瘦素对人近端肾小管上皮细胞肝细胞生长因子表达的影响  

Effect of leptin on the hepatocyte growth factor expression of human proximal tubular epithelial cells

作　　者：王沛 仇方忻 姜国珍 姜玲[3] Wang Pei;Qiu Fangxin;Jiang Guozhen;Jiang Ling(Department of Endocrinology,the People's Hospital of Chengyang District,Qingdao 266109,China;Department of Nephrology,the People's Hospital of Chengyang District,Qingdao 266109,China;Department of Blood Purification Center,Qingdao Municipal Hospital,Qingdao 266011,China)

机构地区：[1]青岛市城阳区人民医院内分泌科,青岛266109 [2]青岛市城阳区人民医院肾内科,青岛266109 [3]青岛市市立医院血液净化中心,青岛266011

出　　处：《国际泌尿系统杂志》2020年第4期684-688,共5页International Journal of Urology and Nephrology

基　　金：青岛市城阳区民生引导计划项目(2018-23-09)。

摘　　要：目的探讨瘦素(LEP)诱导的人近端肾小管上皮细胞(HK-2)肝细胞生长因子(HGF)的表达。方法用DMEM/F12培养液培养HK-2,分别用不同浓度的LEP(10、50、100 ng/mL)刺激体外培养的HK-2细胞48 h,以及100 ng/mL的LEP刺激不同的时间(24、48、72 h)。倒置显微镜观察HK-2形态;RT-PCR检测HGF和转化生长因子-β1(TGF-β1)mRNA的表达;免疫细胞化学法检测α-平滑肌肌动蛋白(α-SMA)蛋白表达;ELISA法检测细胞培养液中HGF和TGF-β1蛋白含量。结果①LEP刺激可以使HK-2逐渐由椭圆形变成长梭形,类似肌成纤维细胞的形态;②不同浓度的LEP可以上调HGF和TGF-β1 mRNA及其相应蛋白的表达,其作用有一定的剂量依赖性;③同浓度LEP作用下的HK-2早期即出现HGF mRNA高表达,48 h后表达逐渐下降,之后TGF-β1 mRNA出现表达快速持续上升;细胞培养液中HGF和TGF-β1蛋白表达量与各自相应mRNA表达相似;④α-SMA随LEP浓度的提高或作用时间延长均呈持续性高表达。结论LEP参与了HK-2转分化,它可以在HK-2转分化过程诱导HGF高表达,该表达的上升是一种有益的防御反应。Objective To investigate the expression of hepatocyte growth factor in the process of transdifferentiation of leptin(LEP)-induced human proximal tubular epithelial cell lines.Methods Human renal tubular epithelial cell lines(HK-2)was cultured in DMEM/F12 medium.Cultured HK-2 cells were divided into control group and LEP-treated groups(10,50,100 ng/mL for 48 h,respectively;100 ng/mL for 24,48,72 h,respectively).The cell morphological changes were traced with inverted microscope.The expression of HGF and TGF-β1 at transcriptional were detected by real-time quantitative reverse transcription polymerase chain reaction.The expression ofα-SMA protein was detected by Immunocytochemical.HGF and TGF-β1 secreted into the culture supernatant was assessed by ELISA.Results①LEP stimulated the oval-to-fusiform transdifferentiation in morphology of HK-2 cells.②Compared with control group,both HGF,TGF-β1 mRNA and HGF,TGF-β1 protein was up-regulated by the different concentrations of LEP.③The expression of HGF mRNA increased in an early stage with the effect of leptin,while 48 hours later declined gradually.Then the expression of TGF-β1 mRNA elevated rapidly and continuously.Similar changes of HGF and TGF-β1 expression could also be observed in protein level.④With the concentrations of leptin increased or duration of action,α-SMA showed a persistent high expression.Conclusions LEP involves in HK-2 transdifferentiation.High expression of HGF induced by LEP in HK-2 cell transdifferentiation process,which is a useful defense response.

关 键 词：肾小管 近端 细胞 培养的 瘦素 肝细胞生长因子 转化生长因子Β1 

分 类 号：R692[医药卫生—泌尿科学]