瘦素通过NLRP3 炎性体对小鼠骨髓树突状细胞的作用及机制

目的:探讨瘦素通过NLRP3 炎性体对树突状细胞的作用及其机制。方法:体外诱导小鼠骨髓来源的树突状细胞(BMDCs),用不同浓度瘦素与BMDCs 共同培养,分别检测培养上清IL-1β和IL-18 的蛋白和基因水平,用FLICA 试剂盒或ROS 试剂盒分别在流式细胞仪上检测caspase-1 活性或胞内ROS 生成情况。瘦素与BMDCs 共培养,加入caspase-1 抑制剂或用siRNA 干扰NLRP3 基因表达,检测前后加入IL-1β和IL-18 基因和蛋白表达水平。瘦素与BMDCs 共培养,加入ROS 抑制剂或KCL,检测加入前后IL-1β和IL-18 蛋白分泌水平。结果:瘦素促进BMDCs 分泌IL-1β和IL-18。瘦素通过上调NLRP3 基因促进IL-1β和IL-18 基因和蛋白表达,通过激活caspase-1 蛋白提高IL-1β基因和蛋白水平,K+ 外流参与此过程。结论:瘦素通过激活NLRP3 炎性体促进BMDCs IL-1β和IL-18 基因和蛋白表达,此过程部分通过K+ 外流实现。这提示瘦素可能是NLRP3 炎性体的激活剂。

Impact and mechanism of leptin on bone marrow-derived dendritic cells(BMDCs)via NLRP3 inflammasome

Objective:To investigate the role and mechanism of leptin on dendritic cells by NLRP3 inflammasome.Methods: BMDCs were induced in vitro,leptin with scalar doses was cocultured with BMDCs,IL-1βand IL-18 mRNA expression and protein secretion level were measured by q-RT-PCR and ELISA respectively.Caspase-1 activity or ROS synthesis were tested with FLICA kit or ROS detection assay kit on flow cytometry.IL-1βor IL-18 were detected after caspase-1 was inhibited by Ac-YVAD-cmk or NLRP3 was interfered by siRNA or ROS inhibitor DPI or KCL were added.Results: Leptin promoted secretion of IL-1βand IL-18.Leptin up-regulated NLRP3 and activted caspase-1 to secret proinflammtory cytokine,which K+ efflux took part in.Conclusion: Leptin promotes secretion of IL-1βand IL-18 by activating NLRP3 inflammasome,and K+ efflux takes part in this,which hints us that leptin may be an activator of NLRP3 inflammasome.