高糖对大鼠肾间质成纤维细胞PINCH-1表达的影响  

Effects of high glucose on expression of PINCH-1 inrenal interstitial fibroblasts from rats

作　　者：谢荣[1,2] 张和平 刘晓惠[1] 刘佳丽[1] 李沁芸[1] XIE Rong;ZHANG He-ping;LIU Xiao-hui;LIU Jia-li;LI Qin-yun(Department of Nephrology,Affiliated Hospital of North Sichuan Medical College,Nanchong 637000,China)

机构地区：[1]川北医学院附属医院肾内科,南充637000 [2]川北医学院,南充637100

出　　处：《临床肾脏病杂志》2020年第4期316-321,共6页Journal Of Clinical Nephrology

基　　金：2019年四川省卫生健康委员会科研课题重点研究项目(19ZD006)。

摘　　要：目的研究高糖对大鼠肾间质成纤维细胞(NRK49F)表达PINCH-1(Particulary interesting new cysteine-histicline rich protein-1)和整合素连接激酶(Integrin-linked kinase,ILK)的影响,探讨PINCH-1在糖尿病肾病中的作用机制。方法(1)体外培养大鼠NRK49F细胞,分别采用高糖(25 mmol/L)和低糖(5.5 mmol/L)培养基培养6 h、12 h、24 h、48 h、72 h后,检测各组细胞不同时间点PINCH-1、ILK、α-平滑肌肌动蛋白(α-Smooth Muscle Actin,α-SMA)mRNA及蛋白表达的动态变化,ELISA法检测各组细胞上清液纤维连接蛋白(fibronectin,FN)、Ⅰ型胶原(collagen type I,Col I)的表达。(2)采用不同浓度葡萄糖(5.5 mmol/L、15.0 mmol/L、25.0 mmol/L)作用于大鼠NRK49F细胞48 h后,检测各组细胞PINCH-1、ILK、α-SMA mRNA及蛋白的表达。结果(1)与低糖组相比,高糖组PINCH-1、ILK、α-SMA mRNA及蛋白的表达明显上调,高糖刺激早期呈一定时间依赖性增加,FN、Col I蛋白的分泌水平随着高糖刺激时间的延长,呈持续升高,差异均有统计学意义(P<0.05)。(2)不同浓度葡萄糖培养大鼠NRK49F细胞48 h后,PINCH-1、ILK、α-SMA mRNA及蛋白的表达呈浓度依赖性增加,FN、Col I的分泌水平随着葡萄糖浓度的增加持续升高。结论高糖能够促进NRK49F中调节蛋白PINCH-1和ILK的表达,促进肾脏成纤维细胞表型转化以及纤维化蛋白FN、Col I的分泌。PINCH-1可能通过与ILK相互作用,在糖尿病肾病肾间质纤维化的发生发展中发挥重要的作用。Objective To observe the effects of high glucose on expression of particulary interesting new cysteine-histicline rich protein-1(PINCH-1)and integrin-linked kinase(ILK)in renal interstitial fibroblasts(NRK49F)from rats,and to investigate the role and possible mechanism of PINCH-1 in the pathogenesis of diabetic kidney disease.Methods(1)NRK49F cells from rats were cultured in vitro in high glucose medium(25 mmol/L)and low glucose medium 5.5 mmol/L ,respectively.After 6 hours,12 hours,24 hours,48 hours and 72 hours,the changes of the mRNA and protein expression of PINCH-1,ILK andα-smooth muscle actin(α-SMA)in these groups were detected by real-time quantitative polymerase chain reaction(RT-qPCR)and immunocytochemistry(ICC).The expressions of fibronectin(FN)and collagen type I(Col I)were determined with ELISA.(2)The NRK49F cells were cultured in the media at different concentrations of glucose(5.5 mmol/L,15.0 mmol/L,25.0 mmol/L).After 48 hours,the mRNA and protein expression of PINCH-1,ILK andα-SMA were detected.Results(1)Compared with the low glucose group,the mRNA and protein expressions of PINCH-1,ILK andα-SMA in the high glucose group were up-regulated significantly,and exhibited a time-dependent at the early stage of high glucose stimulation.The secretion levels of Col I and FN continued to increase with the prolongation of high glucose stimulation.All the differences were of statistically significance(P<0.05).(2)After NRK49F cells were cultured in the media at different concentrations of glucose(5.5 mmol/L,15.0 mmol/L,25.0 mmol/L)for 48 hours,the mRNA and protein expressions of PINCH-1 and ILK increased in a concentration-dependent manner.The secretion levels of Col I and FN continued to increase with the increase of glucose concentration.Conclusions High glucose can promote the expression of the regulatory proteins PINCH-1 and ILK in NPK49F,the phenotype transformation of fibroblasts,and the secretion of fibrotic proteins Col I and FN.PINCH-1 may play an important role in the pathogenesis and progression

关 键 词：PINCH-1蛋白 ILK 高糖 糖尿病肾病 成纤维细胞 

分 类 号：R587.2[医药卫生—内分泌] R692.9[医药卫生—内科学]