腺病毒载体搭载的timp

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1、腺病毒载体搭载的TIMP.L.编辑。作者：XIONGLi-ming熊蠡茗,GUOBing郭兵,SHAOZeng-lRAd66(第2组)以及1.0×1010OPU/mlRAdTIMP-3(第3、4、5组)各25μl。各组分别于注射后2、2、1、2、4周收集标本，X-gal染色检测转染效率，RT-PCR检测TIMP-3及聚集蛋白聚糖核心蛋白的表达，TIMP-3及H型胶原免疫组化染色、藏红O-快绿染色及Tunel染色考察转染对椎间盘基质的调控作用。[结果](1)RAdTIMP-3经扩增和纯化后浓度可达1.9×10TMOPU/ml。(2)RT-PCR显示，3、4、5组TIMP-3基因表达均较1

2、、2组明显增加，3、4、5组核心蛋白基因表达较1、2组略有增强。(3)Tunel染色显示各组凋亡细胞比例无明显差异。(4)第5组藏红O-快绿染色及II型胶原免疫组化染色染色强度均好于1、2组。[结论]RAdTIMP-3能够广泛而安全地在兔椎间盘内表达，并改善椎间盘基质成分的质和量，具备用于椎间盘退变治疗的潜力。【关键词】组织金属蛋白酶抑制因子-3;椎间盘;聚集蛋白聚糖;Ⅱ型胶原　　Abstract：[Objective]ToinvestigatetheinfluenceofrebinantadenoviruscarryingtissueinhibitorofmetalloprotEin

3、ase-3(RAdTIMP-3)onthemainpositionsofrabbitsintervertebraldiscsandtoassessitspotentialintreatmentforintervertebraldiscdegeneration.[Method]RadTIMP-3andemptyadenovimsvectorlydividedinto5groups.And25μlofvariousreagentsalsalineingroup1,1.0×1010OPU/mlofRAd66inGroup2,and1.0×1010OPU/mlofRAdTIMP-3ingrou

4、p3,4and5.TheintervertebraldiscsofeachgroupmunohistochemicalstainingforTIMP-3andtypeI!collagenandSafraninO-Fastgreenstaininglafterpropagationandpurification.(2)RT-PCRshomunohistochemicalstainingfortypeIIcollagenofgroup5provethequantityandqualityofmatrix.Ithasthepotentialtobeusedintreatmentforinte

5、rvertabraldiscdegeneration.　　Keyetalloproteinases-3;intervertebraldisc;aggrecan;typeⅡcollagenDegenerationofintervertebraldisc(IVD)isthemaincauseofthemonspiuaediseasessuchasprolapseofintervertebraldiscandspinalstenosisetc.Itedicalresources.IVDisamatrix-basedtissue,thedegenerationofthemainmatrixpo

6、sition:aggrecanisthekeypointandthefirstimportantperformanceofoeegeneratinofIVD.Sotocontrolandprotectthematrixisasignificantdirectioninthedegeueratiostimportantproteaseinthedegradationprocessofaggrecan.Thetissueinhibitorofmetalloproteinase(TIMPs)familykeepingtheimportantbalanceofmatrixmetabolism,

7、arethenaturalinhibitorsofaggrecanaseandMMPs.TIMP-3ofthefamilyshoesmorethanTIMP-1[1]usedintherecentresearchofgenetherapyofdegeuorativeIVD.Theoretically,itcanplayatherapyroleinraisingthequalityandamountoftheintervertebralmatri