血管紧张素-1-7对人脐静脉平滑肌细胞钙化影响及其可能机制-effect of angiotensin - 1 - 7 on calcification of human umbilical vein smooth muscle cells and its possible mechanism.docxVIP

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2018-06-03 发布于上海
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血管紧张素-1-7对人脐静脉平滑肌细胞钙化影响及其可能机制-effect of angiotensin - 1 - 7 on calcification of human umbilical vein smooth muscle cells and its possible mechanism.docx

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血管紧张素-1-7对人脐静脉平滑肌细胞钙化影响及其可能机制-effect of angiotensin - 1 - 7 on calcification of human umbilical vein smooth muscle cells and its possible mechanism

randomlydividedinto6groups(n=6):thecontrolgroup(CON),calcificationgroup(CAL),pureangiotensin-(1-7)group[Ang-(1-7)group],angiotensin-(1-7)withhigh-doseincalcifiednutrientsolution[Ang-(1-7)Hgroup],angiotensin-(1-7)withmoderatedoseinthecalcifiednutrientsolution[Ang-(1-7)MGroup],angiotensin-(1-7)withlow-doseincalcifiednutrientsolutiongroup[Ang-(1-7)LGroup].2.2.AnaLysis:①CalciumdepositioninHUSMCsweredetectedbyVonKossastaining.②VonKossastainingcalculatedbymicro-imageanalysissystem.③Theactivitiesofalkalinephosphatase(ALP)inHUSMCswereverifiedbypheny1diphosphate-2-sodium;thetotalcalciumcontentsinHUSMCswerecalculatedbyultravioletspectrophotometry.④ActivityofTGFβ1inthenutrientsolutionwereanalysisedbyenzyme-linkedimmunosorbentassay,determineingthechangeofthelevelofTGFβ1inHUSMCs.⑤Theexpressionofcorebindingfactorα1(cbfα1)inHUSMCswereanalyzedbyWestern-blot.⑥Osteocalcin(OC)contentsinthenutrientsolutionweredeterminedbyradioimmunoassay.Results:HUSMCscalcificationwerereproducedsuccessfully:usingβ-glycerolphosphate-inducedcalcificationinvitroHUSMCs.AfterVonKossastaining,alargenumberofdaisy-shapedHUSMCsweresignificantlydetectedingroup12mmol/L.Therewerebrownnodulesamongsmoothmusclecell.ComparedwithCONgroup,Micro-imageanalysisofquantitativecalculationsshowthatthepercentageofcalcifiedcellsin12mmol/Lgroupweresignificantlyincreased(P0.05),10mmol/Lgroup,14mmol/Lgroup,16mmol/Lgroupwerealsoincreased(respectivelyP0.01);andcellcalciumcontentandALPlevelswereincreasedrespectively(P0.001).But12mmol/Lgroupwassignificantlyincreased.Angiotensin-(1-7)effectsonHUSMCscalcification①Ang-(1-7)cansignificantlyreducethecalcificationoftheHUSMCs.ComparedwithCALgroup,VonKossastainingofAng-(1-7)Hgroup,Ang-(1-7)Mgroup,Ang-(1-7)Lgroupshowedbrownnodulesweresignificantlyreduced(P0.001);ComparedwithCALgroup,thepercentageofpositivecellsincalcification,thecalciumcontentandALPlevelsofAng-(1-7)Hgroup,Ang-(1-7)Mgroup,Ang-(1-7)Lgroupwassignificantlyreducedrespectively(P0.001);②ComparedwiththeCALgroup,TGFβ1levelsofAng-(1-7)Hg