维甲酸诱导神经管畸形小鼠组蛋白H2BK120ub1泛素化修饰及相关基因表达变化  

Ubiquitination modification of histone H2BK120ub1 and expression of related genes in retinoic acid-induced neural tube defects

作　　者：王珊[1] 成翕悦[2] 裴培[1] 刘帆 占小俊[4] WANG Shan;CHENG Xi Yue;PEI Pei;LIU Fan;ZHAN Xiao Jun(Beijing Municipal Key Laboratory of Child Development and Nutriomics,Capital institute of pediatrics,Beijing 100020,China)

机构地区：[1]首都儿科研究所儿童发育与营养重点实验室,北京100020 [2]河北医科大学第三医院全科医学科,石家庄050000 [3]北京协和医学院研究生院,100005 [4]首都儿科研究所耳鼻喉头颈外科,北京100020

出　　处：《中国生育健康杂志》2022年第5期444-449,共6页Chinese Journal of Reproductive Health

基　　金：国家自然基金面上项目(82071690);首都儿科研究所所级基金(FX-2020-05,CXYJ-2-21-09);北京市医管中心“使命”人才培养计划项目-耳鼻咽喉科学(No.SML20190601)。

摘　　要：目的 利用维甲酸(RA)诱导C57BL/6J小鼠神经管畸形(NTDs),探讨RA对NTDs鼠胚脑、脊髓组织组蛋白H2BK120单泛素化修饰水平及神经管发育基因表达水平变化。方法 孕鼠随机对照(Con)组和RA构建的NTDs组。实验组孕鼠7.5 d灌胃RA 25 mg/kg;对照组孕鼠灌胃相同体积的香油。孕10.5 d通过体视显微镜观察胎鼠畸形及神经管发育。采用Elisa方法检测NTDs小鼠胚脑、脊组织维生素A、维甲酸浓度的变化,采用Western blot检测NTDs小鼠胚脑、髓组织H2BK120ub1蛋白的表达量变化;采用RT-PCR检测NTDs小鼠胚脑、脊组织神经管发育基因Pax6、Nestin表达变化;采用ChIP-qPCR检测NTDs小鼠胚脑、脊组织神经管发育基因Pax6、Nestin与H2BK120ub1结合变化。结果 RA组诱导NTDs胚鼠脑、脊髓组织:维甲酸及维生素A浓度相较于对照组明显增高(P<0.05);H2BK120ub1蛋白表达水平明显高于对照组(P<0.05);神经管发育基因Pax6、Nestin表达水平明显高于对照组(P<0.05);神经管发育基因Pax6、Nestin与H2BK120ub1结合显著增强(P<0.05)。结论 神经管发育基因Pax6、Nestin和H2BK120单泛素化在胚胎中的异常表达与维甲酸致神经管畸形存在相关性。Objective To induce neural tube defects(NTDs) in C57 BL/6 J mice by retinoic acid(RA) and to investigate the effects of RA on the changes of histone H2 BK120 monoubiquitination and neural tube development gene expression levels in the brain and spinal cord tissues of NTDs mice. Methods Pregnant mice were randomized the control group and the RA-treatment group. NTDs mouse embryos were induced by gavage with 25 mg/kg(body weight) of RA on E7.5(RA is dissolved in sesame oil). On E10.5, pregnant mice were euthanized by cervical dislocation. Elisa detected levels of retinoid and vitamin A. Western blot and immunohistochemical staining to detect the expression of H2 BK120 ub1. Real-time quantitative polymerase chain reaction(RT-qPCR) is used to detect the expression of Pax 6, nestin and h2 bk120 ub1. Results On E10.5, the concentration of retinoid and vitamin A were higher in RA-induced cranial neural tissue than controls(P<0.05). The expression of H2 BK120 ub1 protein was significantly higher in RA-induced cranial neural tissue than controls(P<0.05). The mRNA levels of neural tube gene, Pax6 and Nestin were significantly higher in NTD embryos compared to those in controls(P<0.05). The binding of Pax6 and Nestin genes with H2 BK120 ub1 was significantly enhanceds(P<0.05). Conclusion Abnormal expression of single ubiquitination of neural tube development genes Pax6, nestin and h2 bk120 in embryos was related to neural tube malformations caused by retinoic acid.

关 键 词：维甲酸 神经管畸形 H2BK120单泛素化表达 

分 类 号：R17[医药卫生—妇幼卫生保健] R71[医药卫生—公共卫生与预防医学]