shrna慢病毒转染串珠素对大鼠硬膜外瘢痕中成纤维细胞增殖能力影响论文

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1、Abs仃act0bjectiveAfterlaminectomyepiduratscarformationisanormalphysiologicalprocessoftissuerepair,andisalsooneofthelumbarspinalpostoperativecomplications，scartissueandthenucleusotepiduraladhesionscallleadtoclinicalpathologicalsymptomsoIrecurrence，andinhibi

2、ttheexcessiveproliferationofscartissueisoneottheimportantmethodstopreventlumbarsurgeryfailure．Thisstudyistoresearchthefunctionofexpressi_onofperlecanshRNAtransfectiononepiduralcica廿ixfibroblast．MethodsOperatingon30Wstarmoussetoestablishtheformationofconep

3、lateresectionandtraining4weeksaftersurgery．Extractionorganizationofepiduralscarinthefourthweek．Culturefibroblastsofepiduralscartissuebytissueadherentanddividedintothreegroups：shRNAgroup、GFPgroupandcontrolgroup．DetecttheexpressionsofperlecanmRNAandproteina

4、ndcellproliferationbyWestern—blot、realtimequantitativePCRandMTTafterfibroblasttransfected．．ResultPerlecanmRNAandproteinshowedhighexpressioninthecontotandGFPgroupsbutlowexpressionintheshRNAgroup，withsignificantdifferencerespectivelybetweentheshRNAgroupando

5、ther2groups．Therewasnosignificantdifferencebetweenthe3groupsintheopticaldensityattheday(P>O．05)．On24to96hourthecellsinthecontrolandGFPgroupgrewnormallywhilethecellsintheshRNAgroupproliferatedinaweakmanner．ThetransfectedcellsintheshRNAgroupshowedasignifica

6、ntlyreducedproliferationratecomparewiththeother2groups(P

7、words]RNA；Transfection；fibroblast；epiduralscar；lentivirus引言第1章材料与方法目录⋯⋯⋯⋯⋯⋯⋯⋯．31．1仪器、材料与实验试剂⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯．31．1．1实验动物⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯．31．1．2细胞培养主要试剂⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯．31．1．3细胞培养主要溶液配制⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯．31．1．4目的细胞慢病毒转染主要仪

8、器⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯41．1．5目的细胞慢病毒转染主要试剂⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯41．1．6目的细胞慢病毒转染主要溶液配制⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯41．1．7RT—PCR主要仪器⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯41．1．8RT—PCR主要试剂⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯51．1．9RT—PCR主要溶液配制⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