线粒体生物合成的改变对高糖诱导的活性氧类生成的影响

摘要

Objective To study the effect of PGC-1α on vascular endothelial cells mitochondrial biogenesis and the influence of biogenesis on glucose induced ROS production.Methods Primary culture of BAECs and HUVECs were divided into four groups,each group was cultured in normal glucose solution and in high glucose solution(concentration:5,30 mmol/L),and the intracellular ROS concentration was detected.Results Respective covariance analysis for BAECs and HUVECs showed that,the difference were statistically significant among the different groups (P ＜ 0.01),different glucose level had significant impact on the concentration of ROS (P ＜ 0.05) ; analysis on BAECs showed that the ROS concentration of PGC-1 α transfected group on normal glucose level was significantly lower than that of the control group and empty plasmid group (P ＜ 0.01),on high glucose level was lower than that in the control group and the empty plasmid group (P ＜ 0.05) ;the ROS concentration of PGC-1 α siRNA transfected group on normal glucose level was significantly higher than that of the control group(P ＜ 0.01),no statistically significant difference in comparison with the empty plasmid group(P ＞ 0.05) ; analysis on the HUVECs showed that the ROS concentration of PGC-1 α infected group was lower than the empty virus Ad group and the control group on normal and high glucose level(P ＜ 0.01);the ROS concentration of PGC-1 α siRNA transfected group on normal glucose level was significantly higher than the control group(P ＜0.01),no significant difference between Ad group and empty virus group(P ＞0.05).Conclusion On high glucose level,more ROS is generated by vascular endothelial cells than on the normal glucose level,but after PGC-1α protein is over expressed,the intracellular ROS concentration is obviously decreased; otherwise the ROS concentration is significantly increased.%目的 探讨过氧化物酶体增殖激活受体γ辅助激活因子1α(PGC-1α)对血管内皮细胞内线粒体生物合成的影响,明确线粒体的生物合成对葡萄糖诱导的活性氧类(ROS)生成的影响.方法 将原代培养的人脐带静脉内皮细胞(HUVECs)及小牛主动脉内皮细胞(BAECs)分成四大组,并分别将四组细胞模拟人体正常糖及高糖环境,浓度为5、30 mmol/L进行培养,测量细胞内ROS浓度.结果 对BAECs和HUVECs分别进行协方差分析结果显示,在两种细胞中,不同组别之间差异均有统计学意义(P＜0.01),不同糖水平对ROS浓度有影响(P＜0.05)；对BAECs单独进行分析发现,PGC-1α抑制表达组的ROS浓度在正常糖水平下显著低于对照组及空质粒组(P＜0.01),高糖水平下也低于对照组及空质粒组(P＜0.05)；PGC-1α过度表达组的ROS浓度正常糖水平显著高于对照组(P＜0.01),与空质粒组相比差异无统计学意义(P＞0.05)；对HUVECs分析发现,PGC-1α抑制表达组ROS浓度在正常糖水平和高糖水平下均低于空病毒Ad组和对照组(P＜0.01)；PGC-1α siRNA转染组的ROS浓度正常糖水平显著高于对照组(P＜0.Ol),与空质粒组差异无统计学意义(P＞0.05).结论 高糖组较正常糖组血管内皮细胞产生更多ROS,过度表达PGC-1α后,细胞内ROS浓度显著降低；反之,ROS浓度显著升高.