Gas6通过PI3K|Akt|FoxO3a通路在预适应中起心肌细胞保护作用
发布时间：2019-05-12 11:41
【摘要】：目的:初步探讨Gas6在预适应中对大鼠心肌细胞缺氧|复氧损伤的保护作用及机制。 方法:选择心肌细胞株H9C2建立实验模型,实验分为5组:①正常对照组:正常培养H9C2细胞;②缺氧复氧组:缺氧2h,再复氧24h;③预适应组:先缺氧20min,再复氧45min,随后再缺氧2h,再复氧24h;④加外源性重组人生长停滞特异性蛋白6(rhGas6)组:加入终浓度为100ng/ml的rhGas6,余同缺氧复氧组;⑤Gas6+LY294002组:加入终浓度为100ng/ml的rhGas6和终浓度为10umol/L的PI3K通路阻断剂LY294002,余同缺氧复氧组。应用流式细胞术测细胞凋亡率,测LDH活性了解心肌细胞受损程度;real time -PCR法测Gas6mRNA表达;免疫印迹法测(Western blot)测定磷酸化Akt、FoxO3a蛋白的表达。 结果:预适应组Gas6 mRNA表达高于缺氧复氧组(P〈0.05);与缺氧复氧组对比,预适应组和Gas6组LDH的活性及细胞凋亡率明显降低(P〈0.05), p-Akt和p-FoxO3a蛋白表达明显增高(P〈0.05);与Gas6组对比,Gas6+LY294002组LDH的活性及细胞凋亡率明显增加,p-Akt和p-FoxO3a蛋白表达明显减少(P〈0.05),而预适应组LDH的活性及细胞凋亡率和p-Akt、p-FoxO3蛋白表达无明显差异。 结论:Gas6通过PI3K/Akt/FoxO3a信号通路在预适应中起心肌细胞保护作用。
[Abstract]:Aim: to investigate the protective effect and mechanism of Gas6 on hypoxia / reoxygenation injury in rat cardiomyocytes. Methods: the experimental model of myocardial cell line H9C2 was established and divided into 5 groups: (1) normal control group: normal cultured H9C2 cells, (2) anoxic reoxygenation group: hypoxia for 2 hours, rereoxygenation for 24 hours, and normal control group for 24 hours. (3) preconditioning group: hypoxia for 20 min, then reoxygenation for 45 min, then hypoxia for 2 h, reoxygenation for 24 h and exogenous recombinant human growth stagnation-specific protein 6 (rhGas6) group: the rhGas6, with final concentration of 100ng/ml was added to the hypoxia-reoxygenation group; 5Gas6 LY294002 group: the addition of rhGas6 with final concentration of 100ng/ml and LY294002, a PI3K pathway blocker with final concentration of 10umol/L, was the same as anoxic reoxygenation group. The apoptosis rate was measured by flow cytometry, the activity of LDH was measured to investigate the degree of cardiomyocyte damage, the expression of Gas6mRNA was measured by; real time-PCR method, and the expression of phosphorylated Akt,FoxO3a protein was detected by (Western blot). Results: the expression of Gas6 mRNA in the preconditioning group was higher than that in the hypoxia-reoxygenation group. Compared with hypoxia and reoxygenation group, the activity of LDH and apoptosis rate in pretreatment group and Gas6 group were significantly decreased, and the expression of p-Akt and p-FoxO3a protein was significantly increased (P < 0.05). Compared with Gas6 group, the activity and apoptosis rate of LDH and the expression of p-Akt and p-FoxO3a protein in Gas6 LY294002 group were significantly increased, while the activity, apoptosis rate and apoptosis rate of LDH in preconditioned group were significantly decreased (P < 0.05). There was no significant difference in the expression of p-FoxO3 protein. Conclusion: Gas6 plays a protective role in cardiac myocytes through PI3K/Akt/FoxO3a signaling pathway.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R363
本文编号：2475362
[Abstract]:Aim: to investigate the protective effect and mechanism of Gas6 on hypoxia / reoxygenation injury in rat cardiomyocytes. Methods: the experimental model of myocardial cell line H9C2 was established and divided into 5 groups: (1) normal control group: normal cultured H9C2 cells, (2) anoxic reoxygenation group: hypoxia for 2 hours, rereoxygenation for 24 hours, and normal control group for 24 hours. (3) preconditioning group: hypoxia for 20 min, then reoxygenation for 45 min, then hypoxia for 2 h, reoxygenation for 24 h and exogenous recombinant human growth stagnation-specific protein 6 (rhGas6) group: the rhGas6, with final concentration of 100ng/ml was added to the hypoxia-reoxygenation group; 5Gas6 LY294002 group: the addition of rhGas6 with final concentration of 100ng/ml and LY294002, a PI3K pathway blocker with final concentration of 10umol/L, was the same as anoxic reoxygenation group. The apoptosis rate was measured by flow cytometry, the activity of LDH was measured to investigate the degree of cardiomyocyte damage, the expression of Gas6mRNA was measured by; real time-PCR method, and the expression of phosphorylated Akt,FoxO3a protein was detected by (Western blot). Results: the expression of Gas6 mRNA in the preconditioning group was higher than that in the hypoxia-reoxygenation group. Compared with hypoxia and reoxygenation group, the activity of LDH and apoptosis rate in pretreatment group and Gas6 group were significantly decreased, and the expression of p-Akt and p-FoxO3a protein was significantly increased (P < 0.05). Compared with Gas6 group, the activity and apoptosis rate of LDH and the expression of p-Akt and p-FoxO3a protein in Gas6 LY294002 group were significantly increased, while the activity, apoptosis rate and apoptosis rate of LDH in preconditioned group were significantly decreased (P < 0.05). There was no significant difference in the expression of p-FoxO3 protein. Conclusion: Gas6 plays a protective role in cardiac myocytes through PI3K/Akt/FoxO3a signaling pathway.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R363
【参考文献】
相关期刊论文 前2条
1 欧阳伟;第二保护窗:心肌缺血预适应的研究进展[J];国外医学(生理、病理科学与临床分册);1999年04期
2 杨彦玲,师养荣,李建龙,陈雅慧;心肌缺血再灌注损伤研究进展[J];心血管病学进展;2003年02期
，本文编号：2475362
本文链接：https://www.wllwen.com/xiyixuelunwen/2475362.html
最近更新
教材专著
