临床内科杂志 ›› 2019, Vol. 36 ›› Issue (11): 773-776.doi: 10.3969/j.issn.1001-9057.2019.11.018

• 临床基础研究 • 上一篇    下一篇

RNA干扰程序性细胞死亡因子4表达对缺氧/复氧H9C2心肌细胞凋亡的影响及其机制

  

  • 出版日期:2019-11-15 发布日期:2019-11-30

Effect of RNA interferring programmed cell death 4 expression on apoptosis of hypoxia/reoxygenation H9C2 cardiomyocytes and its mechanism

  • Online:2019-11-15 Published:2019-11-30

摘要: 目的 探讨RNA干扰程序性细胞死亡因子4(PDCD4)表达对缺氧/复氧(H/R)H9C2心肌细胞凋亡的影响及其机制。方法 将H9C2心肌细胞分为对照组、H/R组、H/R+NC组和H/R+siPDCD4组。采用逆转录-聚合酶链反应(RT-PCR)和蛋白质印迹法(Western blot)分别检测4组H9C2心肌细胞中PDCD4 mRNA和蛋白水平,流式细胞仪检测其凋亡率,酶联免疫吸附试验(ELISA)检测其上清液中丙二醛(MDA)、超氧化物歧化酶(SOD)、乳酸脱氢酶(LDH)水平,Western blot检测磷脂酰肌醇-3-激酶(PI3K)/蛋白激酶B(AKT)信号通路相关蛋白水平。结果H/R组H9C2心肌细胞PDCD4 mRNA和蛋白、活化的Caspase-3、Bcl-2相关X(Bax)蛋白、上清液中LDH、MDA水平及凋亡率均高于对照组,而上清液中SOD水平、H9C2心肌细胞Bcl-2和p-AKT蛋白水平均低于对照组(P<0.05)。H/R+siPDCD4组H9C2心肌细胞PDCD4 mRNA和蛋白、活化的Caspase-3、Bax蛋白、上清液中LDH、MDA水平及凋亡率均低于H/R组,而上清液中SOD、H9C2心肌细胞Bcl-2和p-AKT蛋白水平均高于H/R组(P<0.05)。结论 RNA干扰PDCD4表达可抑制H/R引起的H9C2心肌细胞凋亡,其作用机制可能与激活PI3K/AKT信号通路进而降低氧化应激反应有关。

关键词: 缺氧/复氧; 程序性细胞死亡因子4; 心肌细胞凋亡; 磷脂酰肌醇-3-激酶/蛋白激酶B信号通路; 氧化应激

Abstract: Objective To explore the effect of RNA interferring programmed cell death 4(PDCD4) expression on apoptosis of hypoxia/reoxygenation(H/R) H9C2 cardiomyocytes and its mechanism.Methods H9C2 cardiomyocytes were divided into control group,H/R group,H/R+NC group and H/R+siPDCD4 group.Levels of PDCD4 mRNA and protein of H9C2 cardiomyocytes in 4 groups were detected by reverse transcription-polymerase chain reaction(RT-PCR) and Western blotting respectively.Apoptosis rates of them were detected by flow cytometry.Levels of malondialdehyde(MDA),superoxide dismutase(SOD) and lactate dehydrogenase(LDH) in supernatant of H9C2 cardiomyocytes were measured by enzyme Linked immunosorbnent assay(ELISA).Levels of the phosphatidylinositol-3-kinase(PI3K)/protein kinase B(AKT) signaling pathway related proteins were examined by Western blotting.Results Levels of PDCD4 mRNA and protein,activated Caspase-3,Bcl-2 related X(Bax) protein,LDH,MDA and apoptosis rate of H9C2 cardiomyocytes in H/R group were higher than those in control group,while levels of SOD in supernatant and Bcl-2 and p-AKT protein in H9C2 cardiomyocytes were lower than those in control group(P<0.05).Levels of PDCD4 mRNA and protein,activated Caspase-3,Bax protein,LDH,MDA and apoptosis rate of H9C2 cardiomyocytes in H/R+siPDCD4 group were lower than those in H/R group,while levels of SOD in supernatant,Bcl-2 and p-AKT protein in H9C2 cardiomyocytes were higher than those in H/R group(P<0.05).Conclusion RNA interferring PDCD4 expression can inhibit H/R induced H9C2 cardiomyocyte apoptosis,and its mechanism may be related with the activation of PI3K/AKT signaling pathway to reduce oxidative stress.

Key words: Hypoxia reoxygenation; Programmed cell death 4; Cardiomyocyte apoptosis; Phospoinositide-3-kinase/protein kinase B signaling pathway; Oxidative stress