利用SWATH及iTRAQ技術進行小鼠腎臟中塑膠微粒毒性之差異蛋白質體學分析

Abstract

根據聯合國的數據，每年有超過800萬噸的塑料垃圾被丟棄於海洋。這些塑膠垃圾會逐漸分解成塑膠微粒，最終可能積聚在人類食物鏈中。已有相關研究指出，塑膠微粒會積累在肝臟、腎臟和腸道等器官中。本研究使用了SWATH和iTRAQ兩種差異蛋白質技術，分析塑膠微粒投餵組和對照組的蛋白質體學。為了減少樣品的複雜度，腎臟蛋白質樣品經胰蛋白酶水解過後的胜肽，先經鹼性逆相層析法分離，接著進行奈米液相層析串聯質譜分析(nanoLC-MS/MS)。SWATH和iTRAQ在相對定量方面具有相似的趨勢，並分別成功定量了1984和2162個蛋白質，塑膠微粒投餵組和對照組之間有顯著差異的蛋白質有489個。透過GO及Metascape分析，發現這些差異表達的蛋白質主要與細胞過程、代謝過程和生物調節途徑有關。未來這些蛋白質有望成為塑膠微粒引起的腎損傷的潛在生物標記物，這項研究為塑膠微粒誘導腎損傷的機制提供新的見解。According to the United Nations, more than 8 million tons of plastic waste are discarded in ocean every year. It will be physically or chemically decomposed into microplastics, which may eventually accumulate in the human food chain. It has been reported that it will accumulate in organs such as liver, kidney and intestine. Two differential proteomic technologies, SWATH and iTRAQ, were used to investigate protein profiles in microplastic-treated groups and control group in this study. To reduce sample complexity, the proteins from kidney samples were tryptic digested and fractionated by alkaline reverse phase chromatography, followed by nano liquid chromatography tandem mass spectrometry (nanoLC-MS/MS). The SWATH and the iTRAQ gave similar analytical performance. Overall, 1984 and 2162 proteins were successfully quantified by the SWATH and the iTRAQ, respectively. Microplastic-treated groups showed significant differences in 489 proteins treated and control groups. Based on the Gene Ontology and Metascape analysis, these differential expressed proteins were found to be mainly related to cellular process, metabolic process, and biological regulation pathways. These proteins are expected to serve as potential biomarkers of microplastic-induced kidney injury.