利用SWATH-MS分析小鼠小腸中塑膠微粒毒性的差異蛋白質體學

Abstract

由於人類大量釋放塑膠垃圾於環境中，這些垃圾會被逐一分解成顆粒更小的塑膠微粒，並可能經由食物鏈回到人體，產生生物放大作用的影響。先前文獻指出，塑膠微粒會在哺乳動物的胃腸道、肝臟、腎臟和大腦中積累。本研究利用SWATH這項無標記定量的質譜技術，分析小鼠小腸中的聚苯乙烯微粒實驗組和控制組的差異蛋白質。先將胜肽進行第一維的鹼性逆向層析分離後，再送到後續奈米液相層析串聯質譜分析(nanoLC-MS/MS)。本研究不僅成功地鑑定到3666蛋白質及其31501的胜肽片段，同時也用SWATH方法定量出2933個蛋白質。在實驗組和控制組中有473個蛋白質表現量具有顯著差異。藉由GO和Metascape分析，可以看到這些具有差異性的蛋白質主要和細胞凋亡、發炎反應、代謝混亂等途徑相關。而這些蛋白質將來也有望成為因聚苯乙烯微粒導致腸道損傷的潛在生物標記物，同時也為塑膠微粒致使腸道受損的毒理性提供新的見解。The massive release of plastic waste into the environment by humans can lead to the breakdown of these materials into smaller particles known as microplastics (MPs). These MPs can then be transferred to humans via the food chain, potentially causing biomagnification effects. Previous studies have shown that microplastics can accumulate in the gastrointestinal tract, liver, kidneys, and brain of mammals. This study used SWATH, an untargeted quantitative mass spectrometry technique, to analyze the differential proteins in the small intestine of mice exposed to polystyrene microplastics compared to a control group. The digested peptides were separated by first-dimension alkaline reversed-phase chromatography and then analyzed by nanoLC-MS/MS. Our study has maximized the size of ion database for protein identification and quantification by combining 3 fractionations and were able to identify 3666 proteins and 31501 peptides. Additionally, 2933 proteins were quantified using the SWATH method. A total of 473 proteins exhibited significant differences in expression between the experimental and control groups. Metascape analyses revealed that these differentially expressed proteins were primarily involved in cell apoptosis, inflammatory responses, , and metabolic disturbances. The study also identified several potential biomarkers for PS-MPs-induced intestinal damage. These biomarkers could be used to develop new methods for detecting and monitoring PS-MPs exposure and its health effects.