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

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2022年第12期23卷 1014-1027页

作者： Jun LI Limin LUO Yonggang ZHANG Xiao DONG Shuyi DANG Xiaogang GUO Wenhui DING Department of Cardiology the First Affiliated HospitalZhejiang University School of MedicineHangzhou 310003China Division of Cardiology Department of Internal MedicineTaihe HospitalHubei University of MedicineShiyan 442000China Division of Cardiology Department of Internal MedicinePeking University First HospitalBejing 100034China Department of Dermatology the First Affiliated HospitalZhejiang University School of MedicineHangzhou 310003China Department of Cardiovascular Diseases the Second Affiliated HospitalShantou University Medical CollegeShantou 515041China

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 *** 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),*** 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 *** then quantified TNF-αandα-SMA mRNA and protein expression levels by qPCR and immunofluorescence(IF)*** interference(RNAi)was used to explore the function of the adipoR *** 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 ***: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)*** detected marked expression of adipoR1,T-cadherin,and calreticulin as well as a moderate presence of adipoR2 in AFs,while no adiponectin was *** adiponectin(gAd)fostered the growth of AFs,and acted in concert with UⅡto induceα-SMA and TNF-αthrough the adipoR1/T-cadherin/calreticulin/AMPK *** AFs,gAd and UⅡsynergistically induced AMPK *** the adventitial inflammation model,APN deficiency up-

关键词： Urotensin II Adiponectin Signal transduction adventitial fibroblast RNA interference(RNAi) Adiponectin-knockout(APN-KO)

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