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芹菜素对小鼠脾细胞中树突状细胞成熟及功能的影响

[Effect of apigenin on dendritic cells maturation and function in murine splenocytes].

作者信息

Liu Yi-fei, Xue Xiao-xu, Li Zheng-yi, Wang Jun-peng, Zhang Yi-jie

出版信息

Yao Xue Xue Bao. 2017 Mar;52(3):397-402.

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.

摘要

本研究旨在探讨芹菜素(Api)对小鼠脾细胞中树突状细胞(DCs)成熟及功能的影响。分离单个脾细胞,然后在有或无芹菜素的情况下与脂多糖(LPS)共同培养。24小时后,使用CCK8试剂盒测定Api的毒性和T细胞增殖情况。此外,我们收集无细胞上清液,采用酶联免疫吸附测定法(ELISA)检测细胞因子的产生,收集细胞,使用流式细胞术测定DC成熟情况。最后,我们纯化经Api和/或LPS处理的CD11c+DCs,用卵清蛋白(OVA)323−339脉冲处理后,过继转移至C57BL/6小鼠体内,分别检测OVA323−339特异性T细胞增殖以及辅助性T细胞(Th1)和Th2细胞分泌IFN-γ和IL-4的情况。我们发现,Api不影响脾细胞活力,但抑制促炎细胞因子IL-1β、IL-6和TNF-α的产生,而不影响抗炎细胞因子IL-10的产生。此外,Api抑制CD11c+DCs共刺激分子CD80、CD86和MHCII的表达。最后,与LPS+OVA DCs组相比,来自Api和LPS共同处理的脾细胞的DCs(Api+LPS+DCs)损害了OVA323−339特异性T细胞增殖以及CD4+T细胞中IFN-γ和IL-4的产生,其反应与OVA+DCs相似。这些数据表明,Api通过抑制DC激活和功能发挥抗炎特性,作为一种新的免疫调节剂,可能诱导免疫耐受,对慢性炎症患者有益。

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