP),interleukin-4(IL-4),interleukin-5(IL-5),transforming growth factor-?1(TGF-?1)of serum and bronchoalveolar lavage fluid,hematoxylin-eosin staining(HE staining)was used to stain the trachea and lung tissue of rats,and the pathological morphological changes were observed under the microscope;Western blotting was used to detect TRPV1 protein expression in each group.Result:(1)Pathological observations of the main bronchial tubes and lung tissues of rats in each group:the epithelium of the main bronchial mucosa of the blank control rats was continuously intact,with basically no shedding cells in the lumen and no inflammatory cell infiltration in the tube wall and surrounding tissues;the structure of the fine bronchial tubes and alveoli was clear and intact.And the epithelium of the fine bronchial mucosa was intact,with basically no shedding cells and no obvious inflammatory cell infiltration around the tube.In the model group,the epithelium of the main bronchial mucosa of rats was discontinuous and incomplete,the epithelium of the bronchial mucosa was dislodged,and inflammatory cells were infiltrated in the tube wall and surrounding tissues;a large number of inflammatory cells were infiltrated around the wall of the fine bronchial tubes,visible secretions in the tube lumen,alveolar spacing was widened,and some alveoli fused.The epithelium of the main bronchial mucosa of the rats in each administration group was intact,and the degree of epithelial shedding and inflammatory cell infiltration of the bronchial mucosa and the surrounding tissues were reduced to a different extent than that of the CVA model group;the lumen and alveolar structure of the fine bronchial tubes were intact,and the degree of epithelial shedding and inflammatory cell infiltration of the airway were reduced to a different extent than that of the model group.(2)Behavioral characteristics of rats in each group:no special behavioral abnormalities in the blank control group.Rats in the model group showed behaviors such as faster breathing and slight twitching of the abdominal limbs after sensitization to ovalbumin.Rats after stimulated by the ovalbumin nebulization showed faster breathing,scratching of the nose,urination and defecation,etc.With the increase of nebulization times of ovalbumin,the mental state of rats gradually deteriorated,the hair of rats was gradually fluffy,the glossiness of hair gradually decreased,and the rate of weight gain slowed down or weight loss.Rats in each treatment group showed better general state than those in the model group after taking drugs.(3)Detection of airway resistance of rats in each group:in the state of normal saline:compared with the blank group,the resistance of expiration in the model group increased(P<0.05).Compared with the model group,each administration group had a larger rat expiratory airway resistance decreased(P<0.05).Intravenous injection of acetylcholine(0.025mg/kg,0.05 mg/kg,0.2 mg/kg):compared with the blank control group,the resistance of expiration in the model group rats increased(P<0.05).Compared with the model group,the resistance of expiration of the rats in each administration group decreased(P<0.05).Intravenous injection of acetylcholine(0.1 mg/kg):compared with the blank control group,the resistance of expiration in the model group increased(P<0.05).Compared with the model group,the resistance of expiration of rats in the Chinese medicine high-dose group and the TRPV1 antagonist group decreased(P<0.05),and the resistance of expiration of rats in the other administration groups showed a decreasing trend(P>0.05).(4)ECP,IL-4,IL-5,TGF-?1 content of serum in rats:ECP:compared with the blank control group,the ECP level of serum in the model group was significantly increased(P<0.01).Compared with the model group,the ECP level of serum in each administration group was significantly reduced(P<0.01).IL-4:compared with the blank control group,the IL-4 level of serum in the model group was significantly increased(P<0.01).Compared with the model group,the IL-4 level of serum in the Chinese medicine middle-dose group and the TRPV1 antagonist group was reduced(P<0.05);IL-4 level of serum in the remaining administration groups tended to decrease(P>0.05).IL-5:compared with the blank control group,the IL-5 level of serum in the model group was significantly increased(P<0.01).Compared with the model group,the IL-5 level of serum in each administration group was significantly reduced(P<0.01).TGF-?1:compared with the blank control group,the TGF-31 level of serum in the model group was significantly increased(P<0.01).Compared with the model group,the TGF-31 level of serum in each administration group was significantly reduced(P<0.01).(5)ECP,IL-4,IL-5,TGF-31 content of bronchoalveolar lavage fluid in rats:ECP:compared with the blank control group,the ECP level of bronchoalveolar lavage fluid in the model group increased(P<0.05).Compared with the model group,the ECP level of bronchoalveolar lavage fluid in each administration group has a downward trend(P>0.05).IL-4:compared with the blank control group,the IL-4 level of the bronchoalveolar lavage fluid in the model group was significantly increased(P<0.01).Compared with the model group,the IL-4 level of the bronchoalveolar lavage fluid in each administration group was significantly reduced(P<0.01).IL-5:compared with the blank control group,the IL-5 level of bronchoalveolar lavage fluid in the model group was significantly increased(P<0.01).Compared with the model group,the IL-5 level of rats in the high-dose group of traditional Chinese medicine decreased(P<0.05),while the IL-5 level of rats in the other administration groups decreased significantly(P<0.01).TGF-?1:compared with the blank control group,the TGF-?1 level of bronchoalveolar lavage fluid in the model group was significantly increased(P<0.01).Compared with the model group,the TGF-?1 level in the high-dose and low-dose groups of traditional Chinese medicine rats was significant decreased(P<0.01),while the levels of TGF-?1 decreased in the glucocorticoid group,the Chinese medicine middle-dose group,and the TRPV1 antagonist group(P<0.05).(6)TRPV1 protein expression in lung tissue of rats in each group:compared with blank control group,TRPV1 protein expression in lung tissue of model group showed an increasing trend(P>0.05).Compared with model group,TRPV1 protein expression in rats of each administration group showed a decreasing trend in different degrees(P>0.05).Conclusion:(1)TRPV1 may be involved in the development of CVA.(2)Jin Fei Zhi Ke soup can reduce airway resistance and improve airway hyperresponsiveness in CVA rats.(3)Jin Fei Zhi Ke soup may reduce the content of ECP,IL-4,IL-5,and TGF-?1 by downregulating the expression of TRPV1 protein in the lung tissue of CVA rats,thereby reducing chronic airway inflammation.