胰高血糖素样肽-1通过下调二肽基肽酶-4/核因子-κB信号通路抑制氧化应激诱导小鼠胚胎成纤维细胞炎症的机制  

Glucagon like peptide-1 down-regulates dipeptidyl peptidase-4/nuclear factor-κB signaling pathway to inhibit mouse embryonic fibroblasts inflammation induced by oxidative stress

作　　者：买买提·依斯热依力 仙米西努尔·买买提明 依力汗·依明 王永康 阿巴伯克力·乌斯曼 克力木·阿不都热依木[2] Maimaiti·Yisireyili;Xianmixinuer·Maimaitiming;Yilihan·Yiming;Wang Yongkang;Ababokeli·Wusiman;Kelimu·Abudureyimu(Research and Education Center,People’s Hospital of Xinjiang Uygur Autonomous Region,Urumqi 830001,China;Research Institute of General and Minimally Invasive Surgery of People’s Hospital of Xinjiang Uygur Autonomous Region,Urumqi 830001,China)

机构地区：[1]新疆维吾尔自治区人民医院科研教育中心,乌鲁木齐830001 [2]新疆维吾尔自治区人民医院自治区普外微创研究所,乌鲁木齐830001

出　　处：《中华实验外科杂志》2024年第5期951-955,共5页Chinese Journal of Experimental Surgery

基　　金：新疆维吾尔自治区引进高层次人才天池百人计划项目(201939);国家自然科学基金(82060166)。

摘　　要：目的:探讨胰高血糖素样肽-1(GLP-1)通过调控NADPH氧化酶-4(Nox-4)及二肽基肽酶-4(DPP-4)/核因子-κB(NF-κB)信号通路,影响脂肪炎症的发生及其相关机制。方法:小鼠胚胎成纤维细胞(3T3-L1细胞,Procell,CL-0006)进行传代培养并进一步分为空白对照组(Control)、加入5μg/ml内质网应激诱导剂衣霉素(TM)组及GLP-1组(加入10 nmol/L GLP-1预处理24 h后,加入TM),3组培养24 h;在3T3-L1脂肪细胞中用小干扰RNA(siRNA)沉默DPP-4的表达,后分别提取总RNA和提取蛋白,其浓度测定并进行实时反转录-聚合酶链反应(RT-PCR)和蛋白质印迹法(Western blot)实验分析Nox-4、GLP-1受体(GLP-1R)、NF-κB亚基(p-50、p-65)蛋白以及炎性因子如单核细胞趋化蛋白-1(MCP-1)、白细胞介素-6(IL-6)及肿瘤坏死因子-α(TNF-α)的mRNA和蛋白表达水平,组间均数比较采用方差分析。结果:3T3-L1脂肪细胞加入GLP-1后,GLP-1组GLP-1R的mRNA表达水平高于TM组(1.10±0.05比0.47±0.07),差异有统计学意义(F=183.912,P<0.001);其DPP-4和Nox-4的mRNA表达水平低于TM组(2.68±0.30比1.31±0.15;2.04±0.21比1.38±0.11),差异有统计学意义(F=90.196、60.166,P<0.001)。GLP-1组p-50和p-65的蛋白表达水平低于TM组(1.48±0.23比1.22±0.03;1.66±0.13比1.15±0.06),差异有统计学意义(F=13.049、73.488,P<0.001);GLP-1组MCP-1、IL-6及TNF-α的mRNA相对表达水平低于TM组(1.62±0.18比0.95±0.03;1.80±0.32比1.23±0.02;2.05±0.31比1.30±0.03),差异有统计学意义(F=45.140、19.524,F=35.933,P<0.001)。si-DPP-4敲低组的p-50和p-65的蛋白表达水平低于TM组(1.42±0.07比1.13±0.02;1.59±0.18比1.12±0.03),差异有统计学意义(F=93.111、30.849,P<0.001);si-DPP-4敲低组的Nox-4(2.13±0.10比1.34±0.05,F=219.708,P<0.001)和炎性因子MCP-1、IL-6及TNF-α的mRNA表达水平低于TM组(1.46±0.06比1.19±0.04,1.79±0.13比1.23±0.05,2.26±0.14比1.32±0.07),差异有统计学意义(F=87.228、72.369、153.372,P<0.001)。结论:GLP-1通过下调DPP4/NF-κB信号通路抑�Objective We investigated the mechanisms of how glucagon-like peptide-1(GLP-1)affects adipose inflammation through the regulation of NADPH oxidase-4(Nox-4)and dipeptidyl peptidase-4(DPP-4)/nuclear factor-κB(NF-κB)signaling pathways.Methods Mouse embryonic fibroblasts(3T3-L1 cells,Procell,CL-0006)were cultured and further divided into a control group,TM group(added 5μg/ml of the endoplasmic reticulum stress inducer tunicamycin,TM),and GLP-1 group(cells pre-treated with 10 nmol/L GLP-1 for 24 h,then added TM),these three group cultured for 24 h.The expression of DPP-4 was silenced by small interfering RNA(siRNA)in 3T3-L1 adipocytes.All groups total RNA and protein samples were extracted,and their concentrations were determined.Nox-4,GLP-1 receptor(GLP-1R),NF-κB subunit(p-50,p-65)proteins as well as mRNA and protein expression levels of inflammatory factors such as monocyte chemotactic protein-1(MCP-1),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)analyzed by real-time reverse transcription-polymerase chain reaction(RT-PCR)and Western blotting experiments respectively.The comparison of means between groups was performed by ANOVA.Results After the addition of GLP-1 to 3T3-L1 adipocytes,the mRNA expression level of GLP-1R in the GLP-1 group was higher than that in the TM group(1.10±0.05 vs.0.47±0.07),had a significant difference(F=183.912,P<0.01);and the mRNA expression levels of DPP-4 and Nox-4 were lower than those in the TM group(2.68±0.30 vs.1.31±0.0.15;2.04±0.0.21 vs.1.38±0.11),had a significant difference(F=90.196,60.166,P<0.01).p-50 and p-65 in the GLP-1 group were lower than those in the TM group(1.48±0.23 vs.1.22±0.03;1.66±0.13 vs.1.15±0.06),had a significant difference(F=13.049,73.488,P<0.01);The mRNA relative expression levels of MCP-1,IL-6 and TNF-αin the GLP-1 group were lower than those in the TM group(1.62±0.18 vs.0.95±0.0.03;1.80±0.32 vs.1.23±0.02;2.05±0.02;1.80±0.32 vs.0.02;2.05±0.31 vs.1.30±0.03),had a significant difference(F=45.140,19.524,35.933,P<0.01).siDPP-4 knoc

关 键 词：脂肪细胞 氧化应激 二肽基肽酶-4/核因子-κB信号通路 炎性因子 

分 类 号：R364.5[医药卫生—病理学]