秀丽隐杆线虫生殖系统调控寿命的机制研究

Revealing the cause for aging and anti-aging mechanism will help to answer how to enter aging population healthily. The insulin/ins signaling pathway and dietary restriction pathway are the main aging modulation pathway related with nutrition regulation and have an important role in reproduction development and reproductive aging. The reproductive system of Caenorhabditis elegans includes somatic gonad and germline. Somatic gonad and germline send out unknown anti and pro-aging signals, respectively. In our previous study, we have obtained the differential expression proteins and metabolites between germline-less mutant of glp-1 and germline stem cell tumorogenesis mutant of gld-2 through proteomic and metabolomics investigation. In this project, we will continue to investigate the mechanism of reproduction system on aging regulation in a systemic biology approach, such as proteomics, metabolomics and transcriptome study. These researches would provide the protein and gene expression and metabolite profile related to aging. We will using RT-PCR, Western blot to confirm the genes discovered to relate with aging. We will also confirm those metabolites related with aging phenotypes using metabolite standard. Further investigation of the discovered underlying molecular pathways on regulation of aging would be performed rigorously using RNAi or mutant strains. This study will provide new information on the mechanism of aging and help to solve the aging related problems.

阐明衰老的根本原因和对抗衰老的长寿机制，有助于解决如何健康的进入老年群体等问题。调控衰老的胰岛素和节食相关信号通路可归结为营养相关信号通路，并调控着生殖发育和生殖寿命，而生殖系统是另一个主要的衰老调控者。秀丽线虫生殖系统分为生殖干细胞系和体细胞系，各自通过未知分子对机体发出缩短或延长寿命的信号。我们前期工作已通过蛋白质组和代谢组初步获得与衰老相关的差异表达蛋白和代谢产物。本项目将继续用系统生物学的方法，对生殖干细胞缺失突变体glp-1和生殖干细胞增生突变体gld-2的进行蛋白质组、代谢组和转录组学研究，比较其基因表达和代谢产物差异，并通过RT-PCR、蛋白免疫印迹和代谢产物标准品进行验证。然后通过RNAi和相关突变体的遗传分析，挖掘生殖干细胞和体细胞各自特异的基因表达和发出的寿命调控信号分子，获得生殖系统调控衰老的较为全面的认识，为解决我国日趋严峻的老龄化问题提供理论基础。

阐明衰老的根本原因和对抗衰老的长寿机制，有助于解决如何健康的进入老年群体等问题。调控衰老的胰岛素和节食相关信号通路可归结为营养相关信号通路，并调控着生殖发育和生殖寿命，而生殖系统是另一个主要的衰老调控者。秀丽线虫生殖系统分为生殖干细胞系和体细胞系，各自通过未知分子对机体发出缩短或延长寿命的信号。本项目利用系统生物学的方法，对生殖干细胞缺失突变体glp-1和生进行蛋白质组和代谢组进行研究，比较其基因表达和代谢产物差异，并通过RT-PCR、蛋白免疫印迹和代谢产物标准品进行验证。然后通过本项目的研究，挖掘生殖干细胞特异的基因表达和发出的寿命调控信号分子，获得了生殖系统调控衰老的较为全面的认识，为解决我国日趋严峻的老龄化问题提供了理论基础。本研究结果表明：.1. glp-1介导的生殖系统通过影响蛋白质组和代谢组调控寿命。线虫glp-1 功能缺失突变体（减少 germline干细胞）会导致寿命的延长，而这种寿命的延长在某种意义上是通过影响代谢水平和蛋白质水平进而调控了寿命。.2. glp-1功能缺失的线虫中生殖细胞丢失导致参与蛋白质合成和分解的蛋白质丰度降低，参与解毒和免疫应答的蛋白质丰度增加，表明蛋白质合成和代谢减弱，而解毒和免疫应答增加。蛋白质丰度的改变可能导致生理适应，有助于延长生殖系缺乏的寿命。.3. glp-1功能缺失线虫通过调控许多衰老相关水相小分子代谢实现寿命的延长，主要包括影响三羧酸循环、甘油磷脂代谢、嘧啶代谢、淀粉和蔗糖代谢以及嘌呤代谢产物，以及通过调控许多衰老相关脂质分子代谢来延长寿命，如PC 44:2、 TG 48:4、TG 50:1等。.4. glp-1功能缺失线虫寿命延长仅仅部分依赖于DAF-16转录因子，DAF-16仅介导了部分glp-1功能缺失线虫衰老相关的代谢改变， 如三羧酸循环、嘧啶代谢、 DG 34:3、 TG 49:2和TG 51:3等。