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ID:32341562
大小:9.62 MB
页数:36页
时间:2019-02-03
《rock-ⅱ基因表达下调与血管平滑肌细胞迁移的关系》由会员上传分享,免费在线阅读,更多相关内容在学术论文-天天文库。
1、siRNA(so一36433)(P>0.05);(2)themRNAexpressionofROCK-IIinA7r5cellswassignificantlyinhibitedby(CBR0915)(P2、(SC一36433)(P>0。05);(2)theproteinexpressionofROCK-IIinA7r5cellswassignificantlyinhibitedbysiRNA(CBR0915).ThesuppressionofROCK.IImRNAexpressionwas60.I%.Itind沁atedthatsiRNA(CBR09151couldeffectivelyinhibitROCK·IIproteinexpressioninA7r5cells.5。(1)Inthemigrationassay,accordingtothe3、numberofVSMCpassingthroughthemembrane,5ng/mlPDGF-BBwasselectedastheoptimaldosagetoinduceVSMCmigration.(2)ThenumberofVSMCpassingthroughthemembraneintheRNAigroupwas70.2+8.81(n一3)cells,,intheblankcontrolgroupswith59.34-6.22(n=3)cellsandinthelipofectamineTM2000transfectedgroupswi4、th60.14-4.11(n=3)cellsinthe34pmolsiRNAtransfectedgroups.Therewasnostatisticaldifferenc毛amongthethreegroups.Conclusions1.InA7r5cellstransfection,thetransfectionefficiencyoflipofectamine羊鹾2000wasapproximation92%aftera5.hourtransfection.Theoptimaldosageoflipofectamine喇2000was1。75、}llandthefinalconcentrationofsiRNAwasl7nM.2.ItwasindicatedthatsiRNA(CBR0915)couldeffectivelyinhibitROCK-IIinmRNAandproteinexpression.However,siRNA(sc.36433)couldnoteffectivelyinhibtROCK-珏inmRNAandproteinexpression.,3。Down—regulationofROCK-IIgenecouldnotinfluencethenumberofVSM6、Cpassingthroughthemembrane.Therewasnostatisticaldifferenceamongthethreegroups.TheroleofROCK·lIinVSMCmigrationshouldstillbeelucidatedinfuture.Keywords:vascularsmoothmusclecell(VSMC)migrationROCK-IIPlatelet—DerivedGrowthFactor(PDGF)RNAinterference(RNAi)6ROCK—II基因表达下调与血管平滑肌细胞迁移的7、关系硕士研究生:张金超指导教师:高颖教授专业名称:生物化学与分子生物学.^‘·-■一刖菁动脉粥样硬化(Atherosclerosis,AS)及心脑血管疾病是严重威胁人类健康的常见病和多发病,是发达国家人口死亡的主要原因,在我国的死亡率也呈逐年增高的趋势【l】。目前普遍研究认为动脉粥样硬化起始于血管内皮损伤,内皮损伤后血管平滑肌细胞(vascularsmoothmusclecell,VSMC)发生增生及迁移,形成AS斑块。因此,血管平滑肌细胞迁移是AS发生的重要环节。此外,VSMC迁移也是血管成形术后再狭窄发病的一个关键因素。阐明VSMC迁移的发生8、机制对于防止这些增生性血管病具有重要的指导意义。多种细胞外刺激因子对VSMC的迁移有明显作用,如血管活性物质、生长因子和细胞外基质等。血
2、(SC一36433)(P>0。05);(2)theproteinexpressionofROCK-IIinA7r5cellswassignificantlyinhibitedbysiRNA(CBR0915).ThesuppressionofROCK.IImRNAexpressionwas60.I%.Itind沁atedthatsiRNA(CBR09151couldeffectivelyinhibitROCK·IIproteinexpressioninA7r5cells.5。(1)Inthemigrationassay,accordingtothe
3、numberofVSMCpassingthroughthemembrane,5ng/mlPDGF-BBwasselectedastheoptimaldosagetoinduceVSMCmigration.(2)ThenumberofVSMCpassingthroughthemembraneintheRNAigroupwas70.2+8.81(n一3)cells,,intheblankcontrolgroupswith59.34-6.22(n=3)cellsandinthelipofectamineTM2000transfectedgroupswi
4、th60.14-4.11(n=3)cellsinthe34pmolsiRNAtransfectedgroups.Therewasnostatisticaldifferenc毛amongthethreegroups.Conclusions1.InA7r5cellstransfection,thetransfectionefficiencyoflipofectamine羊鹾2000wasapproximation92%aftera5.hourtransfection.Theoptimaldosageoflipofectamine喇2000was1。7
5、}llandthefinalconcentrationofsiRNAwasl7nM.2.ItwasindicatedthatsiRNA(CBR0915)couldeffectivelyinhibitROCK-IIinmRNAandproteinexpression.However,siRNA(sc.36433)couldnoteffectivelyinhibtROCK-珏inmRNAandproteinexpression.,3。Down—regulationofROCK-IIgenecouldnotinfluencethenumberofVSM
6、Cpassingthroughthemembrane.Therewasnostatisticaldifferenceamongthethreegroups.TheroleofROCK·lIinVSMCmigrationshouldstillbeelucidatedinfuture.Keywords:vascularsmoothmusclecell(VSMC)migrationROCK-IIPlatelet—DerivedGrowthFactor(PDGF)RNAinterference(RNAi)6ROCK—II基因表达下调与血管平滑肌细胞迁移的
7、关系硕士研究生:张金超指导教师:高颖教授专业名称:生物化学与分子生物学.^‘·-■一刖菁动脉粥样硬化(Atherosclerosis,AS)及心脑血管疾病是严重威胁人类健康的常见病和多发病,是发达国家人口死亡的主要原因,在我国的死亡率也呈逐年增高的趋势【l】。目前普遍研究认为动脉粥样硬化起始于血管内皮损伤,内皮损伤后血管平滑肌细胞(vascularsmoothmusclecell,VSMC)发生增生及迁移,形成AS斑块。因此,血管平滑肌细胞迁移是AS发生的重要环节。此外,VSMC迁移也是血管成形术后再狭窄发病的一个关键因素。阐明VSMC迁移的发生
8、机制对于防止这些增生性血管病具有重要的指导意义。多种细胞外刺激因子对VSMC的迁移有明显作用,如血管活性物质、生长因子和细胞外基质等。血
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