白藜芦醇通过Sirtuin1激活足细胞Notch1信号  

Resveratrol activates Notch signalling pathway via Sirtuin 1 in podocytes

作　　者：谢可炜[1] 魏凯[1] 顾明颖[1] 向芃[1] 顾乐怡[1,2] 

机构地区：[1]上海交通大学医学院附属仁济医院肾脏科分子细胞(肾病)实验室,200127 [2]新疆维吾尔自治区喀什地区第二人民医院肾病科

出　　处：《中华肾脏病杂志》2015年第4期296-302,共7页Chinese Journal of Nephrology

基　　金：国家自然科学基金（30971363,81270781）;上海市自然科学基金（08ZR1413200）

摘　　要：目的探索白藜芦醇是否影响足细胞中的Notch1信号通路及其可能的机制。方法分别使用RNA干扰（RNAi）技术和盐酸多西环素（doxycycline，Dox）诱导野生型（WT）和可诱导的Sirtuin1（SIRT1）短发夹RNA（shRNA）转基因（CAGGS）足细胞SIRT1表达下降，重组小鼠Notch信号的Delta配体4（Delta—like ligand4，DLL4）诱导激活Notch1通路。实时荧光定量PCR法检测SIRT1、Notch1通路关键γ分泌酶mRNA及Notch1靶基因Hesl、Hey2的表达。Western印迹法检测足细胞内Notch1胞内段结构域（intracellular domain of Notch1，ICD1）、SIRT1、解聚素金属蛋白酶（ADAM）10以及γ分泌酶复合物Presenilin1和Nicastrin的表达。结果白藜芦醇剂量依赖地诱导野生型小鼠足细胞Notch1信号胞内段ICDl表达以及下游产物Hesl和Hey2 mRNA表达增高。白藜芦醇可诱导Notch1蛋白水解酶1分泌酶组分Nicastrin的mRNA和蛋白表达上升以及ADAM10蛋白的表达下降（P〈0．05），而这些变化在使用SIRTIRNAi后均被减弱（P〈0．05）。重组小鼠DLL4可以增加Notch1通路靶基因Hes1和Hey2的mRNA水平，同时增加细胞核内ICD1的表达，盐酸多西环素孵育CAGGS足细胞显著降低SIRT1mRNA水平（P〈0．05），48h后SIRT1蛋白表达明显下降（P〈0．05），并且显著减弱DLIA的作用（P〈0．05）。结论白藜芦醇可以通过高表达分泌酶Nicastrin诱导足细胞中Notch1信号活化，SIRT1介导了白藜芦醇和DLL4诱导Notch1信号活化的作用。Objective To explore the relationship between resveratrol and Notch 1 signalling pathway in podocytes. Methods Interference RNA （RNAi） and doxycycline （Dox） were used to inhibit the Sirtuin （SIRT） 1 expression in the wild- type and inducible SIRT1 shRNA （CAGGS） podocytes respectively. Recombinant mouse delta-like ligand 4 （DLIA） was used to activate Notchl signalling. The message RNA of SIRT1, Notchl downstream gene Hesl and Hey2, as well as the key enzymes of Notchl signalling pathway were detected by using real-time PCR. Western blotting was used to detect intracellular domain of Notch 1 （ICD1）, SIRT1, and metalloprotease （ADAM） 10 and components of γ-secretase complex protein expression. Results In WT murine podoytes, resveratrol up-regnlated ICD1 protein production, as well as the mRNA of Hes1 and Hey2 in a dose-dependent manner. Treatment with resveratrol resulted Nicastrin mRNA and protein increase in podocytes （P 〈 0.05）, as well as inhibit ADAM10 expression （P 〈 0.05）, but all these changes were prevented after the use of SIRT1 RNAi（P 〈 0.05）. DLIA up-regulated the expression of mRNA of Hesl and Hey2, as well as ICD1 protein production in a dose-dependent manner. Treatment with doxycycline resulted decrease of SIRT1 gene and protein expression in CAGGS podocytes after 24 h and 48 h respectively（P 〈 0.05）, which weakend the role of DLIA significantly（P 〈 0.05）. Conclusion Resveratrol induces Nicastrin expression, as well as activation of Notchl signalling pathway in a SIRT1-dependent manner.

关 键 词：足细胞 受体 NOTCHL SIRTUIN 1 Γ-分泌酶 

分 类 号：R692[医药卫生—泌尿科学]