Journal
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
Volume 68, Issue 51, Pages 15123-15133Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.jafc.0c04649
Keywords
miR-145; hnf4 alpha; LC-PUFA biosynthesis; Siganus canaliculatus
Funding
- National Natural Science Foundation of China [31702357]
- National Key R&D Program of China [2018YFD0900400]
- Natural Science Foundation of Guangdong Province [2020A1515011252]
- STU Scientific Research Foundation for Talents [NTF19019]
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Fish, particularly marine species, are considered as the major source of long-chain polyunsaturated fatty acids (LC-PUFA) in the human diet. The extent to which fish can synthesize LC-PUFA varies with species and is regulated by dietary fatty acids and ambient salinity. Therefore, to enable fish to produce more LC-PUFA, comprehending the mechanisms underlying the regulation of LC-PUFA biosynthesis is necessary. Here, the regulatory roles of miR-145 were investigated in the marine teleost rabbitfish Siganus canaliculatus. The hepatic abundance of miR-145 was lower in rabbitfish reared in low salinity (10 ppt) in comparison with that of those cultured in seawater (32 ppt), while the opposite pattern was observed for the transcripts of the transcription factor hepatocyte nuclear factor 4 alpha (Hnf4 alpha), known to affect rabbitfish LC-PUFA biosynthesis. Rabbitfish hnf4 alpha was identified as a target of miR-145 by luciferase reporter assays, and overexpression of miR-145 in the S. canaliculatus hepatocyte line (SCHL) markedly reduced the expression of Hnf4 alpha and its target genes involved in LC-PUFA biosynthesis, namely, Delta 4 fads2, Delta 6 Delta 5 fads2, and elovl5. The opposite pattern was observed when miR-145 was knocked down in SCHL cells, with these effects being attenuated by subsequent hnf4 alpha knockdown. Moreover, increasing endogenous Hnf4 alpha by the knockdown of miR-145 increased the expression of LC-PUFA biosynthesis genes and enhanced the synthesis of LC-PUFA in both SCHL cells and rabbitfish in vivo. This is the first report to identify miR-145 as a key effector of LC-PUFA biosynthesis by targeting hnf4 alpha, providing a novel insight into the mechanisms of the regulation of LC-PUFA biosynthesis in vertebrates.
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