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The Establishment of Fatty Liver Model in Vitro and Vivo and Drug Screening

Author: PanXueFeng
Tutor: XuJianHua
School: Fujian Medical
Course: Pharmacology
Keywords: Curcumin derivatives Tetrahydrocurcumin Fatty liver In vitro In vivo
CLC: R96
Type: Master's thesis
Year: 2009
Downloads: 114
Quote: 1
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Abstract


Background and Purpose fatty liver manifestations of the metabolic syndrome, the early simple steatosis steatohepatitis, liver fibrosis and cirrhosis, and subsequently may also occur. Its high incidence of more and more attention. Actively studying the development of fatty liver treatment is efficient, inexpensive, convenient, safe drugs is significant. The topics to explore the establishment of fatty liver in vitro and animal models, the observed curcumin derivatives aeries JZ2 and, SFR1 tetrahydrocurcumin protective effects in vitro and in vivo models of fatty liver. Method (1) the the oleic role in HepG2 cells and normal liver cell line L02 established in vitro model of fatty liver cells,, MTT assay screening of curcumin derivatives aeries JZ2 and, SFR1 tetrahydrocurcumin role in liver cells non-cytotoxic thick degree, administered by different groups of non-cytotoxic concentration of intracellular lipid droplets formation and detection cell supernatant ALT, AST and gamma-GT Oil Red O staining were observed. To observe the effects of drugs in vitro anti-fatty liver. (2) by giving high fat diet, to establish a mouse model of fatty liver by intraperitoneal injection of tetracycline composite, given different doses of curcumin derivatives aeries JZ2 and at the same time, the SFR1 and tetrahydro curcumin solid dispersion experiment 36 days blood and liver tissue, liver index, serum lipids and liver function: serum TG, CHO, LDL-C, HDL-C, ALT, AST, liver lipid: the liver homogenate TG, CHO; frozen sections of the liver and Oil Red O staining biopsy means to observe the pathological changes of the liver. Thus observed drug the lipotropic efficacy in mice. 1 (1) by MTT screened curcumin derivatives aeries JZ2 SFR1 non cells and a concentration of 100 ug / ml and 200 ug / ml and 300 ug / ml, 400 ug / ml, tetrahydrocurcumin non Cells concentration of 10 ug / ml, 20 ug / ml, 30 ug / ml; (2) Oil Red O staining was found, in the light microscope after 24 hours of the 80 ug / ml oleic acid stimulates HepG2 cells, the cells of the model group There are a large number of red lipid droplets accumulate red lipid droplets started to decrease after 48 hours; 60 ug / ml oleic L02 cells 24 hours after stimulation, the cells of the model group, a lot of red lipid droplets accumulate red lipid droplets of cells after 72 hours started to decrease. Reduced interval of 24 hours between doses of drug action in HepG2 cells at 48 and 72 hours showed no lipid droplets; reducing drugs that act on the L02 cells 48 hours no lipid droplets; drugs that act on the L02 cells 72 hours with the role of increased concentration intracellular lipid droplets gradually reduced. Oil Red O staining Image analysis of drugs of oleic acid to stimulate 48h after L02 cells (3) with the ImagePro Plus6.0 image analysis software, each dose group compared with model group, the lipid droplets significantly reduced (P <0.05). Which tetrahydrocurcumin 30ug/ml JZ2300ug/ml, SFR1400ug/ml difference highly significant difference (P <0.01) (4) to detect cell supernatant in ALT, AST, γ-GT, that tetrahydrocurcumin on effect on liver function smaller, JZ2, SFR1, liver function have a certain influence. 2 (1) by giving a high fat diet, intraperitoneal injection of 150mg/kg tetracycline composite successfully established a mouse model of fatty liver, compared with the normal group, model group mice liver index, serum TG, CHO, LDL -C, ALT, AST, and liver homogenates TG, CHO were significantly higher (P lt; 0.05). Liver frozen sections of oil red O staining, liver biopsy and HE staining also confirmed its occurrence fatty liver disease; (2) compared with the model group, tetrahydrocurcumin solid dispersant on the CHO, LDL-C, HDL-C decreased, but more significant role in the low-dose group, a significant part of the data, the TG role is not obvious. The index in the liver and intrahepatic lipid especially TG, liver function ALT, AST have shown a drop in the role, and there is a significant dose-dependent relationship. Group 200mg/kg, 300mg/kg liver index, intrahepatic lipid (TG), improved liver function ALT statistically significant. Frozen sections with oil red 0 staining and pathological to prove tetrahydrocurcumin protective effect of fatty liver in mice. (3) Compared with model group, curcumin derivatives JZ2 high and low dose group on blood lipids TG, CHO, LDL-C reduction, significant part of the data, TG, HDL-C in the low dose group role better. SFR1 high and low doses of lipids CHO, different degrees of reduction of LDL-C, part of the data was statistically significant, but TG undesirable impact. Two drugs on mice liver index, liver lipid had no effect. The frozen sections Oil Red O staining and biopsy with the model group no significant difference. Conclusion (1) to establish a more stable fat liver in vitro sieve drug model: oleic stimulation L02 cells 48 hours, drug administration for the 24 hours before oleic stimulation once after 24 hours together with the oleic acid be administered once, and can be filtered drug in vitro anti-fatty liver. (2) the establishment of a mouse model of fatty liver: high fat diet, intraperitoneal injection of 150mg/kg tetracycline. (3) curcumin derivatives aeries JZ2 and, SFR1 tetrahydrocurcumin must intervene in the role of fatty liver in vitro model, effectively reducing the formation of intracellular lipid droplets smaller tetrahydro curcumin tetrahydrocurcumin the liver function affect JZ2, SFR1 on liver function. (4) curcumin derivatives aeries JZ2 and SFR1 in 1000mg/kg lipids to improve the role, but there is no protective effect of fatty liver. (5) tetrahydrocurcumin solid dispersant of lipids to improve the role, but more significant in the low-dose 100mg/kg role, has a protective effect on the fatty liver.

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