RutecarpineAttenuatesVascularSmoothMuscleCellCalcificationviaSirt1 / 中山大学学报(医学科学版)

ABSTRACT

@#【Objective】ThisstudyaimstoinvestigatewhetherrutecarpinehasaneffectoncalcificationofVSMCand itsunderlyingmechanism.【Methods】InvitromodelofratVSMCcalcificationwasusedinthisstudy.Rutecarpineat differentconcentrationswasusedtotreatculturedratVSMC.Theexpressionof Runx2，BMP2 and Osterix wasanalyzed byqRT-PCRandmineraldepositionwasdetectedbyalizarinredstaining.Inaddition，weexaminedtheeffectofrutecar⁃ pineonSirtuin-1（Sirt1）expressioninVSMCandtheeffectofSirt1inhibitoronVSMCcalcification.【Results】Alizarinred stainingandcalciumcontentassayshowedthatrutecarpineatdifferentconcentrationssignificantlyreducedcalcificationof ratVSMCinducedbyhighphosphorusandhighcalcium（136±10，75±6，52±6，31±5.29，P<0.05）.Usageofrutecar⁃ pinedecreasedtheactivityofALP，anosteogenicdifferentiationmolecularmarker，anddown-regulatedtheexpressionof Runx2（2.85±0.25，1.75±0.18，1.62±0.13，1.36±0.16，P <0.05），BMP2（3.2±0.32，1.85±0.17，1.65±0.15，1.43± 0.12，P<0.05）andOsterix（2.60±0.27，1.82±0.16，1.55±0.15，1.36±0.17，P<0.05），suggestingthatrutecarpineinhib⁃ ited osteogenic differentiation of VSMC. In addition，high phosphorus and high calcium down-regulated the expres⁃ sionofSirt1inVSMC.qRT-PCRandwesternblotanalysisconfirmedthatrutecarpineup-regulatedtheexpressionof Sirt1atbothmRNA（0.35±0.06，0.75±0.11；0.22±0.08，0.87±0.13，P <0.05）andproteinlevels（0.38±0.09，0.71±0.13，P<0.05）.QuantificationofcalciumcontentanalysisshowedinhibitionofSirt1byEX-527blockedtheinhibitory effectofrutecarpineonVSMCcalcification（138±13，36±7，87±8，P<0.05） .【Conclusion】Wedemonstratethatrutecar⁃ pineattenuatesVSMCcalcificationviaup-regulationofSirt1.