论文部分内容阅读
目的探讨热休克转录因子1(HSF1)对过氧化氢(H2O2)刺激后心脏微血管内皮细胞内活性氧簇(ROS)水平和细胞凋亡的影响。方法体外培养大鼠心脏微血管内皮细胞,分别单独转染HSF1质粒、凋亡信号调节激酶1(ASK1)质粒或共转染HSF1及ASK1质粒。转染48h后用1mmol/LH2O2刺激细胞30min,检测并比较各组细胞凋亡情况和胞内ROS水平。结果 (1)H2O2刺激后,各组细胞凋亡较刺激前明显增加(P<0.05);且在相同刺激条件下,HSF1转染组细胞凋亡明显少于对照组(P<0.05),ASK1转染组明显多于对照组(P<0.05),而共转染组与ASK1转染组相比明显减少(P<0.05)。(2)H2O2刺激后,各组细胞内ROS水平较刺激前都显著升高(P<0.05);且在相同刺激条件下,各组细胞内ROS水平较刺激前ROS水平增高的幅度:HSF1转染组明显低于对照组(P<0.05),ASK1转染组和共转染组与对照组无明显差异,而共转染组与HSF1转染组相比有升高趋势,与ASK1转染组相比有降低趋势。结论 HSF1可以通过抑制细胞内ROS产生对抗氧化应激诱导的细胞凋亡,保护心脏微血管内皮细胞。ASK1可能干扰HSF1的保护作用。
Objective To investigate the effect of heat shock transcription factor 1 (HSF1) on the level of reactive oxygen species (ROS) and apoptosis in cardiac microvascular endothelial cells stimulated by hydrogen peroxide (H2O2). Methods Rat cardiac microvascular endothelial cells were cultured in vitro and transfected with HSF1 plasmid, Apoptotic Signal-Regulating Kinase 1 (ASK1) plasmid or co-transfected with HSF1 and ASK1 plasmids respectively. 48h after transfection with 1mmol / LH2O2 stimulated cells for 30min, detect and compare the apoptosis of each group and intracellular ROS levels. Results (1) After H2O2 stimulation, the apoptosis in each group was significantly increased (P <0.05), and under the same stimulation, the apoptosis of HSF1 transfection group was significantly less than that of the control group (P <0.05) Transfection group was significantly more than the control group (P <0.05), while the co-transfected group and ASK1 transfected group was significantly reduced (P <0.05). (2) After H2O2 stimulation, intracellular ROS levels in each group were significantly increased compared with that before stimulation (P <0.05); and under the same stimulation conditions, the intracellular ROS levels in each group increased more than the pre-stimulation level of ROS: HSF1 (P <0.05). There was no significant difference between the ASK1 transfection group and the co-transfection group and the control group, while the transfection group compared with the HSF1 transfection group had a rising trend, and ASK1 transfection Compared to the group there is a downward trend. Conclusion HSF1 protects cardiac microvascular endothelial cells against oxidative stress-induced apoptosis by inhibiting intracellular ROS production. ASK1 may interfere with the protective effect of HSF1.