肿瘤坏死因子α对肠上皮细胞株HT-29的核转录因子κB信号通路的激活作用研究

Study of tumor necrosis factor-α on activation of the nuclear factor-κB pathway in intestinal epithelial cell line HT-29

目的:研究肿瘤坏死因子α(TNF-α)对肠上皮细胞株HT-29的核转录因子κB(NF-κB)信号通路的激活作用及鼠尾草酚对TNF-α诱导HT-29的NF-κB激活的影响。方法:将HT-29细胞分为空白对照组、TNF-α组和低、中、高剂量鼠尾草酚组。用CCK-8法检测HT-29细胞活力;Western-blot方法检测IκBα、Pi-IκBα和PiIKKα/β蛋白表达;用NF-κB转录活性试剂盒检测HT-29细胞NF-κB转录活性。结果 :实验各组的HT-29细胞活力比较无显著性差异(P>0.05);TNF-α刺激可诱导HT-29的IκBα磷酸化及降解;鼠尾草酚能使TNF-α诱导HT-29的IKKα/β和IκBα磷酸化显著受抑制(均P<0.05),并且可显著降低NF-κB的转录活性(P<0.05)。结论:TNF-α可激活HT-29的NF-κB经典信号通路,鼠尾草酚对TNF-α诱导的NF-κB激活有抑制作用。

Objective：To study the effect of tumor necrosis factor（TNF）-α on activation of nuclear faetor（NF） -κB pathway in intestinal epithelial cell line HT-29 as well as effect of carnosol on NF-κB pathway in TNF-α-stimulated HT-29.Methods：HT-29 cells were divided into five groups：control group,TNF-α group, and low-,mid-,high-dose carnosol groups.Cell counting kit-8 （CCK-8）was used for analysis of HT-29 viability.Expression of IκBα,Pi-IκBα and Pi-IKKα/β in HT-29 were evaluated by Western-blot. NF-κB-DNA binding activation of HT-29 was analyzed with the TransAM NF-κB Family Kit.Results：There were no significant differences on cell viability in five groups （P〉0.05）.TNF-α could induce IκBα phosphorylation and degradation in HT-29.Carnosol significantly inhibited IKKα/β and IκBα phosphorylation and NF-κB-DNA binding activation in TNF-α-stimulated HT-29 （P〈0.05）.Conclusion：TNF-α induces activation of NF-κB classic pathway in HT-29 and carnosol inhibits TNF-α-indueed activation of NF-κB in HT-29.Carnosol can be used in inflammatory bowel disease （IBD）due to its anti-inflammatory properties targeting intestinal epithelial cells.