摘要
目的 :观察热休克预处理对肠上皮细胞 (IEC 6 )缺氧再给氧损伤的影响 ,并探讨热休克蛋白 70(HSP70 )的细胞保护作用。方法 :体外培养 IEC 6细胞 ,分为正常对照组、单纯缺氧再给氧组及热休克预处理组 ,观察缺氧再给氧后各组细胞 HSP70及 Bcl 2表达及细胞活力、乳酸脱氢酶 (L DH)漏出情况 ;用流式细胞仪检测细胞凋亡情况。结果 :热休克预处理可诱导 HSP70产生 (P <0 .0 1) ,显著增加缺氧再给氧处理后IEC 6细胞活力 (P<0 .0 5 ) ,使 L DH漏出减少 (P<0 .0 5 ) ,凋亡相关基因 Bcl 2表达明显增加 (P<0 .0 1) ,细胞凋亡率明显降低 (P<0 .0 1)。结论 :热休克预处理可能通过诱导 HSP70表达来减轻肠上皮细胞缺氧再给氧损伤 ;增加细胞 Bcl 2基因表达并抑制肠上皮细胞缺氧再给氧后细胞凋亡可能是其保护作用机制之一。
Objective:To study the mechanism and effect of heat shock pretreatment on hypoxiareoxygenation induced injury of intestinal epithelial cells.Methods:Cultured intestinal epithelial cell6(IEC6) cells were divided into three groups:control group (C),hypoxiareoxygenation group (HR) and heat shock pretreated group (HS+HR).After 1 hour of hypoxia followed by 1 hour of reoxygenation,immunohistochemistry was used to determine the expression of heat shock protein 70 (HSP70) and Bcl2 gene in IEC6 cells .Cell viability and lactate dehydrogenase leakage were analysed.The cell apoptosis was evaluated with flow cytometry using Annexin V Flous staining.Results:Heat shock pretreatment could improve cell viability( P <0 05),and reduce the lactate dehydrogenase leakage from IEC6 cells induced by hypoxia reoxygenation ( P <0 05).The cell apoptosis ratio in HS+HR group was significantly less than in HR group ( P <0 01).These were accompanied by an increase in the expression of HSP70( P <0 01) and Bcl2 gene( P <0 01).Conclusions:Heat shock pretreatment seems to provide protection for IEC6 cells against hypoxiareoxygenation induced injury by enhancing the expression of HSP70,and the mechanism is possibly related to the increase in the expression of Bcl2 gene and the inhibition of cell apoptosis.
出处
《中国危重病急救医学》
CAS
CSCD
2002年第5期265-268,共4页
Chinese Critical Care Medicine
基金
"973"国家重点基础研究发展规划项目(No.G19990 5 42 0 2)
关键词
热休克蛋白
凋亡
缺氧-再给氧
肠上皮细胞
热休克预处理
heat shock protein
apoptosis
hypoxiareoxygenation
intestinal epithelial cell
heat shock pretreatment
