Li Qi1, Qi Xin2, Jia Wenjun1. 1. Department of Cardiology, Tianjin Union Medical Center, Tianjin 300121, China. 2. Department of Cardiology, Tianjin Union Medical Center, Tianjin 300121, China. Electronic address: qixinstudy@163.com.
Abstract
BACKGROUND: Coxsackievirus B3 (CVB3), a member of the picornavirus family, is one of the major causative enteroviruses of viral myocarditis. The aim of the current study was to investigate the role and underlying mechanism of iNOS and autophagy in CVB3 infected cardiomyocytes. METHODS: Myocardial cell H9c2 were randomly divided into four groups: control group, CVB3 group, CVB3+L-NAME group and the CVB3+iNOS siRNA group. Cell proliferation was detected by MTT method and cell apoptosis was determined by flow cytometric. The protein expression levels were determined by Western blot. Anisomycin was used to activate JNK pathway in CVB3 infected H9c2 cells. RESULTS: The results demonstrated that the inhibition of iNOS significantly elevated cell proliferation and suppressed cell apoptosis of CVB3-induced H9c2 cells. The production of MDA was obviously decreased, while the activity of SOD was increased by the addition of L-NAME or iNOS siRNA compared with the CVB3 group. Expression of the autophagy marker proteins LC3 II and Beclin 1 was significantly decreased, and the autophagy substrate p62 was dramatically increased in iNOS inhibition groups compared with the CVB3 group. Moreover, iNOS inhibition suppressed the JNK pathway in CVB3-infected H9c2 cells. Furthermore, administration of the JNK pathway stimulator, anisomycin, counteracted the effect of iNOS inhibition in CVB3-infected H9c2 cells. CONCLUSION: The inhibition of iNOS protects cardiomyocytes against CVB3-induced cell injury by regulating autophagy and the JNK pathway, which may provide a novel therapeutic strategy for treating CVB3-induced myocarditis.
BACKGROUND:Coxsackievirus B3 (CVB3), a member of the picornavirus family, is one of the major causative enteroviruses of viral myocarditis. The aim of the current study was to investigate the role and underlying mechanism of iNOS and autophagy in CVB3 infected cardiomyocytes. METHODS: Myocardial cell H9c2 were randomly divided into four groups: control group, CVB3 group, CVB3+L-NAME group and the CVB3+iNOS siRNA group. Cell proliferation was detected by MTT method and cell apoptosis was determined by flow cytometric. The protein expression levels were determined by Western blot. Anisomycin was used to activate JNK pathway in CVB3 infected H9c2 cells. RESULTS: The results demonstrated that the inhibition of iNOS significantly elevated cell proliferation and suppressed cell apoptosis of CVB3-induced H9c2 cells. The production of MDA was obviously decreased, while the activity of SOD was increased by the addition of L-NAME or iNOS siRNA compared with the CVB3 group. Expression of the autophagy marker proteins LC3 II and Beclin 1 was significantly decreased, and the autophagy substrate p62 was dramatically increased in iNOS inhibition groups compared with the CVB3 group. Moreover, iNOS inhibition suppressed the JNK pathway in CVB3-infected H9c2 cells. Furthermore, administration of the JNK pathway stimulator, anisomycin, counteracted the effect of iNOS inhibition in CVB3-infected H9c2 cells. CONCLUSION: The inhibition of iNOS protects cardiomyocytes against CVB3-induced cell injury by regulating autophagy and the JNK pathway, which may provide a novel therapeutic strategy for treating CVB3-induced myocarditis.
Authors: Michael F Bode; Clare M Schmedes; Grant J Egnatz; Vanthana Bharathi; Yohei M Hisada; David Martinez; Tomohiro Kawano; Alice Weithauser; Leah Rosenfeldt; Ursula Rauch; Joseph S Palumbo; Silvio Antoniak; Nigel Mackman Journal: Sci Rep Date: 2021-07-12 Impact factor: 4.996