Effects of ELOVL4 gene overexpress on the synthesis efficiency of n3 and n6 very long chain polyunsaturated fatty acids / 国际眼科杂志(Guoji Yanke Zazhi)

ABSTRACT

AIM:To compare the synthesis efficiency of n3 and n6 very long chain polyunsaturated fatty acid ( VLC-PUFA ) by overexpressing ELOVL4 protein, providing guidance for treating Stargardt-like macular dystrophy (STGD3). METHODS:To establish recombinant adenovirus with the ELOVL4 protein and green fluorescent protein, transferred into cultured PC12 cells. The cells were divided into 3 groups PC12, PC12 + Ad- GFP and PC12 + Ad-ELOVL4, former two groups serve as controls. ELOVL4 gene expression was quantified by qRT-PCRs. ELOVL4 protein was analyzed by Western - Blot ( WB ) . The transduced cells were treated with both EPA and AA (11). After 48h of incubation, cells were collected, total lipids extracted and fatty acid methyl esters prepared and analyzed by gas chromatography-mass spectrometry ( GC-MS) . RESULTS:When supplemented together, 205n3 (EPA) and 204n6 ( AA) were efficiently taken up at almost the same amounts in the PC12 cells regardless of ELOVL4 expression. The ELOVL4-expressing cells elongated both EPA and AA to a series of n3 and n6 VLC-PUFAs. From 205n3/EPA, 345n3 and 365n3 account for 0. 71% and 1.6%, respectively. From 204n6/DHA, 344n6 and 364n6 were only 0. 46% and 0. 61%, respectively. The total relative mol% of n3 VLC-PUFAs synthesized from EPA was almost two times that of n6 VLC-PUFAs synthesized from AA. CONCLUSION: ELOVL4 protein preferentially elongates n3 PUFA to VLC - PUFAs over n6 PUFA. Dietary supplementation of appropriate n3/n6 PUFAs may provide STGD3 patients with some therapeutic benefits.