中华急诊医学杂志
中華急診醫學雜誌
중화급진의학잡지
CHINESE JOURNAL OF EMERGENCY MEDICINE
2014年
12期
1322-1326
,共5页
李莉%古正涛%刘志锋%苏磊
李莉%古正濤%劉誌鋒%囌磊
리리%고정도%류지봉%소뢰
人脐静脉内皮细胞%凋亡%线粒体%热应激%重症中暑%细胞色素C%caspase-9、caspase-3
人臍靜脈內皮細胞%凋亡%線粒體%熱應激%重癥中暑%細胞色素C%caspase-9、caspase-3
인제정맥내피세포%조망%선립체%열응격%중증중서%세포색소C%caspase-9、caspase-3
Human umbilical vein endothelial cell%Apoptosis%Mitochondrial%Heat stress%Severe heatstroke%Cytochrome C%Caspase-9%Caspase-3
目的 观察细胞色素C、caspase-9和caspase-3在热应激后人脐静脉内皮细胞的表达及相互关系,阐明热应激后人脐静脉内皮细胞凋亡的信号转导机制,探讨重症中暑所致血管内皮损害的发病机理.方法 建立人脐静脉内皮细胞热应激模型,对照组将细胞置于标准37℃、5％ CO2细胞培养箱,热应激组将细胞置于39℃、41℃、43℃细胞培养箱中进行热应激2h,热应激后继续在细胞培养箱孵育24h.电子显微镜观察人脐静脉内皮细胞(37℃、43℃)线粒体改变,应用Annexin V-FITC/PI双染色方法检测不同温度热应激后细胞凋亡率、Western blot检测细胞色素C、caspase-9、caspase-3蛋白表达.结果 电镜观察对照组(37℃)HUVEC细胞线粒体结构基本正常.热应激后(43℃)HUVEC细胞线粒体肿胀并出现结构改变;与对照组比较,39℃热应激对内皮细胞凋亡无明显影响(P＞0.05),随着热应激温度的增加人脐静脉内皮细胞凋亡明显增多(41℃17.8％,4 3℃25.6％)并且细胞色素C、caspase-9、caspase-3蛋白表达明显增加(P＜0.05).结论 线粒体通路的激活参与了热应激后人脐静脉内皮细胞凋亡的调控;血管内皮细胞凋亡可能与重症中暑的发病存在相关性.
目的 觀察細胞色素C、caspase-9和caspase-3在熱應激後人臍靜脈內皮細胞的錶達及相互關繫,闡明熱應激後人臍靜脈內皮細胞凋亡的信號轉導機製,探討重癥中暑所緻血管內皮損害的髮病機理.方法 建立人臍靜脈內皮細胞熱應激模型,對照組將細胞置于標準37℃、5％ CO2細胞培養箱,熱應激組將細胞置于39℃、41℃、43℃細胞培養箱中進行熱應激2h,熱應激後繼續在細胞培養箱孵育24h.電子顯微鏡觀察人臍靜脈內皮細胞(37℃、43℃)線粒體改變,應用Annexin V-FITC/PI雙染色方法檢測不同溫度熱應激後細胞凋亡率、Western blot檢測細胞色素C、caspase-9、caspase-3蛋白錶達.結果 電鏡觀察對照組(37℃)HUVEC細胞線粒體結構基本正常.熱應激後(43℃)HUVEC細胞線粒體腫脹併齣現結構改變;與對照組比較,39℃熱應激對內皮細胞凋亡無明顯影響(P＞0.05),隨著熱應激溫度的增加人臍靜脈內皮細胞凋亡明顯增多(41℃17.8％,4 3℃25.6％)併且細胞色素C、caspase-9、caspase-3蛋白錶達明顯增加(P＜0.05).結論 線粒體通路的激活參與瞭熱應激後人臍靜脈內皮細胞凋亡的調控;血管內皮細胞凋亡可能與重癥中暑的髮病存在相關性.
목적 관찰세포색소C、caspase-9화caspase-3재열응격후인제정맥내피세포적표체급상호관계,천명열응격후인제정맥내피세포조망적신호전도궤제,탐토중증중서소치혈관내피손해적발병궤리.방법 건립인제정맥내피세포열응격모형,대조조장세포치우표준37℃、5％ CO2세포배양상,열응격조장세포치우39℃、41℃、43℃세포배양상중진행열응격2h,열응격후계속재세포배양상부육24h.전자현미경관찰인제정맥내피세포(37℃、43℃)선립체개변,응용Annexin V-FITC/PI쌍염색방법검측불동온도열응격후세포조망솔、Western blot검측세포색소C、caspase-9、caspase-3단백표체.결과 전경관찰대조조(37℃)HUVEC세포선립체결구기본정상.열응격후(43℃)HUVEC세포선립체종창병출현결구개변;여대조조비교,39℃열응격대내피세포조망무명현영향(P＞0.05),수착열응격온도적증가인제정맥내피세포조망명현증다(41℃17.8％,4 3℃25.6％)병차세포색소C、caspase-9、caspase-3단백표체명현증가(P＜0.05).결론 선립체통로적격활삼여료열응격후인제정맥내피세포조망적조공;혈관내피세포조망가능여중증중서적발병존재상관성.
Objective To observe the expressions of cytochrome C,Caspase-9,Caspase-3 and their relationships,and investigate apoptosis signal transduction mechanism after heat stress-induction in human umbilical vein endothelial cell (HUVEC),and explore pathogenesis of vascular endothelial damage in the wake of severe heat stroke.Methods Human umbilical vein endothelial cell heat stress model was set up.Control group were incubated at 37℃,while heat stress group of cells were incubated at 39℃,41℃,and 43℃ for 2h,then all the cells were further incubated at 37℃ for 24 h.Mitochondria of human umbilical vein endothelial cell were examined with electron microscopy.Apoptosis was analyzed by flow cytometry using Annexin V-FITC/PI staining,and protein levels of cytochrome C,caspase-9,caspase-3 were analyzed by western blot.Results In the control group (37℃),the structure of mitochondrial was intact in HUVEC under transmission electron microscope.In contrast,mitochondrial swelling was found in the group of 43℃ heat stress.Compared with control group,as increasing in heat stress temperature,the rates of induced apoptosis were 17.8％ at 41℃ and 25.6％ at 43℃,and the levels of cytochrome C,Caspase-9,and caspase-3 were significantly increased (P ＜0.05).There was no obvious change at 39℃ heat stress (P ＞ 0.05).Conclusions Mitochondrial apoptosis pathway is involved in apoptosis of human umbilical vein endothelial cells in the wake of heat stress.The vascular endothelial cells apoptosis may be associated with the occurrence of severe heat stroke.
