HDAC3 通过靶向 GSK-3β 介导瑞芬太尼后处理对缺氧/复氧损伤 H9c2 心肌细胞的心脏保护作用。

HDAC3 Mediates Cardioprotection of Remifentanil Postconditioning by Targeting GSK-3β in H9c2 Cardiomyocytes in Hypoxia/Reoxygenation Injury.

机构信息

Department of Anesthesiology, The first Affiliated Hospital of Anhui Medical University, Hefei, China.

Department of Anesthesiology, The First Affiliated Hospital, Medical School of Zhejiang University, Hangzhou, China.

出版信息

Shock. 2018 Aug;50(2):240-247. doi: 10.1097/SHK.0000000000001008.

DOI:10.1097/SHK.0000000000001008

PMID:28957873

Abstract

BACKGROUND

Remifentanil postconditioning (RPC) confers robust cardioprotection against ischemia/reperfusion (I/R) injury. We recently determined that HDAC3 was involved in RPC-induced cardioprotection. However, the role of HDAC3 and its possible mechanisms in RPC-induced cardioprotection are unknown, which we aimed to evaluate in an in vitro hypoxia/reoxygenation (HR) model.

METHODS

Myocardium I/R injury was established after HR with H9c2 cardiomyoblasts. Cell viability and apoptosis were evaluated usingCCK-8 and flow cytometry of HR-injured cardiomyoblasts treated with or without RPC. Furthermore, effects of RPC on HDAC3 protein and mRNA expression were evaluated with Western blot and quantitative real-time PCR analyses, whereas GSK-3β expression was measured with Western blot.

RESULTS

RPC increased cell viability and reduced cell apoptosis (P < 0.05) in H9c2 cardiomyoblasts subjected to HR injury. In addition, RPC promoted the phosphorylation of GSK-3β at Ser9 site (P < 0.05) and suppressed the protein and mRNA expression of HDAC3 (P < 0.05). Lentiviral-transduced overexpression of HDAC3 had no significant effects on HR injury while attenuating the cardioprotective effects of RPC on cell viability and apoptosis (P < 0.05), GSK-3β phosphorylation (P < 0.05) in H9c2 cardiomyoblasts.

CONCLUSIONS

RPC attenuates apoptosis in H9c2 cardiomyoblasts after HR injury by downregulating HDAC3-mediated phosphorylation of GSK-3β. Our findings suggest that HDAC3, and its cross talk function with GSK-3β, may be a promising target for myocardium I/R injury.

摘要

背景

瑞芬太尼后处理（RPC）对缺血/再灌注（I/R）损伤具有强大的心脏保护作用。我们最近发现 HDAC3 参与了 RPC 诱导的心脏保护。然而，HDAC3 及其在 RPC 诱导的心脏保护中的可能机制尚不清楚，我们旨在体外缺氧/复氧（HR）模型中评估这些机制。

方法

用 H9c2 心肌细胞建立 HR 后的心肌 I/R 损伤模型。用 CCK-8 和 HR 损伤的心肌细胞的流式细胞术评估细胞活力和凋亡。此外，用 Western blot 和实时定量 PCR 分析评估 RPC 对 HDAC3 蛋白和 mRNA 表达的影响，用 Western blot 测定 GSK-3β的表达。

结果

RPC 增加了 HR 损伤的 H9c2 心肌细胞的活力，减少了细胞凋亡（P<0.05）。此外，RPC 促进了 GSK-3β 在 Ser9 位点的磷酸化（P<0.05），并抑制了 HDAC3 的蛋白和 mRNA 表达（P<0.05）。HDAC3 的慢病毒转导过表达对 HR 损伤没有显著影响，而减弱了 RPC 对细胞活力和凋亡的保护作用（P<0.05），以及 GSK-3β 的磷酸化（P<0.05）。