诱导HSP70减轻肠上皮细胞缺氧/再给氧的损伤  

Effect of inducible HSP70 on hypoxia/reoxygenation induced injury of intestinal epithelial cell in vitro

作　　者：袁志强[1] 李晓鲁[1] 彭毅志[1] 

机构地区：[1]中国人民解放军第三军医大学西南医院烧伤研究所,重庆市400038

出　　处：《世界华人消化杂志》2002年第7期800-805,共6页World Chinese Journal of Digestology

基　　金：国家重点基础研究发展规划项目;(G1999054202)

摘　　要：目的:观察热休克预处理诱导热休克蛋白70(HSP70)对肠上皮细胞(IEC-6)缺氧/再给氧损伤的影响,并探讨 HSP70的细胞保护作用。方法:体外培养IEC-6细胞,随机分为正常对照 (C)、单纯缺氧/再给氧(H/R)及热休克预处理组 (HS+H/R),免疫组化检测缺氧/再给氧后各组细胞 HSP70表达程度,MTT法分析细胞活力,并检测细胞 LDH漏出,TUNEL法检测细胞凋亡情况。结果:与C组及H/R组比较,热休克预处理可明显诱导HSP70的产生(0.93±0.07 vs 0.20±0.04,0.39± 0.08,P<0.01),热休克预处理可显著增加IEC-6细胞缺氧/再给氧处理后细胞活力(0.27±0.04 vs 0.24± 0.02,P<0.05),LDH漏出减少(1021.29±207.06 vs 1419.90±393.07,P<0.05),细胞凋亡率也明显降低 (7.8±1.6％ vs 12.3±2.9％,p<0.05)。结论:通过热休克预处理诱导HSP70表达,可明显减轻肠上皮细胞缺氧/再给氧损伤.防止肠上皮细胞缺氧 /再给氧后细胞凋亡可能是其保护作用机制之一。AIM: To study the mechanism and effect of inducible heat shock protein 70 (HSP70) on hypoxia/ reoxygenation induced injury of intestinal epithelial cell. METHODS: Cultured IEC-6 cells were divided into three groups: Control group (C), hypoxia/reoxygenation group (H/R) and heat shock pretreated group (HS+H/R). After 1 hour of hypoxia followed by 1 hour of reoxygenation, Immunohistochemical staining was used to determine the expression of HSP70 in IEC-6 cells. Cell viability was measured by MTT assay and lactate dehydrogenase leakage were analyzed, and the cell apoptosis was evaluated with TUNEL staining. RESULTS: Heat shock pretreatment could improve cell viability(0.27±0.04 vs 0.24±0.02, P<0.05), and reduce the lactate dehydrogenase leakage(1021.29±207.06 vs 1419.90±393.07, P<0.05)from IEC-6 cells induced by hypoxia /reoxygenation. The cell apoptosis ratio in HS+H/R group was significantly less than in H/R group (7.8±1.6％ vs 12.3±2.9％, P<0.05). These were accompanied by an increase in the expression of HSP70 (0.93±0.07 vs 0.20± 0.04, 0.39±0.08, P<0.01). CONCLUSIONS: Heat shock pretreatment seems to provide protection for IEC-6 cells against hypoxia/reoxygenation induced injury by inducing the expression of HSP70, and the mechanism is possibly related to the inhibition of cell apoptosis.

关 键 词：诱导 HSP70 肠上皮细胞 缺氧 再给氧 损伤 

分 类 号：R574[医药卫生—消化系统]