论文部分内容阅读
目的:观察肠道细菌代谢产物丁酸对体外培养小鼠骨髓源树突状细胞(Bone marrow derived dendritic cells,BMDCs)表型及功能的影响,并探讨其可能的机制。方法:利用丁酸盐对GM-CSF和IL-4体外诱导的BMDCs细胞进行处理,采用流式细胞术检测BMDCs细胞表面分子CD80、CD86、MHCⅡ、B7-DC的表达及其对T细胞增殖的影响;用实时荧光定量PCR(RT-PCR)检测其细胞因子IL-6、IL-12的表达;用格里斯反应(Griess reaction)和免疫印迹法(Western blot)分别检测DC细胞产生NO2-的生成和Toll样受体-4(TLR4)信号通路中ERK分子的磷酸化。结果:丁酸可下调LPS诱导的成熟BMDCs细胞CD80、CD86、MHCⅡ、B7-DC分子的表达。同时,丁酸也可抑制IL-6和IL-12的分泌,并抑制树突状细胞对OVA257-264抗原特异性T细胞的增殖。进一步的Western blot检测结果显示丁酸可抑制DC细胞TLR4信号通路中ERK分子的磷酸化。结论:丁酸可通过抑制DC细胞TLR4信号通路中ERK分子的磷酸化,下调DC细胞的免疫功能。
OBJECTIVE: To observe the effect of intestinal bacterial metabolite butyric acid on the phenotype and function of murine bone marrow-derived dendritic cells (BMDCs) in vitro and to explore its possible mechanism. Methods: The cells of BMDCs induced by GM-CSF and IL-4 were treated by butyrate. The expression of CD80, CD86, MHCⅡ and B7-DC on BMDCs were detected by flow cytometry and their effects on the proliferation of T cells The expression of cytokines IL-6 and IL-12 were detected by real-time fluorescence quantitative PCR (RT-PCR). The expression of NO2- was detected by Griess reaction and Western blot respectively Generated and phosphorylated ERK molecules in the Toll-like Receptor-4 (TLR4) signaling pathway. Results: Butyric acid could down-regulate the expression of CD80, CD86, MHCⅡ and B7-DC in LPS-induced BMDCs. Butyric acid also inhibits the secretion of IL-6 and IL-12 and inhibits the proliferation of dendritic cells against OVA257-264 antigen-specific T cells. Further Western blot results showed that butyric acid could inhibit the phosphorylation of ERK in TLR4 signaling pathway in DCs. CONCLUSION: Butyric acid can downregulate the immune function of DC cells by inhibiting the phosphorylation of ERK in the TLR4 signaling pathway of DCs.