Proteomic and lipidomic analysis of primary mouse hepatocytes exposed to metal and 1 metal oxide nanoparticles
نویسندگان
چکیده
20 The global analysis of the cellular lipid and protein content upon exposure to metal and metal oxide 21 nanoparticles (NPs) can provide an overview of the possible impact of exposure. Proteomic analysis has 22 been applied to understand the nanoimpact however the relevance of the alteration on the lipidic profile 23 JOURNAL OF INTEGRATED OMICS A METHODOLOGICAL JOURNAL HTTP://WWW.JIOMICS.COM Journal of Integrated Omics has been underestimated. In our study, primary mouse hepatocytes were treated with ultra-small (US) 24 TiO2-USNPs as well as ZnO-NPs, CuO-NPs and Ag-NPs. The protein extracts were analysed by 2D-DIGE 25 and quantified by imaging software and the selected differentially expressed proteins were identified by 26 nLC-ESI-MS/MS. In parallel, lipidomic analysis of the samples was performed using thin layer 27 chromatography (TLC) and analyzed by imaging software. Our findings show an overall ranking of the 28 nanoimpact at the cellular and molecular level: TiO2-USNPs<ZnO-NPs<Ag-NPs<CuO-NPs. CuO-NPs and 29 Ag-NPs were cytotoxic while ZnO-NPs and CuO-NPs had oxidative capacity. TiO2-USNPs did not have 30 oxidative capacity and were not cytotoxic. The most common cellular impact of the exposure was the 31 down-regulation of proteins. The proteins identified were involved in urea cycle, lipid metabolism, 32 electron transport chain, metabolism signaling, cellular structure and we could also identify nuclear 33 proteins. CuO-NPs exposure decreased phosphatidylethanolamine and phosphatidylinositol and caused 34 down-regulation of electron transferring protein subunit beta. Ag-NPs exposure caused increased of total 35 lipids and triacylglycerol and decrease of sphingomyelin. TiO2-USNPs also caused decrease of 36 sphingomyelin as well as up-regulation of ATP synthase and electron transferring protein alfa. ZnO-NPs 37 affected the proteome in a concentration-independent manner with down-regulation of RNA helicase. 38 ZnO-NPs exposure did not affect the cellular lipids. To our knowledge this work represents the first 39 integrated proteomic and lipidomic approach to study the effect of NPs exposure to primary mouse 40 hepatocytes in vitro. 41 42 43
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