蛋白乙酰化介导镉离子诱导小鼠生殖细胞损伤毒理机制研究

重金属是人类生殖疾病及动物繁殖障碍的关键诱因之一，然而相关生殖毒性的分子机制尚未明确。申请人前期研究发现，镉离子能抑制小鼠睾丸生殖细胞蛋白乙酰化水平及影响Tnps、Prms替换组蛋白过程，预示蛋白乙酰化介导重金属离子诱导小鼠生殖细胞损伤。为阐明镉离子调控蛋白乙酰化介导生殖细胞损伤的分子机制，本项目在前期研究基础上，利用细胞转录组学、代谢组学、修饰肽段富集及Chip-seq技术，系统研究Cd2+暴露下睾丸生殖细胞蛋白乙酰化修饰、差异表达基因及TCA代谢产物变化，探究Cd2+抑制生殖细胞蛋白乙酰化修饰的作用机制，重点解析微管蛋白乙酰化被抑制后对减数分裂的影响，阐明组蛋白H4K5、H4K12乙酰化抑制与关联基因转录表达的关联性以及Cd2+抑制组蛋白乙酰化与Tnps、Prms替换组蛋白的相互关系，旨为揭示Cd2+抑制蛋白乙酰化介导小鼠睾丸生殖细胞损伤的分子机制，为重金属毒性疾病防治提供新思路。

Heavy metal is one of the main causes of human reproductive diseases and animal reproductive disorders. Nevertheless, the relevant reproductive toxicological mechanisms are not yet clear. The applicant's previous researches demonstrated that cadmium ions can inhibit acetylation of proteins in mouse testicular spermatogenesis, and affect the replacement processes of Tnps and Prms by histones, which suggested that protein acetylation may be involved in cadmium-mediated inhibitions of mouse spermatogenesis and sperm maturation. To elucidate the molecular mechanism of spermatogenesis damage mediated by cadmium-regulated protein acetylation, based on our previous Cd2+ reproductive toxicology and acetylation proteomics research, we utilize cell transcriptomics, metabolomics, modified peptide antibody affinity enrichment technology and ChIP-seq technologies to systematically study the changes of protein acetylation during spermatogenesis, differentially expressed genes and TCA metabolites upon Cd2+ exposure. We will explore the mechanism of Cd2+-induced inhibition of protein acetylation modification during spermatogenesis, and focus on analyzing the association between H4K5 and H4K12 acetylation inhibition and transcriptional expression of related genes, and exploring the interaction between acetylation inhibition by Cd2+and the replacement of Tnps and Prms by histones. This project aim at systematically clarifying the molecular mechanism of sperm cell damage mediated by cadmium-inhibited protein acetylation in mouse. The significance of the project is to provide theoretical basis for solving the scientific problem of Cd2+-reproductive toxicology mechanism, and provide novel sights for the prevention and treatment of toxic diseases by heavy metals.