摘要
神经管畸形(NTDs)的病因与防治是出生缺陷领域研究的重点,叶酸可以预防神经管畸形但其机制不明。本文借助低叶酸细胞模型和低叶酸NTDs小鼠模型通过染色质免疫共沉淀、Cut&Tag等技术,探讨了组蛋白去甲基化酶lysine demethylase 5A(KDM5A)及其调控的下游组蛋白H3K4me3修饰在叶酸缺乏导致的NTDs发生中的潜在分子机制。结果显示,低叶酸的细胞模型中,qRT-PCR、Western印迹结果显示,KDM5A分子表达明显下降(P<0.05)。作为组蛋白H3K4me3调控的上游关键酶,进一步通过染色质免疫共沉淀ChIP、ChIP-qPCR实验证实,叶酸缺乏下组蛋白H3K4me3在神经发育基因Axin 2和Atoh 1基因启动子区富集增加(P<0.05)。通过构建KDM5A基因敲除细胞模型,借助Cut&Tag试验证实,KDM5A基因敲除后H3K4me3主要富集在神经发育基因上。最后在低叶酸导致的NTDs小鼠模型的脑组织中,RT-qPCR、Western印迹以及ChIP-qPCR实验显示,E9.5 d的NTDs胎鼠脑组织中KDM5A表达下降(P<0.05),Axin2、Atoh1表达升高(P<0.05),Axin2、Atoh 1基因启动子区的H3K4me3富集增多(P<0.05)。综上所述,KDM5A蛋白在叶酸缺乏导致的NTDs中发挥重要作用,其可通过调控下游H3K4me3进而调控神经发育靶基因Axin2、Atoh 1异常表达,介导NTDs的发生。本研究从叶酸缺乏介导KDM5A调控组蛋白修饰来探讨NTDs的发病机制,为降低出生缺陷,促进生殖健康提供依据。
Neural tube defects(NTDs)represent a critical area of study within congenital anomalies,with folate known for its preventative role.However,the mechanisms underlying its protective effects remain largely unknown.This study investigates the potential molecular mechanisms involving lysine demethylase 5A(KDM5A)and its subsequent alteration of histone H3K4me3 in the development of NTDs under folate deficiency.We used chromatin immunoprecipitation along with Cut&Tag to examine the function of KDM5A in a low-folate cell model and a mouse model of folate-deficient NTDs.Quantitative reverse transcription PCR(qRT-PCR)and Western blot analyses demonstrated a marked reduction in KDM5A expression in the low-folate cell model(P<0.05).In addition,chromatin immunoprecipitation(ChIP)followed by quantitative PCR(ChIP-qPCR)analysis confirmed the increased accumulation of histone H3K4me3 in the promoter regions of important neurodevelopmental genes,Axin2 and Atoh1,with folate deficiency(P<0.05).By creating a KDM5A-knockout cell model,Cut&Tag experiments was used to confirm the preferential enrichment of H3K4me3 on neurodevelopmental genes.In the brains of folate-deficient NTDs models,decreased expression of KDM5A and upregulated Axin2 and Atoh1 expression were observed,along with increased H3K4me3 enrichment at respective gene promoters(P<0.05).Collectively,these findings highlight the important function of KDM5A in NTDs with folate deficiency.KDM5A affects the expression of genes related to neurodevelopment by modifying H3K4me3 downstream.This study enhances our comprehension of the development of NTDs by examining the impact of disrupted folate metabolism and abnormal regulation of histone modification by KDM5A.It provides valuable insights into potential treatments for reducing birth defects and improving reproductive health.
作者
李建婷
解琪
谷小龙
曹志华
彭志伟
赵虹
刘志贞
解军
LI Jian-Ting;XIE Qi;GU Xiao-Long;CAO Zhi-Hua;PENG Zhi-Wei;ZHAO Hong;LIU Zhi-Zhen;XIE Jun(Shanxi Key Laboratory of Birth Defect and Cell Regeneration,Department of Biochemistry and Molecular Biology,School of Basic Medicine,Shanxi Medical University,Taiyuan 030001,China;Key Laboratory of Coal Environmental Pathogenicity and Prevention(Ministry of Education,China),Taiyuan 030001,China)
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2024年第5期629-637,共9页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家自然科学基金(No.U23A20420,No.82201841)
山西省基础研究计划(No.202203021212369)
山西医科大学博士启动基金项目(No.SD2209,No.XD2116)资助。
