Globular adiponectin-mediated vascular remodeling by affecting the secretion of adventitial-derived tumor necrosis factor-αinduced by urotensinⅡ  

作　　者：Jun LI Limin LUO Yonggang ZHANG Xiao DONG Shuyi DANG Xiaogang GUO Wenhui DING 

机构地区：[1]Department of Cardiology,the First Affiliated Hospital,Zhejiang University School of Medicine,Hangzhou 310003,China [2]Division of Cardiology,Department of Internal Medicine,Taihe Hospital,Hubei University of Medicine,Shiyan 442000,China [3]Division of Cardiology,Department of Internal Medicine,Peking University First Hospital,Bejing 100034,China [4]Department of Dermatology,the First Affiliated Hospital,Zhejiang University School of Medicine,Hangzhou 310003,China [5]Department of Cardiovascular Diseases,the Second Affiliated Hospital,Shantou University Medical College,Shantou 515041,China

出　　处：《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》2022年第12期1014-1027,共14页浙江大学学报（英文版）B辑（生物医学与生物技术）

基　　金：supported by the the National Natural Science Foundation of China(No.82003372);the Natural Science Foundation of Hubei Province(Nos.2018CFB747 and 2018CFB537);the Educational Commission of Hubei Province(Nos.B2017112 and B20181130),China.

摘　　要：Objectives:In this study,we explored how adiponectin mediated urotensinⅡ(UⅡ)-induced tumor necrosis factor-α(TNF-α)andα-smooth muscle actin(α-SMA)expression and ensuing intracellular signaling pathways in adventitial fibroblasts(AFs).Methods:Growth-arrested AFs and rat tunica adventitia of vessels were incubated with UⅡand inhibitors of signal transduction pathways for 1-24 h.The cells were then harvested for TNF-αreceptor(TNF-α-R)messenger RNA(mRNA)and TNF-αprotein expression determination by reverse transcription-polymerase chain reaction(RT-PCR)and enzyme-linked immunosorbent assay(ELISA),respectively.Adiponectin and adiponectin receptor(adipoR)expression was measured by RT-PCR,quantitative real-time PCR(qPCR),immunohistochemical analysis,and cell counting kit-8(CCK-8)cell proliferation experiments.We then quantified TNF-αandα-SMA mRNA and protein expression levels by qPCR and immunofluorescence(IF)staining.RNA interference(RNAi)was used to explore the function of the adipoR genes.To investigate the signaling pathway,we applied western blotting(WB)to examine phosphorylation of adenosine 5’-monophosphate(AMP)-activated protein kinase(AMPK).In vivo,an adiponectin(APN)-knockout(APN-KO)mouse model mimicking adventitial inflammation was generated to measure TNF-αandα-SMA expression by application of qPCR and IF,with the goal of gaining a comprehensive atlas of adiponectin in vascular remodeling.Results:In both cells and tissues,UⅡpromoted TNF-αprotein and TNF-α-R secretion in a dose-and time-dependent manner via Rho/protein kinase C(PKC)pathway.We detected marked expression of adipoR1,T-cadherin,and calreticulin as well as a moderate presence of adipoR2 in AFs,while no adiponectin was observed.Globular adiponectin(gAd)fostered the growth of AFs,and acted in concert with UⅡto induceα-SMA and TNF-αthrough the adipoR1/T-cadherin/calreticulin/AMPK pathway.In AFs,gAd and UⅡsynergistically induced AMPK phosphorylation.In the adventitial inflammation model,APN deficiency up-regulated the expre

关 键 词：Urotensin II ADIPONECTIN Signal transduction Adventitial fibroblast RNA interference(RNAi) Adiponectin-knockout(APN-KO) 

分 类 号：R54[医药卫生—心血管疾病]