Ghrelin通过诱导细胞自噬加剧H9c2细胞缺氧-复氧损伤

Detrimental effect of Ghrelin on hypoxia/reoxygenation- induced injury of H9c2 cells by enhancing cell autophagy

用CoCl_2诱导H9c2细胞缺氧损伤24 h后,再将培养基换成新鲜培养基,以模拟心肌细胞体外缺血-再灌模型,并在此基础上通过ghrelin对细胞自噬的调节来评价ghrelin对心肌细胞缺氧-复氧损伤的调节作用.用流式细胞技术和LDH活性检测细胞凋亡和坏死.WST-1检测细胞活力;DCFH2-DA探针评估细胞内活性氧水平;用Western blotting检测细胞自噬.研究结果表明ghrelin处理的缺氧-复氧损伤细胞活力降低、LDH活性、细胞凋亡、ROS含量及自噬增加,用3MA处理后可明显抑制细胞自噬,并伴随细胞活力的显著升高,因此,ghrelin通过加强细胞自噬加剧了CoCl_2诱导的H9c2细胞缺氧-复氧损伤.

To evaluate the effect of ghrelin on myocardial ceils by regulating autopahgy, H9c2 cells were treated by COC12 for 24 h to induce hypoxic injury, then replaced with fresh medium to mimic I/R condi- tion in cardiac H9c2 ceils in vitro. Cell apoptosis and necrosis were evaluated by the Flow cytometry as- say and LDH activity. Cell viability was detected by WST-1 assay; ROS levels were assessed using DCFH2-DA; and LC3II was measured by Western blot analysis. The results from above assays showed that ghrelin treatment significantly intensifies Hypoxia/Reoxgenation （H/R） injury by increasing H9c2 cell apoptosis, LDH activity and ROS content and cell autophagy levels. Further experiments revealed that inhibiting autophagy by 3MA almost abrogated the induction of autophagy by ghrelin, associated with a significant increase in cell viability. The results indicate that ghrelin intensifies H/R injured car- diac myocytes by enhancing autophagy.