摘要
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 investigate the effect of butyrate produced by bacterial metabolism on immune features of murine bone marrow-derived dendritic cells（BMDCs） and its potential mechanisms.Methods： BMDCs were prepared from bone marrow cells of C57BL/6 mice by being cultured with GM-CSF and IL-4.The expression of CD80,CD86,B7-DC and MHCⅡ on BMDCs and its pri-ming ability on the proliferation of OVA257-264 antigen specific CD8＋T cell were analyzed by using Flow cytometry.The mRNA levels of IL-6 and IL-12 in BMDCs were detected by real-time fluorescence quantitative PCR（q-PCR）.Simultaneously,Griess reaction and Western blot was used for analyzing the levels of NO2-in BMDCs culture supernatant and the ERK phosphortylation in BMDCs respectivly.Results：Butyrate could decrease the levels of CD80,CD86,MHCⅡand B7-DC,and downregulate the capability of BMDCs in priming the proliferation of CD8＋T cells.Furthermore,the secretions of IL-6,IL-12,NO2-and the phosphorylation of ERK were sup-pressed.Conclusion：Butyrate down-regulats the immune functions of BMDCs via inhibition of ERK phosphorylation in TLR 4 signaling pathway.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2015年第10期1315-1319,共5页
Chinese Journal of Immunology
基金
国家自然科学基金(81172834
31428006
31570879)
江苏大学医学临床科技发展基金(JLY20140048)资助
