| Objectives:Objective to analyze the clinical data of EOS-AECOPD and explore its TCM syndrome characteristics,so as to provide objective basis for TCM syndrome differentiation of EOS-AECOPD.To evaluate the clinical effect of Houpu Mahuang Decoction in the treatment of EOS-AECOPD.To investigate the effect and mechanism of Houpu Mahuang Decoction on the model of EOS increased airway injury induced by ovalbumin(OVA)and cigarette smoke(CS).Methods:1.By sorting out the understanding of the pathological products of phlegm,Yin,blood stasis and the pathogenesis of AECOPD,combing the etiology and pathogenesis of AECOPD,treatment,prescription and pathological product factors and the relationship between AECOPD,and combining with modern research to analyze the research status of EOS-AECOPD.2.To collect the clinical data of patients with EOS-AECOPD(peripheral blood eosinophil B-EOS count ?300cells/?L)and collect the four diagnostic information of traditional Chinese medicine,to study the clinical characteristics of EOS-AECOPD,and to study the regularity of syndrome and distribution characteristics of EOS-AECOPD by frequency analysis and cluster analysis.3.On the basis of the research on the regularity of syndromes and treatment methods and prescriptions of EOS-AECOPD,this paper took Houpu Mahuang Decoction as an example to explore its characteristics of syndrome and treatment,and used the methods of two-way cohort study to evaluate the clinical efficacy of Houpu Mahuang Decoction in the treatment of EOS-AECOPD.4.Objective to explore the effects and mechanism of Houpu Mahuang Decoction.(1)The eosinophilic airway injury model was established by OVA+CS.The inflammatory cell counts,lung function and lung pathological changes in peripheral blood and bronchoalveolar lavage fluid(BALF)of each group were compared.The gene and protein expression levels of tight junctions(TJs)(ZO-1,Occludin and Claudin-2)and TRPA1 in lung tissue were detected by qPCR and Western blot,respectively.OVA+CS was used to intervene TRPA1 knockout mice(TRPA1-/-)and compared with wild-type mice(WT).(2)Houpu Mahuang Decoction and its extracts were used to intervene OVA+CS mice model.The expression of inflammatory cells,lung function,lung tissue pathology,TJs,TRPA1 gene and protein in lung tissue of each group were observed,and the effect and mechanism of Houpu Mahuang Decoction on OVA+CS induced EOS increased airway injury model mice were explored.Results:1.Through combing the ancient and modern literature,it is recognized that the etiology and pathogenesis of AECOPD mainly involve exogenous factors,internal injury factors and pathological product factors.Internal injury factors include loss of healthy qi,improper diet and emotional disorder.Deficiency of healthy qi is the fundamental cause of AECOPD.Exogenous factors,improper diet and emotional disorder are the main inducing factors.The generation and induction of pathological factors such as phlegm,yin and blood stasis are the important causes of AECOPD recurrence and difficult to control.Therefore,it is particularly important to study the treatment and mechanism of pathological factors such as phlegm,yin and blood stasis.The increase of EOS level in AECOPD patients is closely related to the risk of acute exacerbation,and the etiology and pathogenesis may be related to pathological product factors.2.According to the clinical data of 163 cases of high B-EOS group and non-high B-EOS group,the results showed that male patients in both groups were more than 80%.The smoking index of high B-EOS group was lower than that of non-high B-EOS group,and the number of acute exacerbations per year was higher than that of non-high B-EOS group(P<0.01).There was no significant difference in gender,age,BMI,allergic history,smoking history,course of disease and other general information(P>0.05).The percentage of eosinophils and neutrophils in high B-EOS group were significantly higher than those in non high B-EOS group(P<0.05),but the number of neutrophils and the total number of leukocytes had no significant difference(P>0.05).The levels of CRP,PCT and IgE in the two groups were tested by chi square test according to the grade data.The results showed that there were significant differences in the grade distribution of PCT and IgE data between the two groups(P<0.05,P<0.01).The proportion of IgE level higher than the normal range in the high B-EOS group was significantly increased(P<0.01).The level of FEV1 in high B-EOS group was lower than that in non high B-EOS group(P<0.05).There was no significant difference in other indexes such as mMRC,CAT,GOLD grade and pO2,pCO2(P>0.05).The clinical data of 43 cases of high B-EOS group and 61 cases of non high B-EOS group were analyzed statistically.The highest frequency of syndromes in high B-EOS group were gasp,cough,phlegm and white,tongue coating,yellow and white constipation,chest tightness,pulse floating,tongue coating,fatigue,fatigue,foam sputum,phlegm thinning,constipation,etc.Combined with syndrome differentiation,phlegm,Yin,dampness,heat and cold were the most common