HMGB1 induces apoptosis and EMT in association with increased autophagy following H/R injury in cardiomyocytes

Ouyang,Fan; Huang,He; Zhang,Mingyu; Chen,Mingxian; Huang,Haobo; Huang,Fang; Zhou,Shenghua

doi:10.3892/ijmm.2016.2474

HMGB1 induces apoptosis and EMT in association with increased autophagy following H/R injury in cardiomyocytes

Authors:
- Fan Ouyang
- He Huang
- Mingyu Zhang
- Mingxian Chen
- Haobo Huang
- Fang Huang
- Shenghua Zhou
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Affiliations: Department of Cardiology, Xiangtan Central Hospital, Xiangtan, Hunan 411100, P.R. China, Department of Cardiology, The Second Xiangya Hospital of Central South University, Changsha, Hunan 410011, P.R. China
Published online on: January 29, 2016 https://doi.org/10.3892/ijmm.2016.2474
Pages: 679-689
Copyright: © Ouyang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Hypoxia/reoxygenation (H/R) is a critical factor in the pathogenesis of tissue injury following myocardial infarction (MI) which can lead to tissue damage and pathological remodeling. Therefore, it is necessary to try and prevent myocardial H/R injury in order to optimize the treatment of MI. This study aimed to explore the functions and molecular mechanisms of action of high mobility group box 1 (HMGB1) and its role in H/R injury to H9c2 cells. The mRNA expression of levels genes were detected by RT-qPCR. The protein levels were examined by western blot analysis. The Beclin 1 expression level was further determined by immunocytochemistry (ICC). In addition, an HMGB1 overexpression vector and a shRNA lentiviral vector were constructed in order to induce the overexpression and silencing of HMGB1, respectively. The apoptotic rate of the H9c2 cells was determined by flow cytometry. The expression of miR-210 was markedly increased following the exposure of the cells to H/R, thus indicating that the cell model of H/R injury was successfully established. In addition, an in vivo model of MI was also created using rats. The mRNA and protein level of HMGB1 was found to be upregulated in the myocardial tissue of the rats with MI and in the H9c2 cells subjected to H/R injury. HMGB1 promoted apoptosis by increasing the expression of cleaved caspase-3 and the apoptotic rate of the cells, while decreasing the expression of Bcl-2 during H/R in the H9c2 cells. HMGB1 promoted epithelial-to-mesenchymal transition (EMT) by reducing the protein level of the epithelial marker, E-cadherin, while increasing the expression of the mesenchymal markers, vimentin and fibroblast-specific protein (FSP), during H/R in the H9c2 cells. HMGB1 induced the apoptosis of the H9c2 cells and EMT following H/R in association with the induction of autophagy. HMGB1 induced autophagy by upregulating the expression of discoidin domain receptor 1 (DDR1) and downregulating the phosphorylation levels of mammalian target of rapamycin (mTOR). In conclusion, the findings of our study suggest that HMGB1 promotes apoptosis and EMT in association with the induction of autophagy through the upregulation of the expression of DDR1 and the downregulation of the phosphorylation of mTOR following H/R injury in H9c2 cells.

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