乙硫氨酸引起小鼠神经管畸形的机制探讨  

Mechanism of neural tube defects caused by ethionine in mice

作　　者：王文卓 李美宁[1] 张丽[2] 李丹丹 赵晓荣 牛玉虎[1] 牛勃[1,3] WANG Wenzhuo;LI Meining;ZHANG Li;LI Dandan;ZHAO Xiaorong;NIU Yuhu;NIU Bo(不详;Department of Biochemistry and Molecular Biology,Shanxi Medical University,Taiyuan 030001,Shanxi Province,China)

机构地区：[1]山西医科大学生物化学与分子生物学教研室,山西太原030001 [2]山西医科大学第一医院肝胆外科及肝脏移植中心,山西太原030001 [3]首都儿科研究所儿童发育营养组学北京市重点实验室,北京10020

出　　处：《中国生物制品学杂志》2024年第6期666-671,共6页Chinese Journal of Biologicals

基　　金：山西省自然科学基金(201801D121313)

摘　　要：目的利用甲硫氨酸结构类似物——乙硫氨酸制备小鼠神经管畸形(neural tube defects,NTDs)模型,并探讨NTDs发生对神经细胞增殖、凋亡及迁移的影响。方法建立小鼠NTDs胚胎模型,C57BL/6J小鼠有孕至7.5 d(E7.5)时,经腹腔注射500 mg/kg乙硫氨酸,对照组注射等剂量生理盐水,E11.5时取小鼠胚胎,体式显微镜下观察胚胎形态;ELISA检测孕鼠血浆中S-腺苷同型半胱氨酸(S-adenosylhomocysteine,SAH)、S-腺苷甲硫氨酸(S-adenosylmethionine,SAM)水平;Western blot及RT-PCR法检测胚胎脑组织迁移蛋白E-cadherin和N-cadherin的表达及基因mRNA的转录,Western blot法检测细胞增殖蛋白PCNA、凋亡蛋白cleaved-Caspase3及Wnt/β-catenin信号通路标志蛋白β-catenin、TCF4、C-myc的表达。结果乙硫氨酸干预后,孕鼠体内SAM含量减少,SAH含量增加,SAM/SAH降低。与对照组比较,NTDs组小鼠E-cadherin、cleaved-Caspase3高表达,细胞凋亡增多;N-cadherin、PCNA低表达,细胞增殖、迁移减少;且Wnt/β-catenin信号通路标志蛋白β-catenin、TCF4、C-myc均低表达,表明通路被抑制。结论乙硫氨酸可能通过抑制Wnt/β-catenin信号通路及神经细胞的增殖与迁移,促进细胞凋亡,进而引起NTDs。Objective To prepare a neural tube defects(NTDs)model in mice with the structural analogue of methionineethionine,and investigate the effects of NTDs on the proliferation,apoptosis and migration of nerve cells.Methods Mouse NTDs embryo model was established.The C57BL/6J mice were injected intraperitoneally with 500 mg/kg ethionine after gestation to 7.5 d(E7.5),while the mice in control group were injected with the same dose of normal saline.Mouse embryos were taken at E11.5 and observed under stereomicroscope;The levels of S-adenosylhomocysteine(SAH)and Sadenosylmethionine(SAM)in plasma of pregnant mice were detected by ELISA;Western blot and RT-PCR were used to detect the expression of migration proteins E-cadherin and N-cadherin and the mRNA transcription in embryonic brain tissue.The expression of proliferation protein PCNA and apoptosis protein cleaved-Caspase3,and Wnt/β-catenin signaling pathway marker proteinsβ-catenin,TCF4 and C-myc were detected by Western blot.Results After the intervention of ethionine,the SAM content in pregnant mice decreased,the SAH content increased,and the SAM/SAH ratio decreased.Compared with the control group,the E-cadherin and cleaved-Caspase 3 were highly expressed and the apoptosis increased in NTDs group;The expression of N-cadherin and PCNA decreased,and the cell proliferation and migration decreased;The low expression of Wnt/β-catenin signaling pathway marker proteinsβ-catenin,TCF4 and C-myc indicated that the pathway was inhibited.Conclusion Ethionine may cause NTDs by promoting cell apoptosis via inhibiting Wnt/β-catenin signaling pathway and the proliferation and migration of nerve cells.

关 键 词：神经管畸形 乙硫氨酸 迁移 WNT/Β-CATENIN信号通路 

分 类 号：R321[医药卫生—人体解剖和组织胚胎学]