甲氨蝶呤对早期神经胚基因表达的影响-论文

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1、·5058·现代生物医学进展www．shengwuyixue．eomProgressinModemBiomedicineVoL14NO．26SEP．2014doi：10．13241／j．cnki．pmb．2014．26．015甲氨蝶呤对早期神经胚基因表达的影响术季成王秀伟王建华官臻朱智强谢秋张霆牛勃1,2A．(1山西医科大学基础医学院山西太原030001；2首都儿科研究所儿童发育营养组学北京市重点实验室北京北京100020)摘要目的：利用甲氨蝶呤(methotrexate，MTX)干预孕鼠，探讨MTX对早期神经胚基因表达的影响。方法：用MTX(4．

2、5mg／kg体重)干预孕鼠，通过NimbleGene表达谱芯片、Realtime—PCR及免疫组化等方法进行差异表达基因的筛选和验证。结果：MTX处理后神经管畸形(NTDs)发生率为32．1％。表达谱芯片筛选出166个差异表达基因，其中4个凋亡相关基因(Endog，Trp53，Casp3，Bax)均表现为上调(foldchange>1．5，P<0．05)，3个增殖相关基因(Ptchl，Pla2g4a，Foxg1)均表现为下调(foldchange<0．67，P<0．05)；NTDs胚胎神经上皮Caspase一3表达显著升高(P<0．05)，phos

3、pho-histoneH3(pH3)表达显著降低(P<0．05o结论：MTX影响了早期神经胚的基因表达，尤其是引起了凋亡、增殖相关基因表达的异常，这可能在叶酸缺乏引起NTDs发生的相关机制之一。关键词：神经管畸形；甲氨蝶呤；凋亡；增殖中图分类号：Q95—3，R722文献标识码：A文章编号：1673—6273(2014)26—5058·05AlterationofGeneExpressionbyMethotrexateExposureatEarlyNeurulation*JICheng~,WANGXiu—wef,WANGJian-hu．GUANZhe

4、n2,ZHUZhi-qiange,XIEQiue,ZHANGTinge,NlUbd世(1SchoolofBasicMedicalSciences,ShanxiMedicalUniversi~Taiyuan,Shanxi,030001，China；2CapitalInstituteofPedia~'cs,Beijing,100020,China)ABSTRACTObjective：Weusedmethotrexate(MTX)interventioninpregnantC57BL／6Jmicetoinvestigatethealterationofg

5、eneexpressionatearlyneurulation．Methods：ThediferentialgeneexpressionswerestudiedbyMicroarray，RT—PCRandInlnlunohistochemicalassaysinNTDembryosinducedbyMTX(4．5mg／kgbodyweight．Results：Resultsshowedthat32．1％ofNTDswasdevelopedbythetreatmentofMTX．Microarrayindicatedthat166genesweres

6、ignificantlydiferentbetweencontrolandNTDmice，including4apoptosis—relatedgenes(Endog，Trp53，Casp3，Bax)and3proliferation-relatedgenes(Ptchl，Pla2g4a，Foxg1)．LevelsofEndog，Trp53，Casp3，Bax(foldchange>1．5)wereup·regulatedbutPtchl，Pla2g4a，Foxgl(foldchange<0．67)weredown—regulated(P<0．05

7、)．Expressionofcaspase一3wassignificantlyenhanced(P<0．05)whilephospho-histoneH3expressionwasmarkedlydecreased(P<0．05)inneuroepitheliumfromNTDembryos．Conclusion：MTXalteredthegeneexpressionatearlyneumlation，especiallythediferentialexpressionofapoptosis—andproliferation-relatedgene

8、s，whichmaybeacriticalmechanismintheoccurrenceofNTDscausedbyfo