磷脂酰肌醇3-激酶γ抑制剂AS605240对系统性红斑狼疮患者外周血T细胞增殖、黏附和趋化的调控研究

Study on effect of PI3Kγ inhibitor AS605240 proliferation,adhesion and chemotaxis of peripheral blood T cells from patients with systemic lupus erythematous

目的:研究磷脂酰肌醇3-激酶γ(PI3Kγ)抑制剂AS605240对系统性红斑狼疮(SLE)患者外周血T淋巴细胞增殖、黏附和趋化的影响,并初步探讨其机制。方法:搜集SLE患者35例,健康人17例为正常组,外周血T细胞分离采用Rosettsep T细胞提取试剂盒;四甲基偶氮唑蓝(MTT)比色法检测细胞增殖,Transwell小室检测T细胞趋化性,丝氨酸/苏氨酸蛋白激酶(AKT)活性检测采用免疫印迹法。结果:AS605240显著抑制SLE患者外周血T细胞体外增殖、黏附活性,并呈浓度依赖关系,当AS605240浓度达到10^(-5)mol/L时差异有统计学意义(P<0.01),AS605240显著抑制SLE患者外周血T细胞体外趋化活性(P<0.01);SLE患者外周血T细胞AKT磷酸化水平显著高于正常组(P<0.05),AS605240处理组T细胞的AKT活性显著降低(P<0.05)。结论:P13Kγ抑制剂AS605240显著降低SLE患者外周血T细胞增殖、趋化和黏附活性,与抑制AKT异常活化相关。

Objective： To investigate the effect of PI3Kγ inhibitor AS605240 on the proliferation, adhesion and chemotaxis of the T cells from the patients with systemic lupus erythematous（SLE）. Methods： T cells were isolated from the peripheral blood samples of 35 SLE patients and 17 healthy adults as controls by Rosettsep T cell purification kits. The proliferation of T cells and chemotaxis were examined by MTF and Transwell chambers, respectively. Western blot was used to detect AKT activity. Results： AS605240 significantly inhibited T cells proliferation and adhesion activities in dose-dependent manner, when the concentration of AS605240 reached 10-5 mol/L, the difference was statistically significant （P〈0.01）. The ehemotactic activities of T cells from SLE patients were obviously inhibited （P〈0.01）. Compared with the control group, the expression of phosphorylated Akt were higher and could be suppressed by AS605240 （P〈0.05）. Conclusion： PI3Kγ inhibitor AS605240 significantly reduce the activities of the proliferation, chemotaxis and adhesion of the peripheral blood T cells in SLE patients, the mechanism may be related to the suppression of abnormal activation of AKT.