high-throughput screening of australian marine organism extracts for bioactive molecules affecting the cellular storage of neutral lipids高通量筛选的澳大利亚海洋生物提取物生物活性分子影响中性脂肪细胞储存的.pdfVIP

High-Throughput Screening of Australian Marine Organism Extracts for Bioactive Molecules Affecting the Cellular Storage of Neutral Lipids 1 2 2 1 2 1 James Rae , Frank Fontaine , Angela A. Salim , Harriet P. Lo , Robert J. Capon , Robert G. Parton , Sally Martin1* 1 Division of Molecular Cell Biology, Institute for Molecular Bioscience, The University of Queensland, Brisbane, Queensland, Australia, 2 Division of Chemical and Structural Biology, Institute for Molecular Bioscience, The University of Queensland, Brisbane, Queensland, Australia Abstract Mammalian cells store excess fatty acids as neutral lipids in specialised organelles called lipid droplets (LDs). Using a simple cell-based assay and open-source software we established a high throughput screen for LD formation in A431 cells in order to identify small bioactive molecules affecting lipid storage. Screening an n-butanol extract library from Australian marine organisms we identified 114 extracts that produced either an increase or a decrease in LD formation in fatty acid-treated A431 cells with varying degrees of cytotoxicity. We selected for further analysis a non-cytotoxic extract derived from the genus Spongia (Heterofibria). Solvent partitioning, HPLC fractionation and spectroscopic analysis (NMR, MS) identified a family of related molecules within this extract with unique structural features, a subset of which reduced LD formation. We selected one of these molecules, heterofibrin A1, for more detailed cellular analysis. Inhibition of LD biogenesis by heterofibrin A1 was observed in both A431 cells and AML12 hepatocytes. The activity of heterofibrin A1 was dose dependent with 20 mM inhibiting