IL-17调控PI3K/AKT/FAS/FASL信号通路抑制Hep-2细胞凋亡

IL-17 inhibits apoptosis of Hep-2 cell lines through modulating PI3K/AKT/FAS/FASL signaling pathway

目的研究白介素-17(IL-17)对Hep-2细胞凋亡的影响,阐明IL-17通过PI3K/AKT信号通路在抑制Fas/Fas L信号通路从而抑制Hep-2细胞凋亡的部分机制。方法以正常Hep-2细胞为对照组,50 ng/ml 200-17(IL-17刺激剂)为实验组,流式细胞仪检测200-17对Hep-2细胞凋亡率的影响。以Hep-2细胞为对照组,不同浓度200-17(1、50、100ng/ml)为实验组,Western blot法检测200-17作用6 h后各组Hep-2细胞中PI3K、p-PI3K、AKT、p-AKT、Fas、Fas L蛋白的表达。结果 Annexin V-FITC/PI细胞凋亡检测结果显示,对照组和实验组凋亡率逐渐增高,与对照组比较,加50 ng/ml 200-17作用6、12、24 h后,Hep-2凋亡率均降低,200-17作用6 h后与对照组比,实验组Hep-2细胞凋亡率降低最明显。Western blot结果显示不同浓度(0、1、50、100 ng/ml)的200-17刺激Hep-2细胞后,随着200-17浓度的升高,Hep-2细胞中p-PI3K、p-AKT蛋白的表达逐渐增加,差异有统计学意义(P <0. 01); PI3K、AKT蛋白表达差异无统计学意义。与对照组比较,1、50、100 ng/ml组200-17刺激6 h后Hep-2细胞内Fas、Fas L蛋白表达降低,差异有统计学意义(P <0. 01)。结论 IL-17可能通过PI3K/AKT/Fas/Fas L信号通路抑制Hep-2细胞凋亡。

Objective To investigate the effect of interleukin-17(IL-17)on the apoptosis of Hep-2 cells and to elucidate the mechanism by which IL-17 inhibits the apoptosis of Hep-2 cells by inhibiting the Fas/FasL signaling pathway through the PI3K/AKT signaling pathway.Methods Normal Hep-2 cells were used as the control group,and 50 ng/ml 200-17(IL-17 stimulant)was used as the experimental group.The effect of 200-17 on the apoptosis rate of Hep-2 cells was detected by flow cytometry.Hep-2 cells were grouped into control group,1 ng/ml 200-17 group,50 ng/ml 200-17 group and 100 ng/ml 200-17 group.Western blot was used to detect the expression of PI3K,p-PI3K,AKT,p-AKT,Fas and FasL in Hep-2 cells after 6 h stimulation with different concentrations of 200-17.Results Annexin V-FITC/PI cell apoptosis assay results showed that the apoptosis rate of Hep-2 cells in the control and experimental groups increased gradually.Compared with the control group,the apoptosis rate of Hep-2 decreased after adding 50 ng/ml 200-17 for 6 h,12 h,and 24 h.After 6 hour treatment with 200-17,the apoptosis rate of Hep-2 cells were significantly lower than that of the control group.The results of Western blot showed that the expressions of p-PI3K and p-AKT protein in Hep-2 cells were significantly increased with the increase of IL-17 concentration(P<0.01).The expression of PI3K and AKT had no significant change in each group.Compared with the control group,the expressions of Fas and FasL proteins in 1 ng/ml group,50 ng/ml group and 100 ng/ml group were significantly decreased(P<0.01).Conclusion IL-17 may inhibit the apoptosis of Hep-2 cells through modulating PI3K/AKT/FAS/FASL signaling pathway.