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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。
Objective: To study the effect of kaempferol (KA) on the activation, proliferation and cell cycle of murine T cells in vitro and to explore its immunoregulatory effect. Methods: Mouse lymph node cells were isolated and stimulated with polyclonal antibody ConA. KA and T cells were co-cultured with different concentrations of KA. The expression of CD69, an early activation antigen of T cells, was detected by flow cytometry and double color fluorescent antibody staining. The effect of KA on the proliferation of T cells was detected by MTT assay. The cell cycle of T cells was analyzed by propidium iodide (PI) staining. Results: KA at final concentrations of 10, 20 and 40 μmol / L significantly decreased the expression of CD69 in ConA-stimulated T cells by (39.11 ± 1.17)% and (30.64 ± 0.23)% and (27.95 ± 0.04)% and (5.63 ± 0.37)%, respectively (P <0.05). The results of MTT assay showed that KA had a significant inhibitory effect on T lymphocytes proliferation induced by ConA (P <0.05) PI staining showed that with the increase of KA concentration, lymphocytes could be prevented from entering into S phase and G2 / M phase. Conclusion: KA can significantly inhibit the activation and proliferation of mouse T lymphocytes stimulated by ConA; it can inhibit T lymphocytes into the cell division.