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目的探讨缺血后适应与心肌营养素-1(CT-1)对心肌细胞缺氧复氧的保护作用及其机制。方法将体外培养的H9C2细胞株分为正常对照组(a组)、缺氧复氧组(b组)、缺氧复氧+缺氧后适应组(c组)、缺氧复氧+缺氧后适应+CT-1组(d组)、缺氧复氧+缺氧后适应+CT-1+ERK抑制剂组(e组)、缺氧复氧+缺氧后适应+CT-1+二甲基亚砜组(f组),MTS法检测各组细胞存活率、流式细胞仪检测细胞凋亡率、Western blot检测细胞外调节激酶(ERK1/2)和磷酸化ERK1/2(p-ERK1/2)的表达、Q-PCR检测Bad mRNA的表达。结果与a组比较,其他各组细胞存活率下降,细胞凋亡率、Bad mRNA和p-ERK1/2(除外e组)表达量增加,P均<0.05;与b组比较,c组细胞存活率、p-ERK1/2表达量均增加,Bad mRNA表达、凋亡率下降,P均<0.05;与c组比较,d组细胞存活率、pERK1/2表达量均增加,细胞Bad mRNA表达、凋亡率下降(P均<0.05);与d组比较,e组细胞存活率下降,细胞凋亡率、Bad mRNA表达量增加,p-ERK1/2表达量下降;d、e组各项指标指标比较差异无统计学意义。结论缺氧后适应可减少心肌细胞缺氧复氧损伤,联合CT-1后保护作用明显加强,而ERK1/2抑制剂可阻断这种保护作用;该保护机制可能与CT-1激活ERK1/2信号通路,下调Bad mRNA的表达有关。
Objective To investigate the protective effects and mechanisms of ischemic postconditioning and cardiotrophin-1 (CT-1) on cardiomyocyte hypoxia and reoxygenation. Methods H9C2 cells were divided into normal control group (group a), hypoxia-reoxygenation group (group b), hypoxia-reoxygenation plus hypoxia adaptation group (group c), hypoxia- Hypoxia and reoxygenation + Hypoxia and reoxygenation + CT-1 + ERK inhibitor group (group e), Hypoxia and reoxygenation + Hypoxia and reoxygenation + CT-1 + Methyl sulfoxide group (group f). The cell viability was detected by MTS method and the apoptosis rate was detected by flow cytometry. The expressions of ERK1 / 2 and p-ERK1 / 2 were detected by Western blot. ERK1 / 2) expression, Bad mRNA expression was detected by Q-PCR. Results Compared with group a, the survival rates of other groups were decreased, the rates of apoptosis, Bad mRNA and p-ERK1 / 2 (except e group) increased, P <0.05; The expression of Bad-ERK1 / 2, the expression of Bad mRNA and the apoptosis rate of P-ERK1 / 2 increased significantly, P <0.05; (P <0.05). Compared with group d, the survival rate of group e decreased, the rate of apoptosis increased, the expression of Bad mRNA increased and the expression of p-ERK1 / 2 decreased. The indexes of group d and e No significant difference in indicators. CONCLUSION: Hypoxia can reduce the hypoxia-reoxygenation injury of cardiomyocytes. The protective effect of CTK-1 is obviously enhanced by ERK1 / 2 inhibitor. ERK1 / 2 inhibitor can block this protective effect. This protective mechanism may be related to the activation of ERK1 / 2 signaling pathway, down-regulated Bad mRNA expression.