Berberine Protects Against Simulated Ischemia/Reperfusion Injury-Induced H9C2 Cardiomyocytes Apoptosis In Vitro and Myocardial Ischemia/Reperfusion-Induced Apoptosis In Vivo by Regulating the Mitophagy-Mediated HIF-1α/BNIP3 Pathway.

Na Zhu,Peiyuan Hao,Yuwei Zhang,Yongli Li,Li Li,Qiubo Du,Xueming Cao,Jiang Li,Jinying Li

doi:10.3389/fphar.2020.00367

Abstract

Berberine (BBR) has a variety of pharmacological activities and is widely used in Asian countries. However, the clinical application of BBR still lacks scientific basis, what protective mechanism of BBR against myocardial ischemia-reperfusion injury (MIRI). In vitro experiments, BBR pretreatment regulated autophagy-related protein expression, induced cell proliferation and autophagosome formation, and reduced the mitochondrial membrane potential (ΔΨm) increase in H9C2 cells. In vivo experiments, BBR reduced the myocardial infarct size, decreased cardiomyocyte apoptosis, and markedly decreased myocardial enzyme (CK-MB, LDH, and AST) activity-induced I/R. In addition, upon BNIP3 knockdown, the regulatory effects of BBR on the above indicators were weakened both in H9C2 cells and in vivo. Luciferase reporter and ChIP assays indicated that BBR mediated BNIP3 expression by enhancing the binding of HIF-1α to the BNIP3 promoter. BBR protects against myocardial I/R injury by inducing cardiomyocytes proliferation, inhibiting cardiomyocytes apoptosis, and inducing the mitophagy-mediated HIF-1α/BNIP3 pathway. Thus, BBR may serve as a novel therapeutic drug for myocardial I/R injury.

Highlights

Coronary heart disease (CHD) has become the leading cause of human death, accounting for 13.2% of the top 10 causes (Lang et al, 2017)
Research has shown that the HIF-1a/BNIP3 pathway is important in hypoxia-induced autophagy in myocardial ischemia-reperfusion injury (MIRI) (Bruick, 2000; Tracy et al, 2007; Farrall and Whitelaw, 2009; Feng et al, 2016; Liu et al, 2019)
There were no significant changes in myocardial enzyme activity between the shBNIP3+BBR+I/R group and the I/ R group, indicating that BBR could not effectively regulate myocardial enzyme activity when BNIP3 was silenced. These results show that BBR reduces I/R-induced cardiomyocyte apoptosis and alleviates MIRI injury via regulating BNIP3-mediated mitochondrial autophagy

Summary

Introduction

Coronary heart disease (CHD) has become the leading cause of human death, accounting for 13.2% of the top 10 causes (Lang et al, 2017). The most effective therapeutic intervention for CHD is timely and effective myocardial reperfusion. The process of myocardial reperfusion can itself induce further cardiomyocyte death and result in myocardial ischemiareperfusion injury (MIRI) (Britto et al, 2018). There remains no effective clinical therapy for preventing myocardial reperfusion injury (Hausenloy and Yellon, 2013). MIRI research has shown that mitochondrial autophagy can eliminate damaged mitochondria and maintain intracellular environment homeostasis and cardiomyocyte function in MIRI and that mitochondrial autophagy regulation is an effective method for protecting against MIRI (Dong et al, 2010; Yang et al, 2010).

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