Abstract
Plasmalogens play multiple roles in the structures of biological membranes, cell membrane lipid homeostasis and human diseases. We report the isolation and identification of choline plasmalogens (ChoPlas) from swine liver by high performance thin layer chromatography (HPTLC) and high performance liquid chromatography (HPLC)/MS. The growth and viability of hepatoma cells (CBRH7919, HepG2 and SMMC7721) was determined following ChoPlas treatment comparing with that of human normal immortal cell lines (HL7702). Result indicated that ChoPlas inhibited hepatoma cell proliferation with an optimal concentration and time of 25 μmol/L and 24 h. To better understand the mechanism of the ChoPlas-induced inhibition of hepatoma cell proliferation, Caveolin-1 and PI3K/Akt pathway signals, including total Akt, phospho-Akt(pAkt) and Bcl-2 expression in CBRH7919 cells, were determined by western blot. ChoPlas treatment increased Caveolin-1 expression and reduced the expression of phospho-Akt (pAkt) and Bcl-2, downstream targets of the PI3K/Akt pathway. Further cell cycle analysis showed that ChoPlas treatment induced G1 and G1/S phase transition cell cycle arrest. The expression of essential cell cycle regulatory proteins involved in the G1 and G1/S phase transitions, cyclin D, CDK4, cyclin E and CDK2, were also analyzed by western blot. ChoPlas reduced CDK4, cyclin E and CDK2 expression. Taken together, the results indicate that swine liver-derived natural ChoPlas inhibits hepatoma cell proliferation associated with Caveolin-1 and PI3K/Akt signals.
Highlights
Plasmalgens are a unique subset of phospholipids in which the sn-1carbon of the glycerol backbone contains a vinyl ether– linked long chain hydrocarbon instead of the typical ester-linked fatty acid
To further confirm the high performance liquid chromatography (HPLC) result, an optimized high performance thin layer chromatography (HPTLC) method was used for choline plasmalogens (ChoPlas) separation (Figure 1B)[21]
They constituted 15-20% of total phospholipids in cell membranes and mainly distributed in heart, brain, kidney, lung and skeletal muscles whiles there are the lowest amounts in liver, which could be explained by their synthesis in liver [3,37] .The biological function of plasmalogens and their implication in diseases has remained elusive, the function of exogenous natural liver –derived plasmalogens in human health
Summary
Plasmalgens are a unique subset of phospholipids in which the sn-1carbon of the glycerol backbone contains a vinyl ether– linked (a cis double bond adjacent to an ether bond) long chain hydrocarbon instead of the typical ester-linked fatty acid. The aliphatic moieties at the sn-1 position consist of C16:0 (palmitic acid), C18:0 (stearic acid) or 18:1 (oleic acid) carbon chains, whereas the sn-2 position is occupied by polyunsaturated fatty acids (PUFA) and the head group is usually either ethanolamine (ethanolamine plasmalogens, EtnPlas) or choline (choline plasmalogens, ChoPlas) [1]. These structural and compositional features provide novel properties to plasmalogens and they represent up to 20 % of the total phospholipid mass in humans, their physiological roles have been challenging to identify and are likely particular to different tissues, metabolic processes and developmental stages[2]. The lowest amounts of plasmalogen are found within the liver, possibly owing to their synthesis in the liver and subsequent transport by lipoproteins to other tissues [3]
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