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芹菜素对脾细胞中树突状细胞的成熟和功能的影响
Effect of apigenin on dendritic cells maturation and function in murine splenocytes
【摘要】 探讨芹菜素对小鼠中抗原递呈细胞特别是树突状细胞(dendritic cells,DCs)成熟和递呈抗原能力的影响。体外分离成年小鼠脾细胞后,经脂多糖(lipopolysaccharide,LPS)和芹菜素共培养24 h后,Cell Counting Kit(CCK)-8、酶联免疫吸附实验(enzyme-linked immunosorbent assay,ELISA)和流式细胞术等方法分别检测芹菜素对脾细胞的毒性作用、分泌炎症细胞因子的作用、共刺激分子表达的影响;分选芹菜素和/或LPS处理后的DCs,与卵清白蛋白(ovalbumin,OVA)323-339共孵育并过继转移到正常小鼠体内7天后,分离T细胞并经OVA323-339重新刺激后,CCK-8法和流式细胞仪检测OVA323-339特异性T细胞的增殖和分泌炎症细胞因子的能力。结果显示,芹菜素抑制LPS刺激的脾细胞分泌促炎症细胞因子白细胞介素(interleukin,IL)-1β、IL-6和肿瘤坏死因子(tumor necrosis factor,TNF)-α的产生,但不影响抗炎症细胞因子IL-10的产生,而该效应并非依赖其毒性杀伤作用。进一步研究发现,芹菜素抑制脾细胞CD11c+DCs的共刺激分子CD80、CD86和MHCII的表达,提示芹菜素可能诱导DCs的不成熟或减弱其抗原递呈能力。体内功能实验结果显示,芹菜素处理的DCs刺激抗原特异性T细胞的增殖及分泌细胞因子[辅助性T细胞(Th)1:IFN-γ和Th2:IL-4]的能力显著减弱。综上所述,芹菜素的抗炎作用可能是通过调控脾细胞中DCs的成熟和递呈抗原的能力,从而在一定程度上抑制炎症细胞因子的产生。
【Abstract】 This study was designed to explore the effect of apigenin(Api) on dendritic cell(DCs) maturation and function in murine spleen cells. The single spleen cell was isolated, and then cultured with lipopolysaccharide(LPS) in the present and absence of apigenin. After 24 h, the toxicity of Api and the T cell proliferation were determined by CCK8 kit. In addition, we collected the cell-free supernatants to measure cytokine production using ELISA, collected the cells to determine the DC maturation using flow cytometry. Finally, we purified Api and/or LPS-treated CD11c+ DCs which were pulsed with ovalbumin(OVA)323-339 and then were adoptive transferred into C57BL/6 mice to detect the OVA323-339-specific T cell proliferation and T helper(Th1) and Th2 cell secreting IFN-γ and IL-4 production, respectively. We found that Api did not affect splenocyte viability, but inhibited the production of pro-inflammatory cytokine IL-1β, IL-6 and TNF-α, not anti-inflammatory cytokine IL-10. In addition, Api inhibited the expression of co-stimulatory CD80, CD86 and MHCII of CD11c+ DCs. Finally, compared to LPS+OVA DCs group, DCs from Api and LPS co-treated splenocytes(Api+LPS+DCs) impaired OVA323-339-specific T cell proliferation and the production of IFN-γ and IL-4 in CD4+ T cells, which had the similar responses with OVA+DCs. These data suggest that Api exhibits anti-inflammatory properties via inhibiting DC activation and function, as a new immune-modulator, which may induce immune-tolerance with a benefit to those with chronic inflammation.
【Key words】 apigenin; dendritic cell; antigen-presenting cell; cytokine; immune tolerance;
- 【文献出处】 药学学报 ,Acta Pharmaceutica Sinica , 编辑部邮箱 ,2017年03期
- 【分类号】R392
- 【被引频次】6
- 【下载频次】208