摘要
目的研究白藜芦醇对小鼠CD4+T细胞增殖、凋亡和亚群的影响。方法 Balb/c鼠(SPF,雌性,9～10周龄)脾脏单个核细胞(SMC)与不同浓度(0、20、40、80 mmol/L)白藜芦醇共培养30 min,加离子霉素和PMA共培养1 h后,再加蛋白质转运阻断剂Golgiplug继续共培养4～6 h,收集细胞采用胞内细胞因子染色法结合流式细胞术检测IL-4和IFN-γ情况。免疫磁珠法从SMC分选CD4+T细胞。CD4+T细胞与不同浓度(0、10、20、40 mmol/L)白藜芦醇、抗CD3/抗CD28抗体和IL-2共培养,4 d后收集细胞采用流式细胞术检测增殖情况;CD4+T细胞与不同浓度(0、10、20、40 mmol/L)白藜芦醇共同孵育36 h后,采用流式细胞术检测其凋亡情况。结果白藜芦醇(≥20 mmol/L)可显著抑制CD4+T细胞增殖(P<0.05),抑制CD4+T淋巴细胞IFN-γ表达(P<0.05),其抑制作用均呈剂量依赖关系。白藜芦醇(≥10 mmol/L)可显著诱导CD4+T淋巴细胞凋亡(P<0.01),其作用呈剂量依赖关系。结论白藜芦醇具有抑制小鼠CD4+T细胞的增生、抑制CD4+T细胞向Th1分化、诱导其凋亡作用,其作用呈剂量依赖关系。
The purpose of the study is to investigate the effect of resveratrol（Res） on the proliferation,apotosis and subsets distribution of murine CD4＋ T lymphocytes.Splenic mononuclear cells（SMC） from specific pathogen free（SPF） Balb/c mice（female,age 9-10 weeks） were cultured with Res（0,20,40,or 80 mmol/L） for 30 min in vitro,then cocultured with Ionomycin and PMA for 1 h for activation,followed by cultivation with protein transporting inhibitor Golgiplug for 4-6 hours.IL-4 and IFN-γ levels in the cultured cells were detected using intracellular cytokine staining with flow cytometry.On the other way,freshly isolated CD4＋T lymphocytes using magnetic cell sorting were cultured with Res（0,10,20,or 40 mmol/L） for 36 hours.Then the cells were stained with PI and Annexin V and assayed using flow cytometry.For detecting the proliferation,CD4＋T lymphocytes were stained with CFSE and cocultured with anti-CD3,anti-CD28 and IL-2 and Res（0,10,20,or 40 mmol/L） for 4 days in vitro.The results showed that Res（≥20 mmol/L） decreased IFN-γ levels and proliferation of CD4＋T lymphocytes significantly in a dose-dependent manner,while Res（≥10 mmol/L） induced CD4＋T lymphocytes apoptosis significantly in a dose-dependent manner.Thus we concluded that Res can inhibit proliferation and Th1 subset differentiation of murine CD4＋ T lymphocytes and induce CD4＋ T lymphocyte apoptosis in a dose-dependent manner.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2013年第9期755-759,共5页
Immunological Journal
基金
广州医学院博士科研项目(2012C83)
