人参皂苷Rb_1对小鼠T淋巴细胞体外活化、增殖及凋亡的影响

Effects of ginsenoside Rb_1 on activation, proliferation,and apoptosis of murine T lymphocytes in vitro

目的研究人参皂苷Rb1对小鼠T淋巴细胞体外活化及增殖的影响,并对其免疫调节作用进行初步探讨。方法分离制备小鼠淋巴结细胞悬液;以双色荧光抗体染色技术结合流式细胞术检测在刀豆蛋白A(ConA)的刺激下,人参皂苷Rb1对早期活化标志CD3/CD69和调节T淋巴细胞标志CD4/CD25表达的影响;用羧基荧光素乙酰乙酸(CFDA-SE)染色结合流式细胞术和MTT法检测在ConA的刺激下,人参皂苷Rb1对T淋巴细胞增殖的影响;DIOC/PI双染技术检测在H2O2作用下,人参皂苷Rb1对淋巴细胞凋亡进程的影响。结果终浓度为5、10、20μmol/L的人参皂苷Rb1对ConA刺激的调节T淋巴细胞CD4+/CD25+的表达具有促进作用,而CD3+/CD69+的表达明显下调(P<0.01);对ConA刺激的T淋巴细胞增殖具有明显的抑制作用(P<0.05),且呈剂量依赖性;人参皂苷Rb1显著抑制H2O2诱导的淋巴细胞的凋亡进程(P<0.05)。结论人参皂苷Rb1对小鼠淋巴细胞的体外活化及增殖均具有明显的抑制作用,是一种潜在的免疫抑制剂。

Objective To investigate the effects of ginsenoside Rb1 on activation and proliferation of murine T lymphocytes in vitro and to elucidate the mechanism of the immunosuppressive effect of ginsenoside Rb1.Methods Cell suspensions were prepared from murine lymph nodes.T lymphocytes were treated with different concentrations of ginsenoside Rb1 and stimulated with polyclonal activator concanavalin（ConA）.Fluorescence conjugated monoclonal antibodies and flow cytometry were used to detect the expression of CD3/CD69 and CD4/CD25.After the staining with CFDA-SE,T lymphocytes were stimulated with polyclonal activator ConA.The proliferation of T lymphocytes,after stimulated by ConA,was detected using the method of MTT.The distribution of the cell apoptosis was analyzed by staining both DIOC and PI.Results In a dose-dependent manner,ginsenoside Rb1（5,10,and 20 μmol/L） could significantly inhibit T lymphocytes activation index（P0.01） stimulated by ConA and proliferation index（P0.05） stimulated by ConA.Ginsenoside Rb1 could also reduce the apoptosis of T lymphocytes stimulated by H2O2.Conclusion Ginsenoside Rb1 can effectively inhibit the activation and proliferation of murine T lymphocytes,and ginsenoside Rb1 is a potential effective immunoinhibitory agent.