热刺激对体外培养的血管内皮细胞损伤及IL-6、TNF-α释放的影响  

Effect of heat stress on the injury of vascular endothelial cells and release of IL-6 and TNF-α in vitro

作　　者：潘志国[1,2] 耿焱[2] 张剑明[3] 毕媛[2] 刘云松[2] 苏磊[2] 

机构地区：[1]广州中医药大学,广州510405 [2]广州军区广州总医院全军热区创伤与组织修复重点实验室 [3]南方医科大学南方医院

出　　处：《山东医药》2012年第3期32-35,共4页Shandong Medical Journal

基　　金：广东省自然科学基金资助项目(9151001002000021)

摘　　要：目的探讨中暑发生及发展的机制。方法取对数生长期人脐静脉血管内皮细胞HUEVC,对照组将细胞置于标准37℃、5%CO2浓度培养箱中,39℃组、41℃组、43℃组分别将细胞置于39℃、41℃、43℃细胞培养箱中,均分别培养0、1、3、5、7 h。采用CCK-8法计算细胞存活率;撤除热刺激,各组更换100μL/孔完全培养基,继续培养24 h,计算细胞增殖率。采用ELISA法测定热刺激后1 h各组血浆IL-6和TNF-α水平。结果与对照组比较,39℃、41℃、43℃组各个时点细胞存活率均下降(P均<0.001),并呈时间及温度依赖关系。撤除热刺激、培养24 h时,与对照组比较,39℃组7 h及41℃组和43℃组各时点细胞增殖率均显著下降(P均<0.001),并呈时间及温度依赖关系。39℃、41℃、43℃组IL-6和TNF-α水平均明显高于对照组(P均<0.001),41℃组和43℃组较39℃组显著上升(P<0.01),43℃组和41℃组无显著性差异。结论热刺激对血管内皮细胞具有细胞毒效应,可抑制血管内皮细胞增殖,并促进细胞因子IL-6和TNF-α的释放,且呈温度及时间依赖性。本研究结果为中暑的发生及发展机制研究奠定了实验基础。Objective To investigate the mechanism of the pathogenesy of heatstroke. Methods Human umbilical endothelial cell line HUEVC in exponential phase were divided into 4 groups randomly, the control group was cultured in standard incubator of 37 ℃, 5% CO2 , the groups of 39 23,41 23,43 23 were cultured in incubator of 39 23,41 23,43 23, the culture time was 0, 1,3, 5, 7 h. CCK-8 assay was used to investigate the difference of cell survival rate and 24 h proliferation rate （ after the removal of heat sress and complete medium changes of 100 μL each hollow）. ELISA assays was used to investigate the level of IL-6 and TNF-α. Results In comparison with control group, the cell vitality of 39 ℃, 41 23,43 23 groups were significantly decreased （ all P 〈 0. 001 ）, and appeared time and thermal gradient dependencies. When removed heat stress and cultured for 24 h, the proliferation rate of 7 h phase in 39 23 group and each phase of 4 1℃ and 43 23 groups were significantly decreased in comparison with control group （ all P 〈 0.001 ）, which also showed time and thermal gradient dependencies. The expression level of IL-6 and TNF-α in 39 ℃, 41 23, 43 23 groups were significantly up-regulated in comparison with control group （all P 〈0.001 ）. The cells of 41 23 and 43 23 groups released more IL-6 and TNF-α than cells of 39 23 group （P 〈 0.01 ）, and there was no significant difference in IL-6 and TNF-α released between 41 23 and 43 23 groups. Conclusion Heat stress has cytotoxic effect on vascular endothelial cell, and can inhibit proliferation ability of vascular endothelial cell, can promote the IL-6 and TNF-α release of vascular endothelial ceil, all in time and thermal gradient-dependent manner. This study may help to understand the mechanism of the pathogenesy of heatstroke.

关 键 词：热刺激 血管内皮细胞 细胞毒 细胞增殖 细胞因子 

分 类 号：R363[医药卫生—病理学] R594.1[医药卫生—基础医学]