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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。
Objective To study the effects of resveratrol on proliferation, apoptosis and subpopulation of mouse CD4 + T cells. Methods Splenic mononuclear cells (SMCs) of Balb / c mice (SPF, female, 9-10 weeks old) were co-cultured with resveratrol (0,20,40,80 mmol / L) for 30 min. After co-cultured with PMA for 1 h, Golgiplug, a protein transport blocker, was co-cultured for another 4 to 6 h. The cells were harvested for detection of IL-4 and IFN-γ by cytokine staining and flow cytometry. CD4 + T cells were sorted from SMCs by immunomagnetic beads method. CD4 + T cells were co-cultured with resveratrol, anti-CD3 / anti-CD28 antibody and IL-2 at different concentrations (0, 10, 20, 40 mmol / L). After 4 days, the cells were harvested for flow cytometry ; CD4 + T cells incubated with different concentrations of resveratrol (0, 10, 20, 40 mmol / L) for 36 h, the apoptosis was detected by flow cytometry. Results Resveratrol (≥20 mmol / L) could significantly inhibit the proliferation of CD4 + T cells (P <0.05) and inhibit the expression of IFN-γ of CD4 + T lymphocytes (P <0.05), and the inhibitory effect was dose-dependent . Resveratrol (≥10 mmol / L) significantly induced apoptosis of CD4 + T lymphocytes (P <0.01), and the effect was dose-dependent. Conclusion Resveratrol can inhibit the proliferation of CD4 + T cells, inhibit the differentiation of CD4 + T cells to Th1 and induce apoptosis of CD4 + T cells in a dose-dependent manner.