Experimental Study On The Protective Effect Of Wushen Erlian Granule On Viral Myocarditis Myocardial Cell

Objective:To study the protective effect of Wushen Erlian granule on myocardial cells of neonatal rats and mice with viral myocarditis by in-vivo and in-vitro experiment.Methods:In the in-vitro experiment,Wistar myocardial cells of neonatal rats are prepared and divided randomly into two groups after culture.One group is used to test the toxicity of Wushen Erlian granule myocardial cells and the other group,after being artificially inoculated with Coxsackie virus B3(CVB3),is divided randomly into normal group and a virus group.Wushen Erlian granule group and ribavirin groups of different dose.ELISA is used to detect myocardial injury markers(troponin 1,myohemoglobin and creatine kinase isoenzyme).MTT is used to detect the antiviral effects of Wushen Erlian granule.In the in-vivo experiment,BalB/c mice model with viral myocarditis is built.Mice are divided randomly into a model,normal,ribavirin and Wushen Erlian granule groups of large,middle and small dose.Each group with 10 mice,are continuously administrated with drug by drenching for six days.To observe their general conditions during the whole process and cease the experiment two hours after the last administration,they are weighed.Then the specimen is collected according to testing requirements.Then use the collected specimen for myocardial enzyme indicator detection,viscera index detection,NK killer cells detection and the pathological detection of myocardial tissue.Result:Wushen Erlian granule and ribavirin median toxic concentration(TC50)are,respectively,415.28±1.82μg/mL,376.49 ±2.77μg/m.After Coxsackie virus B3 infects the myocardial cells of the neonatal rats.The index of cardiac injury under ELISA detection shows that compared with the model group and ribavirin group,the Wushen Erlian granule of 250μg/mL has a significantly reduced concentration(P<0.05)of creatine kinase-MB(CK-MB),cardiac troponin(cTnI)and myoglobin(MYO),indicating that Wushen Erlian granule of this concentration could significantly reduce myocardial cell injury caused by virus infection.Through MTT detection on antiviral effects of Wushen Erlian granule,the result shows that therapeutic index of Wushen Erlian granule on viral myocarditis is 13.02±0.94,and in-vitro antiviral therapeutic index of Wushen Erlian granule is significantly higher than that of the ribavirin group(6.93±0.93)(P<0.05).Through light microscopy detection in myocardial tissue,the result shows that mice in the model group have a large number of inflammatory cell infiltration under the myocardial ectoblast.Myocardial necrosis could be seen in the foci.Myocardial cell swelling and degeneration,and muscle fiber fracture could occasionally be seen.Compared with normal group,the pathological score in model group is 3.02±0.3,P<0.01,proving a successful model.And,the pathological score of large and mid-dose Wushen Erlian granule groups,compared with ribavirin group,P<0.01,indicating that Wushen Erlian granule could reduce the impairment of Coxsackie virus B3 in the myocardial tissue of mice.The spleen and thymus index in large and mid-dose Wushen Erlian granule groups and ribavirin group show signs of increase,indicating that the tested drug has significant effect in regulating immunization.The detection result of NK killing effect shows that both large and mid-dose Wushen Erlian granule could improve the killing capacity of NK cells and in comparison with the model group and the normal group,P<0.05.Through myocardial enzyme detection in the mice,the result shows that CK-MB and cTnl in large and mid-dose Wushen Erlian granule group,model group,and ribavirin group are significantly reduced(P<0.05),indicating that large and mid-dose Wushen Erlian granule could protect myocardial cells and reduce the impairment resulted from Coxsackie virus B3,thus beneficial for the myocardial repair.Conclusion:Wushen Erlian granule could inhibit the proliferation of Coxsackie virus B3 to some extent,reduce the impairment resulted from Coxsackie virus B3,stimulate the body’s immune regulation mechanism,and provide certain protective effects on myocardial cells.