外源性神经生长因子对雪旺细胞中NF-κB mRNA 表达的影响.pdf

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1、山东医药2014年第54卷第29期外源性神经生长因子对雪旺细胞中NF—KBmRNA表达的影响辛林伟。王梨明，唐际存，李朝旭，李强(桂林医学院附属医院，广西桂林540001)摘要：目的观察外源性神经生长因子(NGF)对雪旺细胞(SCs)中核转录因子t(B(NF—KB)mRNA表达的影响。方法体外培养SCs，用S-100免疫组化染色鉴定细胞纯度。取第4代优质细胞，随机分为实验组(NGF)、阻断组(NGF+anti．NGFR)、对照组(PBS)，提取各组总mRNA，分别用紫外分光光度计、1．5％琼脂糖凝胶电泳观察结果，通过实时荧光定量PCR检测NF—KBmRNA的表达。结果体外

2、培养的SCs与倒置显微镜下观察到的活体细胞形态一致，S-100免疫组化染色显示，SCs纯度达到91％。紫外分光光度计观察A／A为1．8～2．2，1．5％琼脂糖凝胶电泳可见28、18S两条清晰条带，证实提取的mRNA质量良好。实时荧光定量PCR检测发现，与对照组、阻断组相比，实验组NF．KBmRNA的表达显著增高(P均<0．01)。结论外源性NGF可以上调SCs中NF—KBmRNA的表达。关键词：神经生长因子；雪旺细胞；核转录因子KBdoi：10．3969／j．issn．1002—266X．2014．29．009中图分类号：R741文献标志码：A文章编号：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