摘要
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、p AKt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 20μmol/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、p AKt、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。
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 and15 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 PI3 K,p AKt,mTOR,BCL-2,BAX and caspase-3. Besides,the PBMCs of SLE patients were treated with HCQ 25 mg/L and the PI3 K/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,p AKt,mTOR and BCL-2 of HCQ group were significantly increased while the expression of BAX and caspase-3 decreased significantly（ P〈0. 05）. The PI3 K/Akt pathway inhibitor LY294002 could block the PBMCs apoptosis of SLE patients. Conclusion： Hydroxychloroquine can promote the PBMCs apoptosis of SLE patients by PI3 K/Akt signaling pathways.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2018年第1期99-103,共5页
Chinese Journal of Immunology
基金
湖北省自然科学基金项目(2012FFA061)
关键词
羟氯喹
系统性红斑狼疮
外周血单个核细胞
凋亡
PI3K/AKT信号通路
Hydroxychloroquine
Systemic lupus erythematosus
Peripheral blood mononuclear cells
Cell apoptosis
PI3K/Akt signaling pathway
