甲氨蝶呤对早期神经胚基因表达的影响  

Alteration of Gene Expression by Methotrexate Exposure at Early Neurulation

作　　者：季成[1] 王秀伟[2] 王建华[2] 官臻[2] 朱智强[2] 谢秋[2] 张霆[2] 牛勃[1,2] 

机构地区：[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

出　　处：《现代生物医学进展》2014年第26期5058-5062,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81070491)

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(4.5 mg/kg体重)干预孕鼠,通过NimbleGene表达谱芯片、Real time-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(fold change>1.5,P<0.05),3个增殖相关基因(Ptch1,Pla2g4a,Foxg1)均表现为下调(fold change<0.67,P<0.05);NTDs胚胎神经上皮Caspase-3表达显著升高(P<0.05),phospho-histone H3(pH3)表达显著降低(P<0.05)。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。Objective： We used methotrexate （MTX） intervention in pregnant C57BL/6J mice to investigate the alteration of gene expression at early neurulation. Methods： The differential gene expressions were studied by Microarray, RT-PCR and Immun ohistochemical assays in NTD embryos induced by MTX （4.5 mg/kg body weight. Results： Results showed that 32.1% of NTDs was developed by the treatment of MTX. Microarray indicated that 166 genes were significantly different between control and NTD mice, including 4 apoptosis-related genes （Endog, Trp53, Casp3, Bax） and 3 proliferation-related genes （Ptchl, Pla2g4a, Foxgl）. Levels of Endog, Trp53, Casp3, Bax （fold change 〉1.5） were up-regulated but Ptchl, Pla2g4a, Foxgl （fold change 〈0.67） were down-regulated （P〈0.05）. Expression of caspase-3 was significantly enhanced （P〈0.05） while phospho-histone H3 expression was markedly decreased （P〈0.05） in neuroepithelium from NTD embryos. Conclusion： MTX altered the gene expression at early neumlation, especially the differential expression of apoptosis-and proliferation-related genes, which may be a critical mechanism in the occurrence of NTDs caused by folate deficiency.

关 键 词：神经管畸形 甲氨蝶呤 凋亡 增殖 

分 类 号：Q95-3[生物学—动物学] R722[医药卫生—儿科]