| Oxidation is the one of major reasons affecting the aging process and decrease in memory lead. Phosphatidylserine(PS)can promote learning, enhance memory and improve the role of age-related diseases. Can PS at a certain concentration increase the ability of antioxidant in hippocampal neurons? We studied the antioxidant and anti-apoptotic role of PS on hippocampal neurons.1. Determine PS extracted from brains by thin-layer chromatographic (TLC) We extracted PS from brains (cattle, sheep, chicken and fish), and also determined the nature and the amounts by thin-layer chromatographic (TLC) using V(Chloroform):V(Ethanol):V(Triethylamine):V(H2O)=10:11.3:11.7:2.7 as agent, and 0.3% Bromothymol as reagent. The content of PS in every 100g were separately 20.5g, 19.3g, 18.7g and 17.0g. It was simple and had high extraction rate and high sample recovery in this method by thin-layer chromatographic.2. PS extracted,separated and purificated from brains of animals We used Organic Soluent Extraction getting rid of 95% cholesterin of brains by Acetone, and extracted 99% lecithoid by n-Hexane and Diethyl ether. Then the substance was mixed round according to V(Diethyl ether):V(NaCO3)=1:1~4, freezed and delamination still secondly . The flavicant solution was PS. It was onefold by thin-layer chromatographic.3. Analyse the kinds of fatty acid After analysing PS extracted from above four animals by Armor esterification and GLC, we could make conclusion that it contained more unsaturated fattery acid of PS by freezing than desiccation. Above four animals brains all contained saturated fattery acid (14:0, 18:0), single unsaturated fattery acid (16:1, 18:1), n-3 unsaturated fattery acid (18:3n, 20:5n, 22:6n) and n-6 unsaturated fattery acid (18:2n, 20:2n, 20:3n, 20:9n, 22:4n, 22:5n). Unsaturated fattery acid in brains of fish came to 52.24.4. PS prevents the brain related diseases caused by H2O2About research of learning, memory and brain diseases ,most people believed the labyrinthic experimentation made by Morris (1984). Animals was grouped eight. negative control (A), 15mg H2O2/Kg/time/ day (B),30mg H2O2/Kg/ time/ day (C) and 30mg PS/Kg/time/day (D). Another two groups were first given 15mg H2O2/Kg /time/day (E) and 30mg H2O2/Kg/time/day (F), after 6h, they were given same PS. Last two groups were first given 15mg PS/Kg/time/day (G) and 30mg PS/Kg/time/day (H), after 6h, they were given same H2O2. The results were as follow:①Contrast D and A, the mice of D achieved the end point with shorter time and distance (P<0.05). Contrast C,B and A, the mice of B and C with longer time and distance came to the end point (P<0.01). Contrast E, F, G, H and A, the time and distance were longer than A.②Observe centrum cells of hippocampal neurons by HE Acording to grade of dead cells (Satoru,set al), A,D was grade 1, B, E, F, H were grade 2, C was grade 3.③The change of components in brains⑴The contant of lipofuscin (u/g). A was 219.2±7.5, C was 422.6±87.2, B,E,F,G,H were 249.8±7.5~343.6±72.3. But D was 93.7±38.5. All showed PS at a certain concentration could increase the ability of antioxidant and the ability of oxidant of H2O2.⑵Activation of MAO-BA was 45, C was 100, and D was 35. B,E,F,G,H were 29~72. It proved that H2O2 increased monoamine oxidase, PS reduced the ability of monoamine oxidase.⑶Content of AchA was 14.3±3.22, C was 8.76±1.88, and D was 17.89±3.71. B,E,F,G,H were 11.56±2.13~14.3±3.22. It proved that H2O2 increased the content of Ach, PS reduced it.5. PS can effectively inhibit neuronal apoptosis caused by H2O2 Hippocampal neurons were separately inbuted with PS 1.5, 15, 150mg/L for 24h,then 0.01/L H2O2 was in for 24h.At last we determined cells apototic ratio.They were separately 27.17±2.05, 16.17±2.21, and 37.17±2.45. It proved that PS could effectively inhibit neuronal apoptosis.Test concentration of Ca2+ by Confocal, and the concentration were separately 4.03±1.24, 24.50±2.12, and -(15±3.02). All indicated that PS had effect on concentration of Ca2+, but too much PS was damaged to cells.Innovative:1) We provided a practical separation parameters of brain PS by TLC.2) H2O2 could make damage to brain by oxidation, and PS at a certain concentration could increase the ability of antioxidant in hippocampal neurons by the labyrinthic experimentation.3) We have studied the antioxidant and anti-apoptotic of PS on hippocampal neurons in the cells level, and achieved the desired results that the effection was correlation with concentration of Ca2+.
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