ZFP580, a novel transcription factor,is involved in cardioprotection of hypoxic preconditioning against hypoxia-reoxygenation injury in myocardial cells

AIM：To elucidate whether ZFP580 is involved in the cardioprotective effects of hypoxic preconditioning (HPC) against hypoxia-reoxygenation (H/R) injury in H9c2 myocardial cells. METHODS：Rat heart-derived H9c2 cells were cultured in DMEM. H/R was induced by incubation under ischemic hypoxia for 3 h and reoxygenation for 2 h. HPC was induced by exposing the H9c2 cells to 10 min of hypoxia and 20 min of reoxygenation for 3 cycles before H/R treatment. MTT staining and LDH leakage detection were used to evaluate the effects of HPC. Western blotting was used to detect the protein levels of ZFP580, phosphorylated ERK1/2 and cleaved caspased-3. The effects of ZFP580 overexpre-ssion or knockdown on H/R induced apoptosis were determined. RESULTS：The results of MTT staining and LDH leakage detection showed evidence of HPC cytoprotection against H/R-induced cell death in H9c2 cells. ZFP580 protein level and ERK1/2 phosphorylation were significantly increased in the HPC group compared with control group and H/R group. PD98059, an inhibitor of ERK1/2 phosphorylation, significantly suppressed the HPC-induced up-regulation of ZFP580 protein expression. ZFP580 overexpression significantly inhibited apoptosis and caspase-3 activation in H9c2 cells. CONCLUSION：HPC exhibits cytoprotection against H/R and leads to high level of ZFP580 protein in H9c2 cells. ZFP580 is regulated by ERK1/2 activation and mediates the anti-apoptotic effect of the ERK1/2 signaling pathway in HPC cytoprotection.