羟氯喹通过PI3K/Akt通路对系统性红斑狼疮外周血单个核细胞凋亡的作用研究

目的：探讨羟氯喹对SLE患者外周血单个核细胞的凋亡作用及其相关机制。方法：对30例活动期SLE患者及15例正常健康人抽血分离PBMCs细胞进行培养，分为正常对照组、SLE组、羟氯喹5 mg/L、羟氯喹25 mg/L组，MTT法检测细胞生长抑制，采用Annexin V/PI流式细胞仪细胞检测凋亡率，Western blot方法检测BAX、BCL-2、PI3K、pAKt及mTOR等相关蛋白的表达影响。同时加入羟氯喹HCQ 25 mg/L和PI3K/AKT通路抑制剂LY294002 20 μmol/L，作用SLE患者的PBMCs细胞48 h，检测PBMCs细胞生长抑制和凋亡率。结果：SLE组比正常对照组PBMCs细胞生长抑制率和凋亡率显著升高（P<0.05）；与SLE组相比，羟氯喹5 mg/L和25 mg/L组细胞生长抑制率和凋亡率显著升高（P<0.05）。羟氯喹组与SLE组比较，PI3K、pAKt、mTOR、bcl-2的表达明显下降，差异有统计学意义（P<0.05），bax和caspase-3的表达增加，差异有统计学意义（P<0.05）。PI3K/AKT抑制剂 LY294002能够阻断羟氯喹导致的SLE患者PBMCs细胞凋亡。结论：羟氯喹能够通过PI3K/Akt信号通路促进SLE患者体外PBMCs的凋亡。

Effects of Hydroxychloroquine on cell apoptosis in peripheral blood mononuclear cells of systemic lupus erythematosus by PI3K/Akt signaling pathways

Objective:To investigate the effects of Hydroxychloroquine on cell apoptosis in peripheral blood mononuclear cells of systemic lupus erythematosus and its mechanisms.Methods: The peripheral blood mononuclear cells of 30 active SLE patients and 15 healthy persons were separated for cell culture.There were four groups control group,SLE group and HCQ 5 mg/L and HCQ 25 mg/L group.MTT was used to measure the inhibitory effect.Annexin V/PI flow cytometry was performed to analyze cell apoptosis.Western blot was used to evaluate the expressions of PI3K,pAKt,mTOR,BCL-2,BAX and caspase-3.Besides,the PBMCs of SLE patients were treated with HCQ 25 mg/L and the PI3K/Akt pathway inhibitor LY294002 20 μmol/L and its cell growth inhibition and apoptosis were observed.Results: Compared with the control group,the cell growth inhibition and apoptosis rate of SLE patients group were significantly increased（P<0.05）;while the cell growth inhibition and apoptosis rate of HCQ 5 mg/L and 25 mg/L were increased significantly than the SLE patients group（P<0.05）.Compared with SLE patients group,the expression levels of PI3K,pAKt,mTOR and BCL-2 of HCQ group were significantly increased while the expression of BAX and caspase-3 decreased significantly (P<0.05).The PI3K/Akt pathway inhibitor LY294002 could block the PBMCs apoptosis of SLE patients.Conclusion: Hydroxychloroquine can promote the PBMCs apoptosis of SLE patients by PI3K/Akt signaling pathways.