雷帕霉素对小鼠T淋巴细胞的体外作用

Effects of rapamycin on proliferation,function and cell-cycle of mouse T lymphocytes in vitro

目的研究雷帕霉素对小鼠T淋巴细胞体外的增殖、白细胞介素2(IL-2)表达及细胞周期的影响。方法将C57BL/6小鼠脾脏制备成脾单个核细胞,使用免疫磁珠法分选出小鼠T淋巴细胞。T细胞分别加入0、10、20、30和40ng/ml梯度浓度的雷帕霉素培养液培养72h后,采用MTT法及ELISA法测定各组细胞的增殖情况及IL-2表达情况;用流式细胞仪测定40ng/ml雷帕霉素浓度组及对照组(0ng/ml)细胞周期的变化情况。结果用雷帕霉素培养液培养细胞时,细胞增殖及IL-2的表达均受到抑制(P<0.01)。随药物浓度增加,细胞增殖及IL-2表达受抑制越明显(P<0.01);细胞周期分析显示加入雷帕霉素培养的T淋巴细胞与未使用药物刺激者相比,G0G1期细胞的比例升高(P<0.01),S期降低(P<0.01)。结论在体外雷帕霉素对小鼠T细胞的增殖、IL-2的表达起抑制作用,且随浓度增加,抑制作用增强;雷帕霉素将小鼠T细胞抑制在G0G1期。

Objective To investigate the effects of rapamycin on proliferation, levels of interleukin-2 （IL-2） and cell-cycle of mouse T lymphocytes in vitro. Methods Mononuclearcell from C57BL/6 mouse＇s spleen was prepared and the memory T cells were separated by Magnetic Cell Sorting System. Various concentrations of repamycin （0, 10, 20, 30 and 40 ng/ml） were used to culture the memory T cells for 72 hours. The activity of cells and levels of IL-2 were measured by methylthiazlyltetrazolium （MTT） assay. Results Proliferation of memory T cells and expression of IL-2 were suppressed in a dose-dependent manner by rapamycin medium when cultured with rapamycin （P〈0. 01）. The cell population increseased in G0G1 phase （P〈0. 01） and decreased in S phase （P〈0. 01）. Conclusion In vitro, rapamycin is a potential inhibitor that blocks the proliferation of memory T cells in Go G1 phase,and the expression of IL-2.