血管紧张素 1 7对人脐静脉平滑肌细胞钙化的影响及其可能机制-effect of angiotensin 17 on calcification of human umbilical vein smooth muscle cells and its possible mechanism.docxVIP

蛋白表达。④放射免疫技术检测培养液中骨钙素（OC）含量，观察HUSMCs骨钙素变化。结果：1.HUSMCs钙化模型复制不同浓度β-GP诱导体外HUSMCs钙化，VonKossa染色结果显示12mmol/L组大量HUSMCs呈菊花状聚集生长，细胞聚集处可见棕褐色钙结节，显微图像分析系统定量计算结果发现12mmol/L组与CON组、10mmol/L组、14mmol/L组、16mmol/L组相比，钙化细胞百分比明显增加，细胞钙含量和ALP活性均显著升高，有显著性差异。2.血管紧张素-（1-7）对HUSMCs钙化影响①Ang-(1-7)可明显减少HUSMCs聚集生长，VonKossa染色显示细胞聚集处棕褐色钙结节较CAL组明显减轻（P0.01)，Ang-(1-7)高、中、低剂量组钙化细胞百分比、细胞钙含量和ALP活性较CAL组明显降低（均为P0.01)。②Ang-(1-7)高、中、低剂量组与CAL组相比细胞TGFβ1水平、cbfα1表达、骨钙素含量均降低（分别P0.01)。结论：1．12mmol/Lβ-GP刺激HUSMCs10天，可成功复制体外HUSMCs钙沉积模型。2．采用显微图像分析系统可定量计算出VonKossa染色钙沉积情况，可作为鉴定VonKossa染色钙化的辅助方法。。3、Ang-(1-7)可以减轻HUSMCs钙化程度。4、Ang-(1-7)通过影响TGFβ1－Smads－Cbfα1信号通路级联效应，减弱信号通路中关键信号分子TGFβ1、Cbfα1表达，发挥延缓HUSMCs钙化的作用。关键词血管紧张素-（1-7），血管钙化，转化生长因子β1，核心结合因子1，骨钙素TheEffectsofAngiotensin-(1-7)onVascularCalcificationinHumanUmbilicalVeinSmoothMuscleCellandtheUnderlyingMechanismABSTRACTObjectives:Inthepresentstudy,weestablishthehumanumbilicalveinsmoothmusclecell(HU--SMCs)calcificationmodelwhichinducedbyβ-Glycerophosphate,andobservedtheeffectofAngiotensin-(1-7)[Ang-(1-7)]invascularcalcification,investigatingthepossiblemech--anismswhichwasAng-(1-7)delayorinhibittheHUSMCscalciumdeposition.Method:ReproductionofvascularcalcificmodelUsingβ-glycerolphosphate(β-GP)stimulatethehumanumbilicalveincellswhichculturedinvitro.Thecellswererandomlydividedinto5groups(n=6):thecontrolgroup(CON),10mmol/L,12mmol/L,14mmol/L,16mmol/Lβ-GP(I.e:10mmol/Lgroup,12mmol/Lgroup,14mmol/Lgroup,16mmol/Lgroup).CalciumdepositioninHUSMCsweredetectedbyVonKossastaining;andcalculatedVonKossastainingbymicro-imageanalysissystem;theactivitiesofalkalinephosphatase(ALP)inHUSMCswereverifiedbypheny1diphosphate-2-sodium;thetotalcalciumcontentswerecalculatedbyultravioletspectrophotometry.Angiotensin-(1-7)effectsonHUSMCscalcificationInterventedangiotensin-(1-7)incalcifiednutrientsolution.,andexperimentalgroupwererandomlydividedinto6groups(n=6):thecontrolgroup(CON),calcificationgroup(CAL),pureangiotensin-(1-7)group[Ang-(1-7)group],angiotensin-(1-7)withhigh-doseincalcifiednutrientsolution[Ang-(1-7)Hg