The Pharmacodynamic Function Study Of Sedum Sarmentosum Extract In Grass Carp Fatty Liver Model

1High-fat diet built grass carp fatty liver model We tested the healthy grass carp of12±1.5g weight into the basic group and high-fat group, experimented on feeding the normal physiological metabolism of4.6%(fish oil2.8%+1.8%soybean oil)basic diet and8.1%(5%fish oil+3.1%soybean oil) high fat diet of grass carp for six consecutive weeks. And on the weeks of second, fourth and6th, six grass carps of the basic group and high-fat group would be randomly selected to collecte serum and hepatopancreas tissue samples. We would observe the changes of the internal organs and acquire hepatopancreas tissue samples to explain reflect major the grass carp hepatopancreas injury extent physiological and biochemical indices and HE and Sudan staining observed judgment grass carp hepatopancreas degree of injury caused by the high-fat diet, to determine fatty liver model of the two groups of grass carp hepatopancreas. The results showed that:the test during the basic set of grass carp serum, physiological and biochemical indicators of liver and pancreas tissue and its histological pathological did not changed significantly. The high-fat group grass carp serum results of physiological and biochemical showed that:alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride (TG) and total cholesterol (CHO) levels along with the extension of the trial time was significantly increased (P<0.01), that explained to the excessive amounts of high-fat diet the liver cells damage of grass carp; resulting in increased serum TG and CHO content, explained to grass carp hepatopancreas accumulated excess lipids; the hepatopancreas organization malondialdehyde (MDA), superoxide dismutase (SOD) and catalase (CAT) level with the prolongation of the test time, were also showing a trend of significant increased (P<0.01). Description excess fat diet causes the body over oxidation reaction exacerbated by a serious of hepatopancreas injury. The histopathological observations displayed:The grass carp hepatopancreas of high fat group has found serious liver cell lipid droplets accumulated in the test on fourth weeks, but the liver cells of most serioutly steatosis occurred on the end of the sixth weeks of the test. Integrated the serum, hepatopancreas physiological and biochemical changes, and histopathological observations of the high-fat group grass carp could be considered feeding the grass carp of six consecutive weeks8.1%higt fat diets could be copied fatty liver model successfully.2The pharmacological effects of Sedum sarmentosum extracts to grass carp fatty liver The grass carp which were established fatty liver model successfully were divided into five groups, respectively, for the high-fat group(fed8.1%high-fat diet); the high-dose group of Sedum sarmentosum (fed8.1%high-fat diet+1.2g·kg-1Sedum sarmentosum extracts); Medium-dose group of Sedum sarmentosum (fed8.1%high-fat diet+0.6g·kg-1Sedum sarmentosum extracts); low-dose group of Sedum sarmentosum (fed8.1%high-fat diet+0.3g·kg-1Sedum sarmentosum extracts); treatment control group (fed8.1%high-fat diet+0.02g·kg-1Zhibituo); Used another healthy grass carp as a basis for group fed4.6%basic diet. Daily at9:00am,12:00am and6:00pm, fed the diet for six consecutive weeks. And at the weeks of second, fourth and6th, we selected six grass carp randomly at each of the group to collected serum and observed the changing of internal organs then collected hepatopancreas tissues. Observed the changes of the indexes that could reflect the degree of grass carp hepatopancreas damaged and using HE and Sudan to detect the function of treatment within6weeks. The results showed that: comparing with the high-fat group, the high dose group and the low dose group of Sedum sarmentosum could significantly reduce the levels of ALT, AST, TG, CHO, MDA and CAT(P<0.05) and increase the level of SOD(P<0.05) of the grass carp with fatty liver. The extract of Sedum sarmentosum can alleviated the grass carp liver and pancreas damage effectively, reduce blood lipid levels, enhance the antioxidant capacity of the hepatopancreas. The histopathological observed:Sedum sarmentosum extract could improve grass carp liver cell steatosis that were caused the continued high-fat diet with six weeks, and Sedum sarmentosum extract could promote lipid metabolism in the liver cells. Therefore the test of serum and hepatopancreas physiological and biochemical with all these groups of grass carp indicated the high dose group of the Sedum sarmentosum had the best comprehensive treatment works that could significant treatment the grass carp fatty liver of continue feeding high fat diet induced.3Grass carp SREBP-1、PPAR-α、TNF-a and CPT-1mRNA expression test The samples were from control goup, model group, treatment control gourp and medium-dose group respectively at6th week. The total RNA was extracted by Trizol. Using specific primers, the target gene were gotten by two-step RT-PCR. Semi-quantitative deteced the relative expression of SREBP-1, CPT-1, PPAR-a and TNF-a mRNA relative expression level. The results of semi-quantitative RT-PCR analysis showed that:comparing with the high-fat group, the grass carp of high dose group and the low dose group of Sedum sarmentosum could significantly elevate the levels of hepatopancreas CPT-1and PPAR-a mRNA relative expression(P<0.01), and reduce the level of high dose group sarmentosum SREBP-1and TNF-a mRNA expression, especially the high-fat group(P<0.01).