Extraction And Characterazition Of Phospholipids Profile And The Evaluation Of Proliferation Effection Of HepG2 Cells By Phosphatdylcholine Derived From Large Yellow Croaker Roe

The large yellow croaker(Pseudosciaena crocea)is one of the important cultured marine fish in the China's South-East coastal areas,which belongs to the Sciaenidae.In particular,Fujian has the greatest number of cultured P.crocea and the highest production.However,the by-products of fish roe has not utilized fully in current.In this project,we firstly analyzed the phospholipds content,fatty acids composition and phospholipids components in the different part of P.crocea.Afterwards,the phospholipids molecular species and the purification conditions of phosphatidylcholine from fish roe were investigated.Eventually,the effect of phosphatidylcholine lipidosome on multiplication of HepG2 cells was performed,which was prepared from P.crocea roe phosphatidy-lcholine.The main research results were as follows.1.The phospholipids components and its fatty acids composition in the different part of P.crocea.were analyzed.Results showed that the highest phospholipids content is roe(5.50%),and the second is the fish innards(2.40%).The phospholipids components research results showed phospholipids components in head,back muscle and abdominal muscle were accordance,and they were lysophosphatidylcholine(LPC),phosphatidylcholine(PC).The phospholipids components in visceral were LPC,phosphatidylinositol(PI)and sphingomyelin(SM).The phospholipids components in fish tail were LPC,PC and phosphatidyl-ethanolamine(PE).The phospholipids components in roe were LPC,PI,PE and PC.The content of PC in roe were the highest(76.36%).The analysis of fatty acids composition showed that the P.crocea.was rich in n-3 polyunsaturated fatty acids.The polyunsaturated fatty acids in roe accounted for 43.1%,and the content of both DHA and EPA was 41.90%.So roe was better obviously than other parts of the P.crocea..2.The phospholipids molecules in fish roe were identified and analyzed.The results showed that the phospholipids group contained eight kinds of phospholipids molecules.A total of 96 phospholipids including 49 phosphatidylcholines(PCs),3 lysophosphatidylcholines(LPCs),13 phosphatidylethanolamines(PEs),10 phosphatidic acids(PAs),13 phosphatidylserines(PS),three phosphatidylglycerols(PGs),two sphingomyelins(SMs)and two phosphatidylinositols(PIs)were deduced.The highest content of phospholipids components is PC(65.16%).The main molecules of PC were PC 15:0/19:1&16:0/18:1(19.43%),PC P-16:0/20:5&16:0/20:5(11.52%),PC O-16:0/22:6&P-16:0/22:6&16:0/22:6(13.33%)and PC O-18:0/22:6&18:0/22:6(15.79%).The unsaturated fatty acids of roe phospholipids given priority to PUFAs,especially DHA(C22:6),EPA(C20:5).3.The phosphatidylcholine(Roe-PC)from phospholipids in P.crocea roe was separated and purified by the column chromatography.The parameters of kinds of stationary phase,granularity,elute rate,eluant composition and loading amount were performed as indexes for optimizing the purification process.The results were shown that the optimum purification conditions were as follow,silica gel 200?300,chloroform/methanol 3:2,flow rate 2 mL/min,loading amount 0.035 g/g(sample/silica gel).Under these conditions,the purity of PC from P.crocea roe can be reached 96.34%,and the recovery rate was up to 85.18%.4.The Roe-PC liposome was prepared,while the Roe-PC prepared with the optimal purification conditions.The multiplication effection of Roe-PC liposome on HepG2 cells was preliminary explored.The toxicological effection of the Roe-PC liposome on HepG2 cells under different concentrations and time was determined by MTT method.Results showed that the concentration range 0 ?g/mL and 50 ?g/mL will not cause toxicity to cells.While it could cause toxicity to the cells when the concentration more than 100 ?g/mL.So it showed that Roe-PC liposome had certain anticancer effect to HepG2 cells.It had establish the foundation of science for subsequent in-depth study on the molecular mechanism.In conclusion,the basic features on the phospholipids components and fatty acids composition in the different part of P.crocea.were studied systematically.The phospholipids in fish roe and phospholipids molecules were investigated mainly on this basis.And the purification process of roe phosphatidylcholine was established and the optimal purification condition of phosphatidylcholine was obtained.Eventually,the multiplication effection of Roe-PC liposome on HepG2 cells was preliminary explored.Research findings established scientific foundation for the further development of marine life resources on fish roe.