糖尿病诱发神经管畸形的组蛋白乙酰化修饰谱  

Histone acetylation profile of diabetes induced neural tube defects

作　　者：白宝玲[1] 万春蕾 李丹 周珍珍 张勤[1] BAI Baoling;WAN Chunlei;LI Dan;ZHANG Qin(Department of Biochemical and Immune Research,Capital Institute of Pediatrics,Beijing 100020,China)

机构地区：[1]北京首都儿科研究所生化免疫室,北京100020 [2]通州区妇幼保健院儿科

出　　处：《中国糖尿病杂志》2020年第10期766-773,共8页Chinese Journal of Diabetes

基　　金：国家自然科学基金(81901167)。

摘　　要：目的探讨母亲DM诱发胎儿神经管畸形(NTDs)的组蛋白乙酰化修饰机制。方法构建高糖处理小鼠神经干细胞(NE-4C)模型、T1DM雌鼠诱发NTDs胎鼠模型及人类母亲高血糖相关NTDs胚胎脑组织,建立葡萄糖浓度5、25 mmol/L的NE-4C细胞模型组;雌鼠分为正常对照(NC)组(n=40)和STZ构建的TIDM组(n=80);人类样本分为对照(Con)组和NTDs组。Western blot联合Orbitrap QE-HF质谱仪定性分析组蛋白乙酰化修饰谱,定量检测组蛋白特定位点乙酰化修饰水平。结果25 mmol/L组相比5 mmol/L组,TIDM组相比NC组,NTDs相比Con组,组蛋白赖氨酸位点整体乙酰化修饰位点均增加,其中组蛋白H4K5/K8/K12/K16ac修饰在体外体内模型均保守性修饰,且25 mmol/L组H4K5/K8/K12/K16ac修饰水平是5 mmol/L组的2.2倍(P<0.01)。结论组蛋白乙酰化修饰异常增加可能参与调控母亲DM诱发NTDs。Objective To investigate the mechanism of histone acetylation in fetal neural tube defects(NTDs)induced by maternal diabetes mellitus(DM).Methods High glucose treated mouse neural stem cell(NE-4 C)model,diabetic female induced NTDs fetal mouse model and human maternal hypergly-cemia related NTDs embryonic brain tissue were constructed.NE-4 C Cells were divided into 5 mmol/L and25 mmol/L group.Female mice were divided into normal contro(lNC)group(n=40)and STX induced T1 DM group(n=80).Human samples were divided into normal control group(Con group)and NTDs group.The histone acetylation modification profile was qualitatively analyzed by Western blot combined with Orbitrap QE-HF mass spectrometry,and the acetylation modification level of histone specific sites was quan-titatively detected.Results The whole acetylation sites of histone lysine site were increased in 25 mmol/L group compared with 5 mmol/L group,TIDM group compared with NC group,and NTDs group compared with Con group.The histone H4 K5/K8/K12/K16 ac modification was conservative in vitro and in vivo model,and the modification level of H4 K5/K8/K12/K16 ac was 2.2 times higher in 25 mmol/L group than in 5 mmol/L group(P<0.01).Conclusion Abnormal increase of histone acetylation may be involved in the regulation of NTDs induced by maternal diabetes.

关 键 词：Orbitrap QE-HF质谱仪 平行反应监测 母亲糖尿病 神经管畸形 组蛋白乙酰化修饰 

分 类 号：R587.2[医药卫生—内分泌]