Effect Of Kangxian Yixin Decoction On Myocardial Damage Caused By Immunity Based On Th17/Treg Cell Balance

Purpose:Kangxian Yixin Decoction（KYD）is an effective prescription for clinical treatment of dilated cardiomyopathy（DCM）.The preliminary basic research found that this prescription can improve myocardial injury and inhibit ventricular remodeling,but its specific mechanism of action is still unclear.Recent studies have shown that autoimmunity can cause myocardial damage,and Th17/Treg cell imbalance has a certain role in myocardial immune damage.Therefore,this study intends to replicate the myocardial immune damage model caused by porcine myosin to observe the effect of KYD on myocardial immunity and explore the potential mechanism of action.Method:1.With reference to literature research methods,the model of myocardial immune injury was replicated by injecting porcine myocardial myosin into multiple places in the groin,underarms and back.After 6 weeks,the modeled m ice were randomly divided into a model group,a KYD group and a valsartan group,and a normal control group.The related indicators were tested after continuous gastric administration for 4 weeks.2.Observe the changes in the size of the heart and spleen in each group,and detect the pathological changes of myocardial tissue by HE and Masson staining.3.Enzyme-linked immune sorbent assay detects the expression of BNP,AMA,inflammatory factor IL-17 and IL-10 in mouse serum.4.Western Blotting method was used to detect the expression levels of myocardial tissue apoptosis indicators Bax and Bcl2 protein.5.Isolate mouse spleen lymphocytes,and detect the expression level of Th17and Treg cells in the spleen by flow cytometry.6.Detect the expression of RORγt,Foxp3 in the spleen by Real-time PCR.Result:1.After 6 weeks of modeling,the heart function of the mice in each group was detected by ultrasound.Compared with the normal group,the LVEDD and LVESD of the model group increased,and the EF decreased.The mice with significant changes in heart function were randomly divided into the model group（n=12）.KYD group（n=12）and valsartan group（n=12）.2.The results of myocardial histopathological staining showed that compared with the normal group of mice,the myocardial cells in the myocardial tissue of the model group were enlarged,degenerated and necrotic.There was obvious inflammatory cell infiltration in the myocardial interstitium,increased deposition of collagen,and myocardial fibers obviously.After the intervention of Kangxian Yixin Decoction,myocardial cell edema and degeneration and necrosis were reduced,myocardial interstitial inflammatory cell infiltration decreased,and myocardial tissue interstitial collagen fibers decreased.3.Enzyme-linked immune sorbent assay was used to detect the expression of BNP,anti-myocardial antibodies,IL-17,and IL-10.Compared with the normal group,IL-17 increased and IL-10 decreased in the model group,and the difference was statistically significant（P<0.05）.Compared with the model group,IL-17 decreased and IL-10 increased in KYD group,the difference was statistically significant（P<0.05）4.Western Blotting was used to detect apoptosis indicators Bax and Bcl2 in myocardial tissue.The results showed that compared with the normal group,the ratio of Bax/Bcl2 in the myocardial tissue of the model group increased（P<0.05）;while the myocardium of the mice treated with KYD The ratio of Bax/Bcl2 in tissues decreased（P<0.05）5.The results of flow cytometry detection of Th17/Treg cells in the spleen of mice showed that compared with the normal group,Th17 cells in the spleen of the model group increased,Treg cells decreased,and Th17/Treg cell ratio increased（P<0.05）,while KYD After the intervention,Th1 7 cells in the spleen of mice decreased,Treg cells increased,and the ratio of Th17/Treg cells decreased（P<0.05）.6.Real-time PCR detection of RORγt mRNA and Foxp3 mRNA in the spleen showed that compared with the normal group,the expression of RORγt mRNA in the spleen of the model group increased,the expression of Foxp3 mRNA decreased,and the ratio of RORγt/Foxp3 increased（P<0.05）.Compared with the model group,the expression of RORγt mRNA in the spleen of mice in the Kangxianyixin prescription group decreased,the expression of Foxp3 mRNA increased,and the ratio of RORγt/Foxp3 decreased（P<0.05）Conclusion:1.KYD can improve the pathological changes of myocardial tissue in mice with myocardial immune damage,down-regulate the ratio of myocardial cell apoptosis factor Bax/Bcl₂,reduce the expression of myocardial injury index BNP,and inhibit the expression of immune factor anti-myocardial antibodies,suggesting KYD may improve myocardial damage in mice by regulating immunity.2.The mechanism of KYD for improving myocardial immune damage may be related to its reduction of the ratio of Th17/Treg cells and RORγt/Foxp3 Mrna in the spleen of mice with myocardial immune damage,reducing the expressi on of IL-17and increasing the expression of IL-10.