syndrome elements in high B-EOS group.Cluster analysis showed that lung was closely related to cold and Yin,followed by exterior and heat.3.Taking Houpu Mahuang Decoction as the exposure factor,the patients with EOS-AECOPD were divided into exposure group(n=20)and non exposure group(n=20).A retrospective cohort study was conducted.The results showed that the main syndrome scores of gasp,cough,chest tightness,sputum volume and sputum color,and the secondary syndrome scores of fatigue,diet,sleep,fever and defecation in the exposure group were significantly improved compared with those before treatment.Compared with the non exposure group,the quality,diet and stool of the patients in the exposure group improved more significantly,and the total score of syndrome decreased more significantly(P<0.05,P<0.01).The weight of syndrome elements of the two groups were compared.The results showed that the two groups could significantly reduce the lung,surface,cold,drink,phlegm and heat syndrome elements(P<0.05,P<0.01),but the Qi deficiency and dampness syndrome elements were not significantly reduced,but the dampness syndrome elements of the non exposure group were increased.After treatment,cold,Yin,Qi deficiency and dampness syndrome elements in the exposed group decreased more significantly than those in the non exposed group(P<0.05).4.The experimental results show that compared with NS group,the levels of EOS in peripheral blood,EOS and NEU in BALF were significantly increased in OVA+CS group(P<0.01),and the pulmonary function ventilation(MV),inspiratory peak flow(PIF)and expiratory peak flow(PEF)were significantly decreased in OVA+CS group.The expressions of ZO-1 and Occludin were significantly down regulated(P<0.01),and the expression of Claudin-2 was up-regulated(P<0.01),while the levels of TRPA1 gene and protein were significantly up-regulated(P<0.01).In addition,compared with WT mice,the inflammatory indexes,lung injury and TJs damage were reduced or eliminated after TRPA1 gene knockout(P<0.05,P<0.01).After administration,Houpu Mahuang Decoction,especially the high-dose and ethyl acetate group,could significantly improve the symptoms of model mice.The levels of EOS and other inflammatory cells in peripheral blood and BALF of Houpu Mahuang Decoction group were significantly lower than those of model group(P<0.01),and the indexes of lung function were also higher than those of model group(P<0.05,P<0.01)The expression of TRPA1 gene and protein in lung tissue of Houpu Mahuang Decoction group were significantly decreased(P<0.05,P<0.01),and the expression level of TJs protein were also improved(P<0.05,P<0.01).Conclusion:1.Through combing the ancient and modern literature,this study realized that phlegm,yin and blood stasis are the most common pathological products in the process of COPD,and they are also the main pathological factors leading to AECOPD.To some extent,they determine the severity of the disease and the prognosis.The increase of EOS level in AECOPD patients may be related to the pathological product factors.2.The etiology and pathogenesis of EOS-AECOPD patients were cold drinks and volts.After being induced by exogenous pathogenic factors or internal causes,drinking and depression become hot,and drinking water was spread to the lungs.Symptoms could be seen as gasp,cough,phlegm and white,tongue coating,yellow and white constipation,chest tightness,pulse floating,tongue coating,fatigue,fatigue,foam sputum,phlegm thinning,constipation,etc.According to the analysis of common clinical prescriptions of cold drink for lung retention and Yin depression for heat syndrome,taking Houpu Mahuang Decoction as an example,the clinical observation of Houpu Mahuang Decoction in the treatment of EOS-AECOPD patients were carried out by using a two-way cohort study.The results showed that Houpu Mahuang Decoction exposure group could significantly improve the syndrome score and syndrome element weight of EOS-AECOPD.It not only had a significant advantage in Xuanfei Huayin and Xiaqi Jiangni,but also could achieve the purpose of invigorating spleen and Qi by eliminating phlegm,dampness and other pathological factors.It could shorten the hospitalization time of patients with EOS-AECOPD,and provided a certain clinical basis for the selection of Houpu Mahuang Decoction in the treatment of patients with EOS-AECOPD.3.Houpu Mahuang Decoction could reduce the inflammatory indexes and lung injury of EOS increased model mice induced by OVA combined with CS,and the expression of ZO-1,Occludin,Claudin-2 were also re-regulated.The expression of TRPA1 in Houpu Mahuang Decoction group was decreased,and the effect of high dose and ethyl acetate extract group were the most significant.At the same time,the injury of TJs protein(ZO-1,Occludin,Claudin-2)and airway inflammation and injury were not significant in TRPA1-/-group.These results suggest that Houpu Mahuang Decoction can protect lung tissue by reducing the damage of epithelial barrier and airway inflammation,which may be related to the regulation of TRPA1 channel. |
PDF Full Text Request