Ates Eda, My Ong Hien Thi, Yu Seung-Min, Kim Ji-Hoon, Kang Min-Jung
Center for Advanced Biomolecular Recognition, Biomedical Research Institute, Korea Institute of Science and Technology, Seoul, Republic of Korea; Division of Bio-Medical Science & Technology, KIST School, University of Science and Technology, Seoul, Republic of Korea.
Center for Advanced Biomolecular Recognition, Biomedical Research Institute, Korea Institute of Science and Technology, Seoul, Republic of Korea; College of Medicine, Seoul National University, Seoul, Republic of Korea.
Mol Cell Proteomics. 2025 Mar;24(3):100921. doi: 10.1016/j.mcpro.2025.100921. Epub 2025 Jan 31.
Nonalcoholic fatty liver disease is a hepatic condition characterized by excessive fat accumulation in the liver with advanced stage nonalcoholic steatohepatitis (NASH), potentially leading to liver fibrosis, cirrhosis, and cancer. Currently, the identification and classification of NASH require invasive liver biopsy, which has certain limitations. Mass spectrometry-based proteomics can detect crucial proteins and pathways implicated in NASH development and progression. We collected the liver and serum samples from choline-deficient, L-amino acid-defined high-fat diet fed NASH C57BL/6J mice and human serum samples to examine proteomic alterations and identify early biomarkers for NASH diagnosis. In-depth targeted multiple reaction monitoring scanning and immunoblotting assays were used to verify the biomarker candidates from mouse liver and serum samples, and enzyme-linked immunosorbent assay (ELISA) was employed to analyze human serum samples. The multiple reaction monitoring analysis of NASH liver revealed 50 proteins with altered expression (21 upregulated and 29 downregulated) that are involved in biological processes such as detoxification, fibrosis, inflammation, and fatty acid metabolism. Ingenuity pathway analysis identified impaired protein synthesis, cellular stress and defense, cellular processes and communication, and metabolism in NASH mouse liver. Immunoblotting analysis confirmed that the expression of proteins associated with fatty acid metabolism (Aldo B and Fasn) and urea cycle (Arg1, Cps1, and Otc) was altered in the mouse liver and serum. Further analysis on human serum samples using ELISA confirmed the increased expression of multiple proteins, including Aldo B, Asl, and Lgals3, demonstrating values of 0.917, 0.979, and 0.965 of area under the curve in NASH diagnosis. These findings offer valuable insights into the molecular mechanisms of NASH and possible diagnostic biomarkers for early detection.
非酒精性脂肪性肝病是一种肝脏疾病，其特征是肝脏中脂肪过度积累，晚期可发展为非酒精性脂肪性肝炎（NASH），可能导致肝纤维化、肝硬化和癌症。目前，NASH的识别和分类需要进行侵入性肝脏活检，这存在一定局限性。基于质谱的蛋白质组学可以检测与NASH发生和发展相关的关键蛋白质和信号通路。我们收集了胆碱缺乏、L-氨基酸定义的高脂饮食喂养的NASH C57BL/6J小鼠的肝脏和血清样本以及人类血清样本，以检查蛋白质组学变化并鉴定用于NASH诊断的早期生物标志物。采用深度靶向多反应监测扫描和免疫印迹分析来验证小鼠肝脏和血清样本中的候选生物标志物，并使用酶联免疫吸附测定（ELISA）分析人类血清样本。对NASH肝脏的多反应监测分析显示，有50种蛋白质表达发生改变（21种上调，29种下调），这些蛋白质参与解毒、纤维化、炎症和脂肪酸代谢等生物学过程。 Ingenuity通路分析确定NASH小鼠肝脏中蛋白质合成、细胞应激和防御、细胞过程和通讯以及代谢受损。免疫印迹分析证实，与脂肪酸代谢（Aldo B和Fasn）和尿素循环（Arg1、Cps1和Otc）相关的蛋白质在小鼠肝脏和血清中的表达发生了改变。使用ELISA对人类血清样本进行的进一步分析证实，包括Aldo B、Asl和Lgals3在内的多种蛋白质表达增加，在NASH诊断中曲线下面积值分别为0.917、0.979和0.965。这些发现为NASH的分子机制以及早期检测的可能诊断生物标志物提供了有价值的见解。
