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作 者:慕静静[1] 曾耀英[1] 黄秀艳[1] 赵晓慧[1] 宋兵[1]
机构地区:[1]暨南大学生命科学技术学院组织移植与免疫中心,广东广州510632
出 处:《细胞与分子免疫学杂志》2009年第12期1106-1108,1111,共4页Chinese Journal of Cellular and Molecular Immunology
基 金:国家重点基础研究发展计划(973)资助项目(2006CB504200;2004CB720100)
摘 要: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.
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