中国免疫学杂志
中國免疫學雜誌
중국면역학잡지
Chinese Journal of Immunology
2015年
10期
1315-1319
,共5页
强叶涛%吴水芸%韩慕天%程璐%金慧敏%严成%李翀%刘霞%张苗苗%邵启祥%夏圣
彊葉濤%吳水蕓%韓慕天%程璐%金慧敏%嚴成%李翀%劉霞%張苗苗%邵啟祥%夏聖
강협도%오수예%한모천%정로%금혜민%엄성%리충%류하%장묘묘%소계상%하골
丁酸盐%树突状细胞%共刺激分子%细胞因子%CD8+T细胞%TLR4
丁痠鹽%樹突狀細胞%共刺激分子%細胞因子%CD8+T細胞%TLR4
정산염%수돌상세포%공자격분자%세포인자%CD8+T세포%TLR4
Butyrate%Dendritic cell%Costimulatory molecule%Cytokine%CD8+T%TLR4
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。
目的：觀察腸道細菌代謝產物丁痠對體外培養小鼠骨髓源樹突狀細胞（Bone marrow derived dendritic cells，BMDCs）錶型及功能的影響，併探討其可能的機製。方法：利用丁痠鹽對GM－CSF 和IL－4體外誘導的BMDCs 細胞進行處理，採用流式細胞術檢測BMDCs 細胞錶麵分子CD80、CD86、MHCⅡ、B7－DC 的錶達及其對T 細胞增殖的影響；用實時熒光定量PCR（RT－PCR）檢測其細胞因子IL－6、IL－12的錶達；用格裏斯反應（Griess reaction）和免疫印跡法（Western blot）分彆檢測DC細胞產生NO2－的生成和Toll 樣受體－4（TLR4）信號通路中ERK 分子的燐痠化。結果：丁痠可下調LPS 誘導的成熟BMDCs 細胞CD80、CD86、MHCⅡ、B7－DC 分子的錶達。同時，丁痠也可抑製IL－6和IL－12的分泌，併抑製樹突狀細胞對OVA257－264抗原特異性T 細胞的增殖。進一步的Western blot 檢測結果顯示丁痠可抑製DC 細胞TLR4信號通路中ERK 分子的燐痠化。結論：丁痠可通過抑製DC 細胞TLR4信號通路中ERK 分子的燐痠化，下調DC 細胞的免疫功能。
목적：관찰장도세균대사산물정산대체외배양소서골수원수돌상세포（Bone marrow derived dendritic cells，BMDCs）표형급공능적영향，병탐토기가능적궤제。방법：이용정산염대GM－CSF 화IL－4체외유도적BMDCs 세포진행처리，채용류식세포술검측BMDCs 세포표면분자CD80、CD86、MHCⅡ、B7－DC 적표체급기대T 세포증식적영향；용실시형광정량PCR（RT－PCR）검측기세포인자IL－6、IL－12적표체；용격리사반응（Griess reaction）화면역인적법（Western blot）분별검측DC세포산생NO2－적생성화Toll 양수체－4（TLR4）신호통로중ERK 분자적린산화。결과：정산가하조LPS 유도적성숙BMDCs 세포CD80、CD86、MHCⅡ、B7－DC 분자적표체。동시，정산야가억제IL－6화IL－12적분비，병억제수돌상세포대OVA257－264항원특이성T 세포적증식。진일보적Western blot 검측결과현시정산가억제DC 세포TLR4신호통로중ERK 분자적린산화。결론：정산가통과억제DC 세포TLR4신호통로중ERK 분자적린산화，하조DC 세포적면역공능。
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.
