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时间:2018-09-15
《腺相关病毒载体介导的timp1对人退变腰椎间盘髓核细胞蛋白多糖的影响》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、万方数据680中国脊柱脊髓杂志2008年第18卷第9期ChineseJournalofSpineandSpins/Cord.2008,V01.18,No.9腺相关病毒载体介导的TIMPl对人退变腰椎间盘髓核细胞蛋白多糖的影响魏见伟,王德春,胡有谷(青岛大学医学院附属医院脊柱外科266003青岛市)【摘要】目的:探讨腺相关病毒(Adeno—associatedvirus。AAV)载体介导的基质金属蛋白酶组织抑制剂l(tissueinhibitorofmetalloproteinasesl,11MPI)对人退变腰椎问盘髓核细胞
2、的生物学效应。方法:单层培养并鉴定人退变腰椎间盘髓核细胞。采用绿色荧光蛋白标记的腺相关病毒(rAAV2一EGFP)检测其对髓核细胞的转染效率。应用构建的rAAV2一TIMP!转染髓核细胞,通过细胞形态学观察、筠标记氨基酸整合法检测rAAV2-TIMPl对人退变腰椎问盘髓核细胞基质合成的影响。在s5s检测中,以未转染的退变髓核细胞做为正常对照组。结果:光镜下观察所培养的细胞类型以成纤维样细胞为主.Ⅱ型胶原免疫组化和番红O染色鉴定培养细胞为髓核细胞。AAV转染人退变腰椎间盘髓核细胞的转染效率为12%。驾标记整合法检测rAAV2
3、一TIMPl转染组每分钟计数值为341.43±42.85,正常对照组为224.20±29.26.两组间比较差异有统计学意义(P4、iscs:thebio-logicaleffectsoftissueinhibitorofmetalloproteinases1mediatedbyadeno‘‘associatedvirusvec-tor/WElJianwei,WANGDechun,HUYougu//ChineseJournalofSpineandSpinalCord。2008.18(9):680_碣3【Abstract】Objective:Toinvestigatethebiologicaleffectsoftissueinhibitorofmetal5、loproteinase1(TIMPI)mediatedbyadeno—associatedvirus(AAV)onhumandegeneratednucleuspulposusceHs.Method:Humannu-cleuspulposuscellsacquiredfromdegeneratedlumbarintervertebraldiscswereculturedbymonolayercultureandidentified.ThefluorescencelabledAAV(rAAV2一EGFP)wereusedt6、odetectthetransducfionrateofan.cleuspulposuscells.rAAV2-TIMPlwastransducedintohumannucleuspulposuscells,andthenthechangesofextracenularmatrixofnucleuspulposusceHswe陀detectedbycellmorphologyand埯incorporationassay.In35Sincorporationassay。thecontrolgroupWasuntransduc7、eddegeneratednucleuspulposusceUs.Result:ThemajoritytypeofceUswerefibroblast-likecells.Bothimmunohistochemistryofcollagentype1IandSafranineOstainingprovedthatthemonolayerculturedceHswerenucleuspulposusceHs.ThetransductionrateofAAVtothedegeneratednucleuspulposusceUs8、Was12%.In玛incorporationassay.thecountsperminuteofrAAV2-TIMPlandcontrolgroupWas341.43±42.85and224.20±29.26respectively,whichrevealedstatisticaldifference
4、iscs:thebio-logicaleffectsoftissueinhibitorofmetalloproteinases1mediatedbyadeno‘‘associatedvirusvec-tor/WElJianwei,WANGDechun,HUYougu//ChineseJournalofSpineandSpinalCord。2008.18(9):680_碣3【Abstract】Objective:Toinvestigatethebiologicaleffectsoftissueinhibitorofmetal
5、loproteinase1(TIMPI)mediatedbyadeno—associatedvirus(AAV)onhumandegeneratednucleuspulposusceHs.Method:Humannu-cleuspulposuscellsacquiredfromdegeneratedlumbarintervertebraldiscswereculturedbymonolayercultureandidentified.ThefluorescencelabledAAV(rAAV2一EGFP)wereusedt
6、odetectthetransducfionrateofan.cleuspulposuscells.rAAV2-TIMPlwastransducedintohumannucleuspulposuscells,andthenthechangesofextracenularmatrixofnucleuspulposusceHswe陀detectedbycellmorphologyand埯incorporationassay.In35Sincorporationassay。thecontrolgroupWasuntransduc
7、eddegeneratednucleuspulposusceUs.Result:ThemajoritytypeofceUswerefibroblast-likecells.Bothimmunohistochemistryofcollagentype1IandSafranineOstainingprovedthatthemonolayerculturedceHswerenucleuspulposusceHs.ThetransductionrateofAAVtothedegeneratednucleuspulposusceUs
8、Was12%.In玛incorporationassay.thecountsperminuteofrAAV2-TIMPlandcontrolgroupWas341.43±42.85and224.20±29.26respectively,whichrevealedstatisticaldifference
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