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目的:探讨血红素加氧酶-1(HO-1)对H9c2心肌细胞氧化应激损伤的保护作用及其作用机制。方法:建立过氧化氢(H2O2)诱导的H9c2心肌细胞氧化应激损伤模型,给予HO-1诱导剂氯化血红素(Hemin)预处理,或给予HO-1抑制剂锌原卟啉(Znpp-IX)共孵育。双波长法检测HO-1活性;四甲基偶氮唑盐(MTT)法检测细胞存活率;Hoechst33342染色和Caspase-3活性检测评估细胞凋亡;硫代巴比妥酸显色法检测丙二醛(MDA)含量,氮蓝四唑显色法检测总超氧化物歧化酶(SOD)活性;Western Blot法检测NF-κB蛋白表达水平。结果:与H2O2组相比,Hemin显著增加细胞HO-1活性,且该作用能被Znpp-IX阻断。与H2O2组相比,HO-1活性增加能够显著下调细胞凋亡率和Caspase-3活性,降低细胞MDA含量,增加总SOD活性,且上述作用能被Znpp-IX逆转。此外,HO-1活性增加能够显著抑制H2O2诱导的NF-κB激活,且该作用能被Znpp-IX逆转。结论:Hemin诱导的HO-1活性增加可能通过抑制NF-κB激活,维持细胞氧化还原平衡状态,抑制H2O2诱导的H9c2心肌细胞氧化应激损伤。
Objective: To investigate the protective effect and mechanism of heme oxygenase-1 (HO-1) on oxidative stress injury of H9c2 cardiomyocytes. Methods: H2O2-induced oxidative stress injury model of H9c2 cardiomyocytes was established. Hemin preconditioning with HO-1 inducing agent or Znpp- IX) co-incubation. Dual-wavelength method was used to detect the activity of HO-1. Cell viability was detected by MTT assay. Cell apoptosis was assessed by Hoechst33342 staining and Caspase-3 activity assay. Thiobarbituric acid The content of MDA and the content of superoxide dismutase (SOD) were detected by the method of nitrogen blue tetrazolium colorimetry. The protein expression of NF-κB was detected by Western Blot. Results: Compared with H2O2 group, Hemin significantly increased the activity of HO-1, and this effect was blocked by Znpp-IX. Compared with H2O2 group, the increase of HO-1 activity could significantly reduce the apoptosis rate and Caspase-3 activity, decrease the MDA content and increase the total SOD activity, and the above effects could be reversed by Znpp-IX. In addition, increased HO-1 activity significantly inhibited H2O2-induced NF-κB activation, and this effect could be reversed by Znpp-IX. Conclusion: The increase of HO-1 activity induced by Hemin may inhibit the oxidative stress injury of H9c2 cardiomyocytes induced by H2O2 by inhibiting the activation of NF-κB, maintaining the redox balance of cells.