微囊藻毒素-LR致大鼠睾丸生殖细胞凋亡的表观遗传调控机制

项目名称： 微囊藻毒素-LR致大鼠睾丸生殖细胞凋亡的表观遗传调控机制

项目编号： No.81472948

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 张慧珍

作者单位： 郑州大学

项目金额： 70万元

中文摘要： 微囊藻毒素LR（MC-LR）是污染严重、人群接触广、危害大的微囊藻毒素异构体，其生殖毒性受到了广泛关注和重视。本课题组研究表明MC-LR可改变凋亡相关基因表达，诱导细胞凋亡，但其分子机制尚不明确。表观遗传改变调控凋亡相关基因的表达，可能在MC-LR致睾丸生殖细胞凋亡中起着调控作用。为此，本项目以MC-LR为受试物，以体外共培养的睾丸支持细胞和生精细胞为细胞模型，以青春前期、青春期SD大鼠为动物模型，应用染色质免疫共沉淀联合芯片技术、激光扫描共聚焦显微技术等方法，从动物、细胞和分子水平检测MC-LR通过线粒体介导的caspase依赖通路诱导的睾丸生殖细胞凋亡，分析H3K4me3、SIRT1和组蛋白乙酰化在MC-LR致睾丸生殖细胞凋亡中的调控作用，从表观遗传学角度探索MC-LR致睾丸生殖细胞凋亡的分子机制，为全面深入认识和理解MC-LR的生殖毒性机制提供新思路，为保护人群生殖健康提供理论基础。

中文关键词： 微囊藻毒素-LR；生殖毒性；大鼠睾丸；凋亡；表观遗传

英文摘要： Microcystin-LR(MC-LR), a naturally occurring toxin produced by cyanobacteria, is a microcystins isomer and considered the most abundant and toxic compound. Therefore, many studies have been focused on reproductive toxicity of MC-LR in mammals including humans. Our previous results showed that the expression of apoptosis-related genes can be regulated by MC-LR, indicating that apoptosis plays an important role in the reproductive toxicity of MC-LR. Up to now, molecular mechanism of reproductive toxicity of MC-LR is still unclear. It was reported that epigenetic modification can regulate the expression of apoptosis related gene, suggesting that epigenetic modification may be involved in the cell apoptosis induced by MC-LR. In this project, MC-LR will be used as test sample, the co-culture of sperm cells with testis sertoli cells will be used as cell model, and prepubertal and pubertal rats will be used as animal model. The testicular damage caused by MC-LR in rats will be explored with Chromatin Immunoprecipitation-chip (ChIP-chip), Laser scanning confocal microscopy, and so on. The cell apoptosis induced by MC-LR through mitochondrial mediated caspase dependent pathway will be surveyed at the anmial, cell and molecular level. The regulation function of H3K4me3, SIRT1 and acetyl-Histone in cell apoptosis will also be anaylazed, revealing molecular mechanism of testicular germ cell apoptosis caused by MC-LR at the epigenetic level. The study will provide a new train of idea for comprehensive understanding reproductive toxicity of MC-LR, and lay the foundation for preventing people health from algal toxins.

英文关键词： microcystin-LR;reproductive toxicity;rat testis;apoptosis;epigenetic