靜脈移植骨髓間充質(zhì)干細(xì)胞促進(jìn)脊髓損傷后GDNF基因的表達(dá).doc

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1、靜脈移植骨髓間充質(zhì)干細(xì)胞促進(jìn)脊髓損傷后GDNF基因的表達(dá)【摘要】　?。勰康模萏接懡?jīng)靜脈移植骨髓間充質(zhì)干細(xì)胞(MSCs)對(duì)大鼠脊髓損傷(SCI)后膠質(zhì)細(xì)胞源性神經(jīng)營(yíng)養(yǎng)因子(GDNF)表達(dá)的影響。［方法］MSCs提取自成年Wistar大鼠的股骨干骨髓，經(jīng)原代培養(yǎng)、鑒定及5-溴脫氧尿嘧啶核苷(Brdu)核標(biāo)記。以改良Allen′s打擊裝置制作大鼠T10節(jié)段脊髓損傷(SCl)模型，于傷后立即縫合切口并經(jīng)尾靜脈注射移植MSCs。實(shí)驗(yàn)共分為3組：MSCs尾靜脈移植組(A組)、生理鹽水注射組(B組)、正常對(duì)照組(C組)。術(shù)后不

2、同時(shí)間點(diǎn)應(yīng)用免疫組化法和逆轉(zhuǎn)錄聚合酶鏈反應(yīng)法(RTPCR)觀察MSCs移植后的存活狀態(tài)以及不同時(shí)間點(diǎn)GDNF基因的表達(dá)變化。［結(jié)果］免疫組化結(jié)果：MSCs經(jīng)尾靜脈移植后在損傷脊髓平面可以檢測(cè)到遷移及存活，標(biāo)記細(xì)胞呈棕黃色的核標(biāo)記，以損傷區(qū)域?yàn)槎嗖⑾蛑車w移，移植術(shù)后第14d，A組Brdu陽(yáng)性細(xì)胞較多，最遠(yuǎn)于距離損傷區(qū)2.5cm處可檢測(cè)到。B及C組則均未檢測(cè)到陽(yáng)性細(xì)胞。RTPCR結(jié)果：移植術(shù)后第1、3、5d，A組表達(dá)量呈逐漸升高趨勢(shì)，B組呈一過(guò)性表達(dá)升高，C組無(wú)變化。各時(shí)間點(diǎn)A組GDNFmRNA的表達(dá)量明顯高于

3、B組，P<0.05。免疫組化結(jié)果：移植術(shù)后第7、14、28dGDNF的表達(dá)量明顯高于B組，P<0.05。［結(jié)論］骨髓間充質(zhì)干細(xì)胞經(jīng)尾靜脈移植后可遷移至脊髓損傷區(qū)域，并上調(diào)膠質(zhì)細(xì)胞源性神經(jīng)營(yíng)養(yǎng)因子基因的表達(dá)，是該移植方式修復(fù)大鼠脊髓損傷的機(jī)制之一?！娟P(guān)鍵詞】脊髓損傷骨髓間充質(zhì)干細(xì)胞膠質(zhì)細(xì)胞源性神經(jīng)營(yíng)養(yǎng)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