渥曼青霉素通过Chemerin/CMKLR1途径抑制Kupffer细胞NLRP3的活性缓解小鼠非酒精性脂肪肝  

Wortmannin inhibits the activity of NLRP3 in Kupffer cells through the Chemerin/CMKLR1 pathway and relieves nonalcoholic fatty liver in mice

作　　者：吴明兵 张文锋[2] 龚建平[2] 苗春木[2] WU Mingbing;ZHANG Wenfeng;GONG Jianping;MIAO Chunmu(Department of General Surgery,the Second People′s Hospital of Chongqing Liangjiang New Area,Chongqing 401123,China;Department of Hepatobiliary Surgery,the Second Affiliated Hospital of Chongqing Medical University,Chongqing 400010,China)

机构地区：[1]重庆两江新区第二人民医院普外科,401123 [2]重庆医科大学附属第二医院肝胆外科,400010

出　　处：《重庆医学》2018年第17期2279-2284,共6页Chongqing medicine

基　　金：国家自然科学基金资助项目(81701957);重庆卫生和计划生育委员会科技项目(2015-2-126)

摘　　要：目的探讨Chemerin诱导枯否细胞(Kupffer cells,KCs)的炎性反应在小鼠非酒精性脂肪肝病(NAFLD)进展中的作用和机制。方法分别用磷脂酰肌醇3-激酶的抑制剂渥曼青霉素作用于预先用Chemerin体外刺激的KCs 2h,以及由高脂饮食饲养的C57BL/6J小鼠12周。然后,利用酶联免疫吸附试验(ELISA)检测细胞上清液及血清细胞因子水平;反转录酶-聚合酶链式反应(RT-PCR)和蛋白免疫印迹(Western blot)法分别检测体内外KCs中目的分子的mRNA和蛋白表达水平;记录小鼠体质量变化并检测小鼠胰岛素耐受和葡糖糖耐受情况;HE染色结合NAS评分评测小鼠肝脏脂肪变性程度。结果在渥曼青霉素处理的Chemerin-KCs或者NAFLD模型小鼠中:细胞上清液和血清中白细胞介素1β(interleukin 1β,IL-1β)和IL-18的水平明显低于单独用Chemerin处理组或者单独高脂饮食组;Chemerin细胞受体(CMKLR1)和炎症小体3样受体蛋白(NLRP3)的mRNA和蛋白表达明显低于单独用Chemerin处理KCs或者单独高脂饮食的小鼠。此外,在高脂饮食单独饲养的小鼠的体质量变化、肝功能变化、胰岛素抵抗情况、葡糖糖抵抗情况及肝脏脂肪指数变化均比渥曼青霉素处理的NAFLD模型小鼠变化明显。结论渥曼青霉素能通过下调CMKLR1和NLRP3的表达减轻由高脂饮食诱导的肝脏脂肪变和KCs介导的炎性反应。Objective To investigate the role and mechanism of inflammatory response of Kupffer cells induced by Chemerin in the progression of non-alcoholic fatty liver disease （NAFLD） in mice. Methods Wortmannin was used to treated on KCs which pre-treated with Chemerin in vitro for two hours,and treated on C57BL/6J mice which was fed with a high-fat die.Levels of cytokines in supernatant/serum were tested by enzyme-linked immunosorbent assays（ELISA）;mRNA and protein levels of KCs′ Chemokine-like receptor 1 （CMKLR） and nucleotide oligomerization domain （NOD）-like receptor family pyrin domain containing 3 （NLRP3） in vivo and vitro were detected by real time polymerase chain reaction（real time-PCR） and Western blot;changes of mouse weight were recorded;insulin resistance and glucose tolerance were detected;the severity of liver steatosis was evaluated by HE staining combined with NAS score. Results The levels of interleukin 1β （IL-1β） and IL-18 in the KCs and mice treated with wortmannin were significantly lower than the KCs treated with Chemerin only and mice fed with high fat diet only.The mRNA and protein levels of CMKLR1 and inflammasome 3 （NLRP3） were significantly lower in the KCs and mice treated with Wortmannin than the KCs treated with Chemerin only and mice fed with high fat diet only.In addition,changes in mouse weight,hepatic steatosis,liver function,insulin resistance and glucose tolerance were much milder in mice treated with Wortmannin than those mice fed with high fat diet. Conclusion Wortmannin alleviates liver steatosis and inflammation mediated by KCs via down-regulating the expression of CMKLR1 and NLRP3 in high fat diet fed mice.

关 键 词：非酒精性脂肪肝 枯否细胞 渥曼青霉素 炎症小体 CHEMERIN 

分 类 号：R575.5[医药卫生—消化系统]