STAT3信号通路在IL-6抑制人单核细胞来源树突状细胞CCR7表达中的作用

发布时间：2018-09-01 15:31

【摘要】：目的探讨STAT3信号通路在IL-6抑制人单核细胞来源树突状细胞(human monocytederived dendritic cells,mo DCs)趋化因子受体7(C-C chemokine receptor type 7,CCR7)表达中的作用。方法采用CD14+磁珠分离法从外周血中分离得到单核细胞,加入rh GM-CSF和rh IL-4在体外诱导培养树突状细胞。LPS用于刺激mo DCs表达CCR7,RT-PCR检测不同浓度IL-6处理后再用LPS刺激后的mo DCs CCR7 mRNA表达。Western blot检测IL-6处理mo DCs和IL-6预处理后再加入STAT3磷酸化特各异性抑制剂JSI-124的mo DCs STAT3、p-STAT3表达。IL-6预处理并抑制STAT3磷酸化后,LPS刺激48 h,RT-PCR检测mo DCs CCR7 mRNA的表达,流式细胞仪检测mo DCs CCR7蛋白表达,细胞迁移实验检测mo DCs细胞迁移功能。结果与单独LPS刺激组比较,IL-6明显抑制LPS刺激的mo DCs CCR7mRNA的表达(P0.05)。Western blot结果显示,与单独LPS刺激组比较,IL-6导致mo DCs细胞内STAT3高度活化;而抑制STAT3信号高度活化后,与IL-6+LPS处理组比较,mo DCs CCR7 mRNA表达、蛋白分子表达、细胞迁移能力明显升高(P0.05),与LPS刺激组无差异(P0.05)。结论 IL-6通过高度活化STAT3信号通路抑制人单核细胞来源树突状细胞CCR7表达,导致树突状细胞的迁移能力受损,而靶向干预STAT3通路可能成为纠正树突状细胞迁移障碍的潜在手段。
[Abstract]:Objective to investigate the role of STAT3 signaling pathway in inhibiting the expression of chemokine receptor 7 (C-C chemokine receptor type 7 CCR7) in human monocyte derived dendritic cells (human monocytederived dendritic cells,mo DCs) by IL-6. Methods Mononuclear cells were isolated from peripheral blood by CD14 magnetic beads method. Induction of dendritic cells with rh GM-CSF and rh IL-4 in vitro. LPs was used to stimulate mo DCs expression CCR7,RT-PCR to detect mo DCs CCR7 mRNA expression after IL-6 treatment with different concentrations and mo DCs CCR7 mRNA expression after LPS stimulation. Western blot was used to detect mo DCs and IL-6 pretreatment with IL-6 and IL-6. The mo DCs STAT3,p-STAT3 expression of JSI-124, a special inhibitor of STAT3 phosphorylation, was pretreated with IL-6 and the expression of mo DCs CCR7 mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR) at 48 h after STAT3 phosphorylation. The expression of mo DCs CCR7 protein was detected by flow cytometry, and the migration function of mo DCs cells was detected by cell migration assay. Results compared with LPS alone, IL-6 significantly inhibited the expression of mo DCs CCR7mRNA stimulated by LPS (P0.05). Western blot results showed that IL-6 induced STAT3 hyperactivation in mo DCs cells compared with that in LPS alone, but inhibited the high activation of STAT3 signal. Compared with IL-6 LPS treatment group, the expression of DCs CCR7 mRNA, protein molecule expression and cell migration ability were significantly increased (P0.05), but no difference was found between LPS group and LPS group (P0.05). Conclusion IL-6 inhibits the expression of CCR7 in dendritic cells derived from human monocytes through highly activated STAT3 signaling pathway, resulting in impaired migration ability of dendritic cells. Targeted intervention of STAT3 pathway may be a potential means to correct the dysfunction of dendritic cell migration.
【作者单位】：第三军医大学西南医院全军普通外科中心微创胃肠外科中心;
【基金】：国家自然科学基金面上项目(81071980,81372560)~~
【分类号】：R966

【参考文献】