山奈酚对小鼠T淋巴细胞体外活化、增殖和细胞周期的影响

Effects of Kaempferol on activation,proliferation and cell cycle of mouse T lymphocytes in vitro

目的:研究山奈酚(KA)对小鼠T细胞体外活化、增殖和细胞周期的影响,并对其免疫调节作用进行初步探讨。方法:分离小鼠淋巴结细胞,加入多克隆刺激剂刀豆蛋白A(ConA)进行刺激,以不同终浓度的KA与T细胞共培养。流式细胞术结合双色荧光抗体染色,检测T细胞早期活化抗原CD69的表达;MTT法检测KA对T细胞增殖的影响;用碘化丙锭(PI)染色分析T细胞的细胞周期。结果:终浓度为10、20和40μmol/L的KA可以显著抑制ConA刺激的T细胞CD69的表达,由ConA组的(39.11±1.17)%,分别降低为(30.64±0.23)%、(27.95±0.04)%和(5.63±0.37)%(P<0.05);MTT法结果显示,KA对ConA诱导的小鼠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;PI染色结果显示,随着KA浓度的增加,均能明显阻止淋巴细胞进入S期和G2/M期。结论:KA可显著抑制ConA刺激下的小鼠T淋巴细胞体外活化和增殖;它能抑制T淋巴细胞进入细胞分裂期。

AIM： To study the effects of Kaempferol on activation, proliferation and cell cycle of murine T lymphocytes, and to elucidate the mechanism of the immunosuppressive effect of Kaempferol. METHODS： Lymphocytes were prepared from lymphoid nodes of mouse, and were stimulated with polyclonal activators ConA, and then were co-cultured with Kaempferol of different final concentration. The effect on the expression of CD69 （the early marker of the activated T cells） on T lymphocytes were measured by flow cytometry combined with two colored monoclonal antibodies flow cytometry. While, the effect of Kaempferol on the proliferation of T lymphocytes in response to ConA stimulation was determined by MTT, cell cycle was detected by PI staining. RESULTS： Kaempferol （final concentration is 10, 20 and 40 iJmol/L） can inhibit the expression of CD69 on activated T lymphocytes in a dose-dependent manner （ P 〈 0.05 ）. The expression rate of CD69 on T cells in response to ConA was （39.11±1.17）%. After treatment with Kaempferol, the expression rate of CD69 reduced to （30.64 ± 0.23 ） %, （27.95 ± 0.04） % and （5.63 ± 0.37 ） %, respectively （ P 〈 0.05） ; The result of MTT showed that Kaempferol can inhibit the proliferation of T lymphocytes stimulated by ConA in a dose-dependent manner （ P 〈 0.05） ; The result of PI staining showed that Kaempferol can arrest cell cycle at S phage and G2/M at all experimental concentration （ P 〈 0.05）. CONCLUSION： Kaempferol can effectively inhibit the activation, proliferation of mouse T lymphocytes in respond to ConA, arrest cell cycle at S phage and G2/M in vitro.