Astragalus Root dry extract restores connexin43 expression by targeting miR-1 in viral myocarditis

Abstract

BackgroundViral myocarditis is defined as viral infection of myocardial tissue leading to impaired heart function and heart failure. Accumulating evidences have shown that arrhythmia is one of important complicating diseases of viral myocarditis causing increased mortality and morbidity. There are no effective treatment for the viral infection and complicating arrhythmia. PurposeThis study investigated the effect and mechanism of Astragalus Root dry extract (ARDE) on arrhythmia induced by CVB3 in mice. MethodsThe mice and HL-1 cells were treated with CVB3 and ARDE. Reciprocal regulation of Cx43 and miR-1 were observed in the CVB3 infected mouse myocardium and culture HL-1 cells. ResultsCVB3 IP injection increased immune cell infiltration in mouse left ventricle and caused irregular arrhythmia. ARDE treatment prevented the increase of immune cell infiltration and arrhythmia. Overexpression of miR-1 significantly inhibited both endogenous Cx43 expression and Cx43 3′UTR luciferase activity in HL-1 cells. Mutation of census binding site of +1586–1593 bp not +465–472 bp in Cx43 3′UTR luciferase resulted in abolishment of miR-1 inhibitory effects in HL-1 cells. Loss-of- function of miR-1 restored CVB3-induced Cx43 expression reduction in cultured HL-1 cells. The presence of ARDE attenuated the augmented miR-1 induced by CVB3 infection in vivo and in vitro. ConclusionThis study identified that CVB3 infection reduced Cx43 expression by elevating miR-1 level in mouse viral myocarditis. For the first time, ARDE was shown to prevent arrhythmia, and rescue CVB3-induced endogenous Cx43 expression by regulating miR-1 level.