摘要
目的观察JAK2/STAT3信号途径在IL-6抑制LPS诱导的小鼠髓源性树突状细胞(bone marrow-derived dendritic cells,BMDCs)分化成熟中的作用。方法分离小鼠骨髓细胞,采用黏附法结合GM-CSF和IL-4刺激法在体外培养BMDCs;用相差倒置显微镜观察BMDCs的形态特点;用LPS诱导DCs成熟;采用流式细胞术观察IL-6处理与否,LPS诱导的BMDCs在表型上的变化;Western blot检测BMDCs细胞质p-JAK2、总STAT3和p-STAT3的表达水平。结果形态观察和FCM分析的结果表明:体外成功培养了纯度较高的高表达CD11c的BMDCs,LPS能明显诱导BMDCs表面MHC-Ⅱ类分子和共刺激分子CD80、CD86、CD40的表达上调;而经过IL-6处理后,LPS诱导BMDCs表面MHC-Ⅱ类分子、CD80、CD86、CD40表达上调的能力被显著削弱(P<0.01)。Western blot结果显示,与单独用LPS处理的BMDCs相比,经IL-6处理后的BMDC中p-JAK2、p-STAT3表达随作用时间增加而明显上调,提示IL-6促进JAK2/STAT3信号途径活化。结论JAK2/STAT3信号途径参与了IL-6抑制LPS诱导的BMDCs成熟过程。
Objective To investigate the role of JAK2/STAT3 signaling pathway in IL-6-induced suppression during the maturation of mouse bone marrow-derived dendritic cells （BMDCs） after LPS treatment. Methods To obtain immature BMDCs, mouse bone marrow cells were isolated and cultured in complete culture medium containing GM-CSF and IL-4 after removing the nonadherent cells. The cell morphology was observed by using a phase contrast inverted microscope. LPS was added into the culture medium of BMDCs to induce the cell maturation. The expression of costimulators and MHC class Ⅱ molecules on the surface of LPS- treated BMDCs with or without IL-6 treatment was analyzed by flow cytometry. The expression of p-JAK2, STAT3 and p-STAT3 were detected by Western blot analysis. Results DCs were obtained from mouse bone marrow with normal morphology and high purity. Compared with BMDCs incubated with LPS only, LPS-treated DCs cultured in the presence of IL-6 expressed much lower levels of CD40, CD80, CD86 and MHC class Ⅱ molecules. The expression level of cytoplasmic p-JAK2 and p-STAT3 in BMDCs treated with both LPS and IL-6 were much higher than in BMDCs treated with LPS only. Conclusion JAK2/STAT3 pathway may be involved in the suppression of LPS-induced DCs maturation by IL-6.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2009年第11期1063-1065,共3页
Journal of Third Military Medical University
