摘要
目的:探讨caspase-3活化对热打击诱导人脐静脉内皮细胞凋亡的影响。方法:建立人脐静脉内皮细胞热打击模型,对照组将细胞置于标准37℃、5%CO2细胞培养箱,热打击组将细胞置于39℃、41℃、43℃细胞培养箱中进行热打击2 h,热打击后继续在细胞培养箱孵育24 h。使用Hoechst33258荧光染色检测细胞凋亡、Western blot检测caspase-3蛋白表达、同时检测caspase-3抑制剂Z-DEVD-FMK对热打击组细胞凋亡的影响。结果:与对照组比较,39℃热打击对内皮细胞凋亡无影响,随着热打击温度的增加人脐静脉内皮细胞凋亡明显增多、caspase-3蛋白表达明显增加,caspase-3抑制剂Z-DEVD-FMK明显抑制了热打击诱导的人脐静脉内皮细胞凋亡及caspase-3蛋白的表达。结论:caspase-3介导了热打击诱导的人脐静脉细胞的凋亡。
Objective To explore the effect of activation of caspase-3 on apoptosis of human umbilical vein endothelial cells induced by heat stress. Methods A model of human umbilical vein endothelial cell heat stress was established. The endothelial cells in the control group were incubated at 37 ℃, while the cells in the heat stress group were incubated at 39 ℃, 41 ℃, or 43 ℃ for 2 h, then all the cells were further incubated at 37 ℃ for 24 h. Apoptosis was detected using staining with Hoechst33258; protein levels of caspase-3 were analyzed by Western blot; and the effect of caspase-3 inhibitor Z-DEVD-FMK on heat stress-induced apoptosis was determined. Results As compared with the control group, with an elevation in heat stress temperature （41 ℃ to 43 ℃ ）, apoptosis was markedly increased and level of caspase-3 was significantly increased. In addition, caspase-3 inhibitor Z-DEVD-FMK significantly decreased the heat stress-induced apoptosis and levels of caspase- 3. Conclusions Caspase-3 mediates apoptosis of human umbilical vein endothelial cells can be induced by heat stress.
出处
《实用医学杂志》
CAS
北大核心
2014年第6期871-874,共4页
The Journal of Practical Medicine
基金
国家自然科学基金(编号:81071529
81101467)
广东省自然科学基金(编号:10151001002000001)
广东省科技计划项目(编号:2012B031800416)
军队十二.五医学科研基金重点项目(编号:BWS12J018)
