糖基化终产物对P38MAPK/NF-κB通路诱导的肠道L细胞损伤的影响  

Advanced glycation end products-induced inflammation of L cells via p38MAPK/NF-κB signaling pathway

作　　者：周文君 张振[1] 许宁宁[1] 李双喜[1] 陈宏[1] ZHOU Wen-jun;ZHANG Zhen;XU Ning-ning;LI Shuang-xi;CHEN Hong(Department of Endocrinology, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China)

机构地区：[1]南方医科大学珠江医院内分泌科,广州510282

出　　处：《解放军医学杂志》2018年第5期414-418,共5页Medical Journal of Chinese People's Liberation Army

基　　金：广东省科技计划项目(2014A020212489)~~

摘　　要：目的探讨糖基化终产物(AGEs)在其下游信号通路诱导的肠道L细胞(GLUTag)致炎症损伤中的作用及其机制。方法将肠道L细胞分成6组,即空白对照组、BSA对照组、100μg/ml AGEs干预组、200μg/ml AGEs干预组、300μg/ml AGEs干预组、apocynin(NADPH氧化酶阻断剂)+AGEs(200μg/ml)共培养干预组。各组细胞培养24h后,采用ELISA检测胰高血糖素样肽-1(GLP-1)分泌水平和炎症因子肿瘤坏死因子α(TNF-α)、白细胞介素1(IL-1)、IL-6的表达水平,RT-PCR检测p38MAPK及NF-κB p65 mRNA水平,Western blotting检测p38MAPK、磷酸化p38MAPK(p-p38MAPK)和NF-κB p65蛋白表达水平。结果与对照组相比,AGEs干预肠道L细胞后,AGEs浓度越高,GLP-1分泌水平下降越明显,呈剂量依赖性(P<0.05),同时p38MAPK及NF-κB p65 mRNA水平明显升高,亦呈剂量依赖性(P<0.05),而炎症因子TNF-α、IL-1、IL-6分泌水平也随着AGEs浓度的升高而升高。与AGEs干预组相比,在apocynin干预后,NADPH氧化酶活性受到抑制,GLP-1分泌水平明显升高(P<0.05)。此外,p-p38MAPK和NF-κB p65蛋白表达水平变化与上述mRNA变化趋势一致。结论 AGEs与AGEs受体(RAGE)结合后激活NADPH/p38MAPK/NF-κB信号通路,可能是肠道L细胞早期炎症损伤并最终导致GLP-1分泌减少的作用机制之一。Objective To investigate the effect of Advanced glycation end products（AGEs） on the inflammation of GLUTag cells and explore the possible mechanism. Methods Firstly, GLUTag cells were randomly divided into six groups： control group, BSA group, 100μg/ml AGEs group, 200μg/ml AGEs group, 300μg/ml AGEs group and apocynin（NADPH oxidase blockers）＋AGEs（200μg/ml） group. Then, we detected secretion levels of GLP-1 by using ELISA and mRNA of p38 MAPK and NF-κB p65 by RT-PCR in indicated group. The phospho-p38 MAPK（p-p38 MAPK） and the expression of NF-κB p65 protein were detected by Western blotting. Finally, ELISA was conducted to measure the secretion levels of TNF-α, IL-1, IL6. ResultsCompared with non-treated group（vs NC group, BSA group, P〈0.05）, AGEs treated-group had decreased secretion of GLP-1 which in a dose-dependent manner（P〈0.05）. Meanwhile, AGEs significantly downregulated the mRNA of p38 MAPK and NF-κB p65 in a dose-dependent manner. The secretion of inflammatory cytokines, TNF-α, IL-1, IL-6, were positively related to the dose of AGEs, and AGEs induced-inflammatory effects were abolished by inhibition of NADPH oxidase activity with apocynin, leading to a restoration of GLP-1 secretion. The protein expression of p38 MAPK and NF-κB p65 have the similar trends of change compared with mRNA expression. Conclusion AGEs positively regulate the expression of NADPH oxidase and its down-steam signaling pathway p38 MAPK/NF-κB by targeting RAGES, leading to the inflammation of GLUTag cells. Therefore, we found the pathological activation of p38 MAPK/NF-κB pathway may exert detrimental effect on GLUTag cells and cause inflammation.

关 键 词：糖基化终产物 肠道L细胞 炎症 糖尿病 NF-ΚB通路 

分 类 号：R587.1[医药卫生—内分泌]