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Proanthocyanidin protects H9C2 cells against hypoxia/reoxygenation injury via JAK2/STAT3 signaling pathway

YU Chen-Bin, ZHAO Guo-Long, YU Li-Ming, YU Shi-Qiang, DUAN Wei-Xun, ZHANG Hai-Feng

Emergency Department of Jiangsu Research Institute of Traditional Chinese Medicine, Jiangsu 210028； Experimental Teaching Center, The Fourth Military Medical University, Xi’an 710032, China； Department of Cardiovascular Surgery, Xijing Hospital, Fourth Military Medical University, Xi’an 710032, China

Abstract

The present study was aimed to investigate the underlying mechanisms of the protective effect of proanthocyanidin (Pro) against hypoxia/reoxygenation (H/R) injury in H9C2 cells with a focus on Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3) signaling pathway. H9C2 cells were randomly assigned to 5 groups, including the control group (Con), the H/R-injured group (H/R), the Pro-treated group (H/R+Pro), the JAK2 siRNA-treated group (H/R+Pro+JAK2 siRNA) and the JAK2 siRNA control group (H/R+JAK2 siRNA). The cells were pretreated with Pro (40 µmol/L) for 8 h before 2 h of hypoxia and 4 h of reoxygenation. Cellular viability and apoptosis rate were detected by MTT and TUNEL methods, and superoxide generation was measured. JAK2/STAT3 signaling, oxidative stress markers and endoplasmic reticulum stress markers were also detected by Western blot. We found that Pro treatment significantly improved cellular viability and reduced apoptosis rate in H/R-treated H9C2 cells. In addition, Pro treatment significantly up-regulated the phosphorylation levels of JAK2 and STAT3, down-regulated the superoxide generation, gp91phox, glucose-regulated protein 78 (GRP78), CCAAT/enhancer binding protein homologous protein (CHOP) and caspase-12 expression. However, these protective effects of Pro were all attenuated by JAK2 siRNA administration. Taken together, we demonstrated that Pro protects H9C2 cells against H/R-induced oxidative stress and endoplasmic reticulum stress injury via JAK2/STAT3 signaling pathway.

Key words: proanthocyanidin; hypoxia/reoxygenation injury; JAK2/STAT3; oxidative stress; endoplasmic reticulum stress

Received: 2016-02-25　　Accepted: 2016-04-28

Corresponding author: 段维勋,张海锋　　E-mail: duanweixun@126.com,hfzhang@fmmu.edu.cn

Citing This Article：

YU Chen-Bin, ZHAO Guo-Long, YU Li-Ming, YU Shi-Qiang, DUAN Wei-Xun, ZHANG Hai-Feng. Proanthocyanidin protects H9C2 cells against hypoxia/reoxygenation injury via JAK2/STAT3 signaling pathway. Acta Physiol Sin 2016; 68 (5): 568-574 (in Chinese with English abstract).