【摘要】 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。更多还原

【Abstract】 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-1β and 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-1β or 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-1β and 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-1β and 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.更多还原