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Role of Wnt5b in fatty acid synthesis in primary cultured mouse hepatocytes (1037.6)
Author(s) -
Siriwardhalin,
Kim Jung Han
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.1037.6
Subject(s) - transfection , lipogenesis , paracrine signalling , autocrine signalling , lipid metabolism , fatty acid synthesis , biology , fatty acid , cell culture , biochemistry , wnt signaling pathway , metabolism , medicine , endocrinology , microbiology and biotechnology , gene , genetics , receptor
Wnt proteins are secreted glycoproteins that act in autocrine and paracrine fashions and known to regulate fundamental cellular functions including cell differentiation, cell growth and apoptosis. Emerging evidence indicates that Wnts are also involved in the development of metabolic disorders. Large‐scale gene expression analysis of liver of an obesity mouse model fed high fat diets displayed decreased expression of number of genes including Wnt5b. In this study, we investigated the role of Wnt5b in lipid metabolism using RNA interference in primary cultured mouse hepatocytes. Hepatocytes were isolated from C57BL/6J mice and transiently transfected with siRNA targeting mouse Wnt5b or control siRNA. Cells were then maintained in culture medium for 48 hours before assays. For fatty acid synthesis, cells were incubated with 3 H 2 O (0.5 mCi/ml) for 2 hours, followed by saponification. Fatty acids were extracted and the 3 H content of fatty acids determined. For fatty acid oxidation, cells were incubated with 22 M [9,10(n) 3 H] palmitate substrate for 1 hour and the degree of incorporation of 3 H into H 2 O determined. In the cells transfected with Wnt5b siRNA, fatty acid synthesis increased 1.9 fold compared with that in the control siRNA transfected cells (p<0.05). There were no differences in beta‐oxidation. These results may suggest that Wnt5b plays a role in hepatic lipogenesis.