热应激对大鼠肠上皮细胞增殖和凋亡的影响  

Effects of heat stress on proliferation and apoptosis of rat intestinal epithelial cells

作　　者：殷启润 李冰 熊永洁 贺绍君[1] YIN Qirun;LI Bing;XIONG Yongjie;HE Shaojun(College of Animal Science,Anhui Science and Technology University,Fengyang 223100,China)

机构地区：[1]安徽科技学院动物科学学院,安徽凤阳233100

出　　处：《畜牧与兽医》2021年第12期60-64,共5页Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金青年基金(31702306);安徽省高校优秀拔尖人才培育项目(gxyq2020038);安徽省科技厅自然科学基金(KJ2019A0082)。

摘　　要：试验旨在探究热应激对IEC-6细胞增殖、凋亡、周期、凋亡及紧密连接蛋白表达的影响。试验采用体外细胞培养热应激模型,分别设置对照组(37℃)与热应激组(41℃)对细胞进行处理,每组3个重复,CCK-8法检测IEC-6细胞活性,流式细胞术检测IEC-6细胞凋亡率及周期,Western blot法检测凋亡相关蛋白(Bax、Bcl-2和Caspase-3)以及紧密连接蛋白(Occludin和Claudin-1)表达水平。结果显示,与对照组相比,热应激处理组的IEC-6细胞活力随温度升高而下降,细胞凋亡率显著上升(P<0.05);细胞周期的G0/G1期阻滞;S期以及G2/M期峰值显著低于对照组(P<0.05);热应激处理后细胞的增殖指数显著降低(P<0.05);Western blot结果显示,热应激极显著增强Bax和Caspase-3蛋白表达量(P<0.01),极显著抑制Bcl-2、Occludin与Claudin-1蛋白表达水平(P<0.01)。结果表明,热应激可以显著降低IEC-6细胞的活性,促进细胞凋亡的发生,抑制细胞增殖,上调细胞内凋亡蛋白的表达,抑制细胞中紧密连接蛋白表达。The aim of this study was to investigate the effects of heat stress on the apoptosis and tight junction protein expression of IEC-6 cells. In this experiment, the heat stress model of cell culture in vitro was used, and the control group(37 ℃) and the heat stress group(41 ℃) were formed to treat the tested cells.There were three repeats in each group.CCK-8 assay was used to detect IEC-6 cell activity;flow cytometry was used to determine the apoptosis rate and cycle of IEC-6 cells, and Westernblot was used to identify the expression levels of Bax, Bcl-2, Caspase-3, Occludin and Claudin-1 proteins. The results were that, compared with the control group, IEC-6 cell viability in the heat stress group decreased as temperature increased;the apoptosis rate of IEC-6 cells increased significantly(P<0.05);the G0/G1 of the IEC-6 cell cycle retarded;the peak values of the S period and the G2/M period weresignificantly lower than that of the control group(P<0.05);and the proliferation index of the IEC-6 cells decreased significantly after the heat stress treatment(P<0.05). The results of Western blot showedthat thermal stress significantly enhanced the expression of the Bax and Caspase-3 proteins(P<0.01), and the expression of Bcl-2, Occludin and Claudin-1 proteins was significantly inhibited(P<0.01). To conclude, heat stress significantly reduced the activity of IEC-6 cells, promoted their apoptosis, suppressed their proliferation, upregulated the apoptotic protein expression in cells, and inhibitedthe expression of thetight junction protein in the cells.

关 键 词：热应激 IEC-6细胞 凋亡 紧密连接蛋白 

分 类 号：S826.3[农业科学—畜牧学]