静脉移植骨髓间充质干细胞促进脊髓损伤后GDNF基因的表达.doc

《静脉移植骨髓间充质干细胞促进脊髓损伤后GDNF基因的表达.doc》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、静脉移植骨髓间充质干细胞促进脊髓损伤后GDNF基因的表达【摘要】　　［目的］探讨经静脉移植骨髓间充质干细胞(MSCs)对大鼠脊髓损伤(SCI)后胶质细胞源性神经营养因子(GDNF)表达的影响。［方法］MSCs提取自成年Wistar大鼠的股骨干骨髓，经原代培养、鉴定及5-溴脱氧尿嘧啶核苷(Brdu)核标记。以改良Allen′s打击装置制作大鼠T10节段脊髓损伤(SCl)模型，于伤后立即缝合切口并经尾静脉注射移植MSCs。实验共分为3组：MSCs尾静脉移植组(A组)、生理盐水注射组(B组)、正常对照组(C组)。术后不

2、同时间点应用免疫组化法和逆转录聚合酶链反应法(RTPCR)观察MSCs移植后的存活状态以及不同时间点GDNF基因的表达变化。［结果］免疫组化结果：MSCs经尾静脉移植后在损伤脊髓平面可以检测到迁移及存活，标记细胞呈棕黄色的核标记，以损伤区域为多并向周围迁移，移植术后第14d，A组Brdu阳性细胞较多，最远于距离损伤区2.5cm处可检测到。B及C组则均未检测到阳性细胞。RTPCR结果：移植术后第1、3、5d，A组表达量呈逐渐升高趋势，B组呈一过性表达升高，C组无变化。各时间点A组GDNFmRNA的表达量明显高于

3、B组，P<0.05。免疫组化结果：移植术后第7、14、28dGDNF的表达量明显高于B组，P<0.05。［结论］骨髓间充质干细胞经尾静脉移植后可迁移至脊髓损伤区域，并上调胶质细胞源性神经营养因子基因的表达，是该移植方式修复大鼠脊髓损伤的机制之一。【关键词】脊髓损伤骨髓间充质干细胞胶质细胞源性神经营养因子　　Effectsofmesenchymalstemcellsintravenoustransplantationontheglialcelllinederivedneurotrophicfactora

4、fterthespinalcordinjuryofrats∥　　Abstract：［Objective］Toobservetheeffectsofmesenchymalstemcells(MSCs)injectionfromtailveinontheexpressionofglialcelllinederivedneurotrophicfactor(GDNF)afterthespinalcordinjury(SCI)ofrats.［Method］MSCswereculturedfromthethighboneo

5、fadultwistarrats.TheT10levelspinalcordinjurywasexecutebythemodifyAllen'sdevice.TheMSCslabeledbybromodeoxyuridineweretransplantedintotheveinoftailbyinjectionimmediatelyafterspinalcordinjury.AdultWistarratswererandomlydividedintothreegroups:SCIcuredwithMSCstra

6、nsplantation(groupA),SCIreceivednormalsaline(groupB),andcontrolgroup(groupC).ThenMSCsweredetectedandtheexpressionsofGDNFofthelesionandneighborareaswereexaminedbyReversetranscriptionpolymerasechainreaction(RTPCR)andimmunohistochemistry.［Result］Immunohistochem

7、istry:MSCslabeledbybromodeoxyuridinecouldbedetectedintheinjuriedspinalcord　aftertransplantation.ItwaslocatedatthenucleolusofMSCs.Thecellssurvivedandmigratedrostrallyandcaudallyfromtheinjectionsites.Withthetimepassed,thenumberofMSCslabeled5with　bromodeoxyuridi

8、newasdecreased.RT-PCR:TheexpressionofGroupAwasincreased　onthefollowed　day,andtheexpressionofGroupBwasincreasedonthe1stdaybutdecreasefromthe5thdaythenon.comparedwithgroupB,transplantationo