Ischemia-reperfusion Injury Up-regulates Pim-3 Gene Expression in Myocardial Tissue
|School||Huazhong University of Science and Technology|
|Course||Cardiovascular within science|
|Keywords||Pim-3 Reperfusion injury H2O2 Apoptosis|
Objective : To observe the ischemia-reperfusion injury in rat myocardial tissue Pim-3 gene expression , and to explore the possible molecular mechanisms . Method: 1 ) extraction of SD rat tissues , using RT-PCR, Weston blot detection of Pim-3 gene expression in tissue distribution , immunohistochemical detection of Pim-3 gene expression in cardiac tissue . 2 ) Take 30 SD male adult rats were randomly divided into five groups , sham, ligation of the left anterior descending (LAD) coronary artery 15min, 30min, 30min ligation and reperfusion 30min, 30min ligation and reperfusion 120min, left ventricular tissue specimens , using by RT-PCR and immunohistochemistry assay Pim-3 gene expression changes . 3 ) primary cultured neonatal rat myocardial cells and with different concentrations of H2O2 (0umol/ml, 5umol/ml, 10umol/ml, 20umol/ml), tumor necrosis factor -α (0ng/ml, 1ng/ml, 5ng/ml , 10ng/ml) treatment, using RT-PCR method to detect the myocardial cells Pim-3 mRNA and protein expression changes . 4 ) transfected Pim-3siRNA interference fragment into neonatal rat cardiomyocytes , and use H2O2-induced apoptosis , detected changes in myocardial apoptosis . Results: 1 ) normal myocardial tissue expression of Pim-3 mRNA and protein ; 2 ) ischemia and reperfusion injury were caused by myocardial tissue Pim-3 gene expression was elevated ; 3) H2O2 increased primary cultured cardiomyocytes Pim-3 gene expression, whereas TNF-α does not affect cardiac myocytes Pim-3 gene expression ; 4) Pim-3 gene silencing can promote cardiomyocyte apoptosis induced by H2O2 . Conclusion: ischemia-reperfusion injury through oxidative stress signaling pathways increase myocardial tissue Pim-3 gene expression .