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时间:2020-05-04
《OPN影响小鼠MSCs迁移及其相关分子机制研究-论文.pdf》由会员上传分享,免费在线阅读,更多相关内容在应用文档-天天文库。
1、重庆医学2014年2月第43卷第4期391·论著·OPN影响小鼠MSCs迁移及其相关分子机制研究李薇,王文平,陈亮,王珍祥△,李世荣(第三军医大学附属西南医院整形外科,重庆400038)摘要:目的探讨外源骨桥蛋白(OPN)对小鼠骨髓间充质干细胞(MSCs)迁移能力的影响。方法采用OPN一/一小鼠和野生型C57小鼠,进行MSCs的原代分离培养,流式细胞术鉴定分拣MSCs细胞传代培养;Transwell迁移实验检测0PN是否能够诱导MSCs定向迁移;蛋白免疫印迹法(westernblot)检测OPN、CD44
2、和Integrin131相关蛋白的表达变化。结果传代后的细胞形态符合MSCs特征,细胞表达cD44和CD105,但不表达CD34,符合MSCs表面标记抗原的一般规律。O.5/~g/mL的OPN能增加体外MSCs的细胞迁移,而野生型C57小鼠MSCs细胞迁移为最多。同时,这一趋势与OPN作用时间正相关,与OPN一/一小鼠相比,差异均有统计学意义(JP<0.05)。0.5g/mL重组0PN蛋白量明显增加,但仍低于野生型C57小鼠,差异有统计学意义(P3、/mL重组OPN组CD44、Integrin81蛋白表达增多,差异有统计学意义(P<0.01);而野生型C57小鼠细胞CD44、IntegrinB1蛋白表达最多,差异有统计学意义(P<0.01)。结论OPN可以通过上调CD44、Integrin61的表达,促进MSCs定向迁移。关键词:骨桥蛋白质;骨髓间充质干细胞;细胞运动;分子机制doi:10.3969/j.issn.1671-8348.2014.04.003文献标识码:A文章编号:167I-8348(2O14)O4一O391一O3Studyonthee4、ffectsofOPNonmigrationofMSCsanditsmolecularmechanisminmiceL,WangWenping,ChenLiang,WangZhenxiang,LiShirong(DepartmentofPlasticSurgery,SouthwestHospital,ThirdMilitaryMedicalUniversity,Chongqing400038,China)Abstract:0bjectiveToinvestigatetheeffectsofexogenou5、sosteopontin(OPN)themigrationofbonemarrowmesenchymalstemcells(MSCs)inmice.MethodsUsingtheOPN一/一andwild—typeC57mice,theMSCswereisolatedandcultured.Thesecellswereanalysedandsortedbyflowcytometry.Then,TranswellmigrationassaydetectedwhetherCIPNcaninducethemig6、rationoftheseMSCs.Finally,themethodofWesternblotwasusedtodetectthechangesofexpressionofOPN,CD44andIntegrin131pro—teins.ResultsThemorphologyandfeaturesofMSCswereprovedcorrectlyon3passages,whichthesecellsexpressedtheproteinsofCD44andCD105,butnotCD34.Thesurf7、acemarkerantigenswereaccordedwiththegenera1featuresofMSCs.Comparedwith0PN一/一mice.theMSCssignificantlyincreasedcellmigrationingroupsof0.5~g/mLOPNandwildtypeC57mice,whichwaspositivelyrelatedtothetimesusingOPN.UsingrecombinantOPNprotein(0.5/,g/mL),expression8、ofrelatedproteinswassignifi—cantlyincreased.butstilllowerthanthegroupofwildtypemice(P<0.01).ComparedwithOPN一/一mice,expressionofOPN,CD44.andIntegrint31proteinincreasedsignificantlydifferent(P<0.01)ingroupsof0.5/,g/mL
3、/mL重组OPN组CD44、Integrin81蛋白表达增多,差异有统计学意义(P<0.01);而野生型C57小鼠细胞CD44、IntegrinB1蛋白表达最多,差异有统计学意义(P<0.01)。结论OPN可以通过上调CD44、Integrin61的表达,促进MSCs定向迁移。关键词:骨桥蛋白质;骨髓间充质干细胞;细胞运动;分子机制doi:10.3969/j.issn.1671-8348.2014.04.003文献标识码:A文章编号:167I-8348(2O14)O4一O391一O3Studyonthee
4、ffectsofOPNonmigrationofMSCsanditsmolecularmechanisminmiceL,WangWenping,ChenLiang,WangZhenxiang,LiShirong(DepartmentofPlasticSurgery,SouthwestHospital,ThirdMilitaryMedicalUniversity,Chongqing400038,China)Abstract:0bjectiveToinvestigatetheeffectsofexogenou
5、sosteopontin(OPN)themigrationofbonemarrowmesenchymalstemcells(MSCs)inmice.MethodsUsingtheOPN一/一andwild—typeC57mice,theMSCswereisolatedandcultured.Thesecellswereanalysedandsortedbyflowcytometry.Then,TranswellmigrationassaydetectedwhetherCIPNcaninducethemig
6、rationoftheseMSCs.Finally,themethodofWesternblotwasusedtodetectthechangesofexpressionofOPN,CD44andIntegrin131pro—teins.ResultsThemorphologyandfeaturesofMSCswereprovedcorrectlyon3passages,whichthesecellsexpressedtheproteinsofCD44andCD105,butnotCD34.Thesurf
7、acemarkerantigenswereaccordedwiththegenera1featuresofMSCs.Comparedwith0PN一/一mice.theMSCssignificantlyincreasedcellmigrationingroupsof0.5~g/mLOPNandwildtypeC57mice,whichwaspositivelyrelatedtothetimesusingOPN.UsingrecombinantOPNprotein(0.5/,g/mL),expression
8、ofrelatedproteinswassignifi—cantlyincreased.butstilllowerthanthegroupofwildtypemice(P<0.01).ComparedwithOPN一/一mice,expressionofOPN,CD44.andIntegrint31proteinincreasedsignificantlydifferent(P<0.01)ingroupsof0.5/,g/mL
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