The Studies Of Fish's Liver Injury Protection On Phyllanthus Urinaria L.

In order to study the fish's liver injury protection on Phyllanthus urinaria L.. Oreochromis niloriczcs have been researched by the following five parts in this paper.Part 1 The determination the gallic acid content of the different extracts of Phyllanthus urinaria L.Phyllanthus urinaria L.were extracted on water,ethanol and acetone.The gallic acid content of the different extracts of Phyllanthus urinaria L.were determination by High Performance Liquid Chromatography(HPLC)assay.The results showed that gallic acid content of water extracts(9.473 mg.g^-1)is the highest.Part 2 The optimum conditions of extracting polysaccharides of protecting liver substance from Phyllanthus urinaria L.The extraction methods have water,ethanol and acetone extraction.The extraction methods were selected with the gallic acid content and aminotransferase as the standard.During the extracting process,four major experiment factors were selected in single factor analysis method with the gallic acid content as the standard, including the extraction time,the extraction temperature,the ratio of material to liquid and extraction times.The best method was Water extraction.With the above conditions,the optimal processing conditions for polysaccharides extraction are obtained by orthogonal experiments as follows:the extraction time is 45 min,the extraction temperature is 100℃and the ratio of material to liquid is 1/30.Part 3 Effects of Phyllanthus urinaria L.to liver injury and immunostimulating activity of Oreochromis niloriczcs.In order to study effects of Phyllanthus urinaria L.to liver injury and immunostimulating activity of Oreochromis niloriczcs,Oreochromis niloriczc were fed with formulated diet supplement with 0.5%,1.0%,1.5%the distilling substance of Phyllanthus urinaria L.running for 7 ds.Iiver injury indexes(ALT,AST,ADA,LDH) and immunity indexes(AKP,ACP,MPO,lysozyme)are analyzed on after of CuSO₄·5H₂O poisoning 24 h respectively.The results showed that liver injury indexes and immunity indexes of Oreochromis niloriczcs extremely significant compared with control group(p＜0.01).Diets supplementation with different the distilling substance of Phyllanthus urinaria L.no significant effects were observed(P＞0.05).In conclusion,feed supplementeds with 0.5～1.5%the distilling substance of Phyllanthus urinaria L.could bate liver injury and improve immunity function of Oreochromis niloriczcs.Part 4 Effects of Phyllanthus urinaria L.to antioxidant activity and cell membrane function of Oreochromis niloriczcsIn order to study effects of Phyllanthus urinaria L.to liver injury and immunostimulating activity of Oreochromis niloriczcs,Oreochromis niloriczcs were fed with formulated diet supplement with 0.5%,1.0%,1.5%the distilling substance of Phyllanthus urinaria L.running for 7 ds.Antioxidant indexes(CAT,SOD,T-AOC,MDA)and cell membrane indexes(GSH-PX,Na⁺K⁺-ATPase)are analyzed on after of CuSO₄·5H₂O poisoning 24 h respectively.At the same time,the liver histological changes are observed.The results showed that antioxidant indexes and cell membrane indexes of Oreochromis niloriczcs extremely significant compared with control group (p＜0.01).Diets supplementation with different the distilling substance of Phyllanthus urinaria L.no significant effects were observed(P＞0.05).The pathologically examination was shown that in fish treated with Phyllanthus urinaria L.,degenerative and necrotic changes of the liver cells were obviously milder than those of fish untreated,above them the effect of the high dose group is the best.In conclusion,feed supplementeds with 0.5～1.5%the distilling substance of Phyllanthus urinaria L. could bate liver injury and improve antioxidant ability and cell membrane function of Oreochromis niloriczcs.Part 5 Effects of Phyllanthus urinaria L.to liver cells injuryIn order to study the Phyllanthus urinaria L.on liver cells injury protection mechanism.Application of cell isolation and culture application,single cell gel electrophoresis application,the observation effects of Phyllanthus urinaria L.to the liver cells on the P450 content,NO content,DNA damage.The results showed that content of P450 and NO,the "tail" appearance rate and the total length of the comet of liver cells of Oreochromis niloriczcs extremely significant compared with control group(p＜0.01).Diets supplementation with different the distilling substance of Phyllanthus urinaria L.no significant effects were observed(P＞0.05).In conclusion, Phyllanthus urinaria L.has the function of protecting liver cells.