糖化白蛋白调控肾损伤相关分子与Toll样受体信号通路及齐墩果酸对其干预作用的研究  

Glycated albumin regulates renal injury-associated molecules and Toll-like receptor signaling pathway and the intervention effect of oleanolic acid

作　　者：段楠[1] 黄辰炜 逄璐 李海霞[1] Duan Nan;Huang Chenwei;Pang Lu;Li Haixia(Department of Clinical Laboratory,Peking University First Hospital,Beijing 100034,China)

机构地区：[1]北京大学第一医院检验科

出　　处：《首都医科大学学报》2019年第4期609-614,共6页Journal of Capital Medical University

基　　金：北京大学第一医院科研种子基金(2019SF35)~~

摘　　要：目的探讨人近端肾小管上皮细胞(HK-2细胞系)在糖化白蛋白(glycated albumin,GA)与齐墩果酸(oleanolic acid,OA)作用下,肾损伤分子-1 (kidney injury molecule-1,KIM-1)、中性粒细胞明胶酶相关载脂蛋白(neutrophil gelatinase-associated lipocalin,NGAL)及Toll样受体信号通路相关分子的表达情况。方法采用GA及OA处理HK-2细胞24 h,real-time PCR检测KIM-1、NGAL以及p38丝裂原活化蛋白激酶(mitogen activated protein kinase,MAPK)、白细胞介素-1受体相关激酶4(interleukin-1receptor-associated kinase 4,IRAK4)、Toll样受体(Toll-like receptor,TLR) 1、TLR2、TLR7和TLR9的mRNA表达,ELISA法检测细胞上清液KIM-1与NGAL水平,Western blotting法检测细胞内p38 MAPK、IRAK4、TLR1、TLR2、TLR7和TLR9蛋白表达。结果与对照组比较,GA能显著上调KIM-1、NGAL mRNA与蛋白的含量(P<0.05),并促进p38 MAPK、IRAK4、TLR1、TLR2 mRNA及蛋白的表达(P<0.05);加入OA能显著抑制上述作用(P<0.05)。结论 GA能够上调KIM-1和NGAL的表达与释放,激活Toll样受体信号通路,对人近端肾小管上皮细胞造成损伤;而加入OA能够有效干预GA调控的损伤。Objective To investigate the effects of glycated albumin(GA) and oleanolic acid(OA) on the expression of kidney injury molecule-1(KIM-1),neutrophil gelatinase-associated lipocalin(NGAL) and some molecules of Toll-like receptor(TLR)signaling pathway in human proximal tubular epithelial cells(HK-2 cells).Methods The HK-2 cells were treated with GA,GA and OA for 24 h.The mRNA levels of KIM-1,NGAL,p38 mitogen activated protein kinase(MAPK),interleukin-1 receptor-associated kinase 4(IRAK4),TLR 1,TLR2,TLR7 and TLR9 were detected by real-time PCR.The releases of KIM-1 and NGAL in the supernatants were detected by ELISA.The protein levels of p38 MAPK,IRAK4,TLR1,TLR2,TLR7 and TLR9 were measured by Western blotting.Results Compared with control group,GA could significantly up-regulate the mRNA and the protein levels released in supernatants of KIM-1 and NGAL(P<0.05),and promoted the expression of p38 MAPK,IRAK4,TLR1,TLR2 mRNA and protein in HK-2 cells(P< 0.05).OA treatment could significantly inhibit theses effects(P<0.05).Conclusion GA can up-regulate the expression and release of KIM-1 and NGAL,and activate the TLR signaling pathway,suggesting that GA could play an important role in regulating renal injury.OA had effective intervention effect on GA.

关 键 词：糖化白蛋白 齐墩果酸 TOLL样受体信号通路 肾损伤分子-1 中性粒细胞明胶酶相关载脂蛋白 

分 类 号：R692.6[医药卫生—泌尿科学]