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1、山東醫(yī)藥2014年第54卷第29期外源性神經(jīng)生長因子對雪旺細胞中NF—KBmRNA表達的影響辛林偉。王梨明，唐際存，李朝旭，李強(桂林醫(yī)學院附屬醫(yī)院，廣西桂林540001)摘要：目的觀察外源性神經(jīng)生長因子(NGF)對雪旺細胞(SCs)中核轉(zhuǎn)錄因子t(B(NF—KB)mRNA表達的影響。方法體外培養(yǎng)SCs，用S-100免疫組化染色鑒定細胞純度。取第4代優(yōu)質(zhì)細胞，隨機分為實驗組(NGF)、阻斷組(NGF+anti．NGFR)、對照組(PBS)，提取各組總mRNA，分別用紫外分光光度計、1．5％瓊脂糖凝膠電泳觀察結(jié)果，通過實時熒光定量PCR檢測NF—KBmRNA的表達。結(jié)果體外
2、培養(yǎng)的SCs與倒置顯微鏡下觀察到的活體細胞形態(tài)一致，S-100免疫組化染色顯示，SCs純度達到91％。紫外分光光度計觀察A／A為1．8～2．2，1．5％瓊脂糖凝膠電泳可見28、18S兩條清晰條帶，證實提取的mRNA質(zhì)量良好。實時熒光定量PCR檢測發(fā)現(xiàn)，與對照組、阻斷組相比，實驗組NF．KBmRNA的表達顯著增高(P均<0．01)。結(jié)論外源性NGF可以上調(diào)SCs中NF—KBmRNA的表達。關(guān)鍵詞：神經(jīng)生長因子；雪旺細胞；核轉(zhuǎn)錄因子KBdoi：10．3969／j．issn．1002—266X．2014．29．009中圖分類號：R741文獻標志碼：A文章編號：1002-266X
3、(2014)29-0024-03Influenceofexogenousnervegrowthfactoronexpression0fNF．KBmRNAofSchwanncellsXINLin—wei，WANGLi—ming，TANGji—curt，LIZhao—“，LIQiang(AfiliatedHospitalofGuilinMedicalCollege，Guilin540001，China)Abstract：ObjectiveToobservetheinfluenceofexogenousnervegrowthfactor(NGF)ontheexpressiono
4、fnucleartranscriptionfactor—KB(NF—KB)mRNAofSchwanncells(SCs)．MethodsSchwanncellswereisolatedandculturedinvitro．ThepositivecellswerecountedbyusingS-100immunohistochemicalstaining．Thefourthgenerationofhigh—qualitycellswererandomlydividedintotheexperimentalgroup(NGF)，blockinggroup(NGF+anti-N
5、GFR)andcontrolgroup(PBS)．ThetotalmRNAineachgroupwereextractedandobservedbyultravioletspectrophotometerand1．5％agarosegelelectrophoresis，respectively，andtheexpressionofNF—KBmRNAweredetectedbyreal—timefluorescencequantitativePCR．ResultsThelivingcellmorphologiesofSCsculturedinvitrowereconsist
6、entwiththoseobservedunderinvertedmi-croscope．S-100immunohistochemicalstainingshowedthatthepurityofSCswas91％．Ultravioletspectrophotometerob-servedthatA260／A280was1．8-2．2，1．5％agarosegelelectrophoresisshowedthattherewere28and18Sclearbands，whichconfirmedthequalityofextractedmRNAwasgood．Realti
7、mefluorescencequantitativePCRindetectionofexpres—sionofNF—KBmRNAshowedthatwhencomparedwiththecontrolgroupandblockinggroup，theexpressionofNF—KBmR—NAintheexperimentalgroupWassignificantlyincreased(allP<0．01)．ConclusionsExogenousNGFcanup—regulatetheexpressionofNF-KBmRN
