基于新型衍生化方法和二维液相色谱-质谱的全氟辛烷磺酸脂质组学研究

Perfluorooctane sulfonate (PFOS) was listed as persistent organic pollutants in 2009, attracting attention from domestic and overseas. PFOS has been reported to induce abnormal expressions of peroxisome proliferator activated receptors (PPARs) and cause multiple toxic effects. Although PPARs were believed to play crucial roles in lipid metabolism, the PFOS-dependent lipid variations and toxic mechanisms still remain poorly understood. This study therefore aimed to establish a novel lipidomics strategy based on a new derivatization method and two-dimensional liquid chromatography-mass spectrometry technology, and utilize the strategy in the exploration of the PFOS toxic mechanisms. To begin with, the novel derivatization reagent and method would be developed to increase the detection sensitivity of free fatty acids which have encountered severe ionization problems with liquid chromatography-mass spectrometry detection. Then the lipidomics analysis would be conducted to identify the novel lipid biomarkers and dysregulated lipid metabolic pathways perturbed by PFOS. Finally, high-throughput RNA sequencing and biological experiments would be used to confirm the lipidomics-based results and construct the lipid signaling pathways, which may provide new understandings for the therapy of PFOS-related diseases.

全氟辛烷磺酸（PFOS）是一类新型持久性有机污染物，会诱导动物体内过氧化物酶体增殖物激活受体（PPAR）的异常表达，从而引起多种毒性作用。研究表明，PPAR在脂质代谢中具有至关重要的作用，但PFOS引起的动物体内脂质类化合物代谢水平的变化仍然未知，脂质相关的PFOS毒性机制仍然是一个亟待解决的科学问题。本项目拟建立基于游离脂肪酸新型衍生化方法和二维液相色谱-质谱的脂质类化合物分析方法，从脂质组学的角度对PFOS的毒性机制进行研究。首先针对游离脂肪酸在液相色谱-质谱分析中离子化效率低的问题，开发新型衍生化试剂和方法，以提高检测灵敏度，进而提高脂质组学方法的覆盖度；随后针对PFOS暴露小鼠开展脂质组学分析，系统探究PFOS导致的脂质代谢异常；最后通过RNA测序等手段验证关键代谢通路，构建脂质相关信号传导路径，为全面了解PFOS的毒性机制，进而针对性治疗PFOS导致的疾病提供科学思路和基础数据。

全氟辛烷磺酸（PFOS）是一类新型持久性有机污染物，会诱导动物体内过氧化物酶体增殖物激活受体（PPAR）的异常表达，从而引起多种毒性作用。研究表明，PPAR在脂质代谢中具有至关重要的作用，但PFOS引起的动物体内脂质类化合物代谢水平的变化仍然未知，PFOS毒性机制仍然是一个亟待解决的科学问题。本项目首先针对游离脂肪酸在液相色谱-质谱（LC-MS）分析中离子化效率低的问题，开发了新型衍生化试剂2’-羰基-哌嗪罗丹明B（CPR），构建了游离脂肪酸的新型衍生化方法，并在此基础上建立了基于新型衍生化方法和二维LC-MS的脂质组学分析方法；随后针对PFOS暴露大鼠开展脂质组学分析，探索了PFOS导致的脂质代谢异常，揭示了甘油磷脂代谢、胆碱代谢、逆向内源性大麻素信号等异常代谢通路；最后通过RNA测序、代谢组学等手段验证关键代谢通路，构建了PFOS暴露导致的相关信号传导路径，为全面了解PFOS的肝毒性、免疫毒性、神经毒性等毒性机制，进而针对性治疗PFOS导致的疾病提供了科学思路和基础数据。