Identification of Early Proteomic Markers for Hepatic Steatosis

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• 作者：G. Meneses-Lorente ; A. Watt ; K. Salim ; S. J. Gaskell ; Nagaraja Muniappa ; Jeffrey Lawrence ; P. C. Guest
• 刊名：Chemical Research in Toxicology
• 出版年：2006
• 出版时间：August 2006
• 年：2006
• 卷：19
• 期：8
• 页码：986 - 998
• 全文大小：1021K
• 年卷期：v.19,no.8(August 2006)
• ISSN：1520-5010

文摘

The identification of biomarkers for disease state, drug efficacy, and toxicity is becoming increasinglyimportant for drug discovery and development. We have used two-dimensional differential in-gelelectrophoresis and mass spectrometry to identify proteomic markers associated with hepatocellular steatosisin rats after dosing with a compound (CDA) in preclinical development. Rats were dosed daily for up to5 days with CDA for measurement of blood biochemical parameters, histological, and proteomic analysis.Alterations in plasma glucose and liver transaminases were detected from dosing day 3 onward, andlivers showed trace levels of hepatocellular vacuolation from 6 h which increased in extent and severityover the 5 day time course. The number of significantly altered protein spots increased over the 5 daytime course, and Ingenuity Pathway Analysis showed that the predominant functions altered by CDAtreatment were cell death and cellular assembly and organization. This included alterations in secretedproteins, endoplasmic reticulum and mitochondrial chaperones, antioxidant proteins, and enzymes involvedin fatty acid biosynthesis. Comparative in vitro dosing studies showed similar alterations to the proteome,neutral lipid accumulation, and mitochondrial dehydrogenase activity in response to CDA treatment ofcultured rat hepatocytes. The finding that several proteins showed significant changes in abundance beforethe onset of overt toxicity in vivo suggested that these could serve as predictive biomarkers of compoundswith a propensity to induce liver steatosis. These markers underwent further direct analysis in the invitro hepatocyte toxicity model to determine their utility in the development of high throughput assaysfor drug-induced steatosis.