Uncontrolled proliferation,migration and phenotypic switching of vascular smooth muscle cells(VSMCs)are im-portant steps in the development and progression of aortic dissection(AD).The function and potential mechanism of miR-335-5p in the pathogenesis of AD are explored in this study.Specifically,the biological function of miR-335-5p is explored in vitro through CCK-8,Transwell,immunofluorescence,EdU,wound-healing,RT-qPCR and western blotting assays.In addition,an AD model induced by angiotensin Ⅱ is used to investigate the function of miR-335-5p in vivo.A dual-luciferase assay is performed to verify the targeting relationship between miR-335-5p and specificity protein 1(SP1).Experiments involving the loss of SP1 function are performed to demonstrate the function of SP1 in the miR-335-5p-mediated regulation of human aortic-VSMCs(HA-VSMCs).AD tissues and platelet-derived growth factor BB(PDGF-BB)-stimulated HA-VSMCs show significant downregulation of miR-335-5p expression and upre-gulated SP1 expression.Overexpression of miR-335-5p effectively suppresses cell proliferation,migration and synthetic phenotype markers and enhances contractile phenotype markers induced by PDGF-BB treatment.Ad-ditionally,SP1 is identified as a target gene downstream of miR-335-5p,and its expression is negatively correlated with miR-335-5p in AD.Upregulation of SP1 partially reverses the inhibitory effect of miR-335-5p on HA-VSMCs,whereas the downregulation of SP1 has the opposite effect.Furthermore,Ad-miR-335-5p clearly suppresses aorta dilatation and vascular media degeneration in the AD model.Our results suggest that miR-335-5p inhibits HA-VSMC proliferation,migration and phenotypic switching by negatively regulating SP1,and indicate that miR-335-5p may be a potential therapeutic target in AD.

Runwei Ma;Dayong Zhang;Yi Song;Jichang Kong;Chunjie Mu;Pin Shen;Wenting Gui

Department of Cardiovascular Surgery,Fuwai Yunnan Cardiovascular Hospital,Kunming 650102,China;Department of Cardiovascular Surgery,Guangyuan Central Hospital,Guangyuan 628099,China;Department of Extracorporeal Circulation,Fuwai Yunnan Cardiovascular Hospital,Kunming 650102,China

生物化学与生物物理学报（英文版）

[1]Runwei Ma;Dayong Zhang;Yi Song;Jichang Kong;Chunjie Mu;Pin Shen;Wenting Gui-.miR-335-5p regulates the proliferation,migration and phenotypic switching of vascular smooth muscle cells in aortic dissection by directly regulating SP1)[J].生物化学与生物物理学报（英文版）,2022(07):961-973

Uncontrolled,upre,gulated

miR,5p,regulates,proliferation,migration,phenotypic,switching,vascular,smooth,muscle,cells,aortic,dissection,by,directly,regulating,SP1,VSMCs,are,portant,steps,development,progression,AD,function,potential,mechanism,pathogenesis,explored,this,study,Specifically,biological,vitro,through,CCK,Transwell,immunofluorescence,EdU,wound,healing,qPCR,western,blotting,assays,In,addition,model,induced,angiotensin,used,investigate,vivo,dual,luciferase,performed,verify,targeting,relationship,between,specificity,protein,Experiments,involving,loss,demonstrate,mediated,human,HA,tissues,platelet,derived,growth,BB,PDGF,stimulated,show,significant,downregulation,Overexpression,effectively,suppresses,synthetic,phenotype,markers,enhances,contractile,treatment,Ad,ditionally,identified,downstream,negatively,correlated,Upregulation,partially,reverses,inhibitory,whereas,has,opposite,Furthermore,clearly,aorta,dilatation,degeneration,Our,results,suggest,that,inhibits,indicate,may,therapeutic

0.461842