Bsep And Ntcp Under The Control Of FXR In Lipid Metabolism In Rats With Bile Acids And The Role Of Cholesterol

Abstract:Objective: The rat model of hyperlipidemia through to farnesyl derivatives X receptor (FXR) and the FXR target genes - primary and secondary liver bile duct side of the liver cell membrane of capillary distribution of bile salt export pump (bile salt export pump, BSEP / Bsep), sinusoidal liver cells on the surface of the basement membrane sodium taurocholate transporter (sodium / taurocholate cotransporting polypeptide, NTCP / Ntcp) as the research object under the control of the FXR Bsep and Ntcp in hyperlipidemic disease and cholesterol metabolism in rats with bile acids in the formation and hyperlipidemia and the role of the correlation, for the prevention and treatment of hyperlipidemia to provide a new basis and molecular basis, is expected for the treatment of bile acid metabolism and related diseases to provide a kinds of new ideas. Methods: Wistar rats (male, weight 145±8.5g) 60 rats were randomly divided into 2 groups, n = 30: normal diet control group (control group), experimental high-fat diet group (experimental group). Control group were given normal diet, high fat diet to the experimental group (the formula is: in the basal diet added 0.04 cholesterol, lard 0.1, 0.05 sucrose, 0.005 sodium cholate, 0.002 propylthiouracil), fed 90 days established hyperlipidemia model, regular use of automatic biochemical analyzer blood cholesterol test, 8 to 12 weeks to monitor blood cholesterol levels, observe whether the requirements modeling. Rats are required to exclude selected groups with other drugs have been drinking diet, hyperlipidemia, diabetes, deformities and other diseases exist. After the success of modeling, the use of experimental methods into the following two areas: 1. To take control and experimental groups of liver tissue RNA, reverse transcription - polymerase chain reaction (RT-polymerase chain reaction; RT-PCR) to detect two group model of liver tissue FXR, Bsep and Ntcp gene expression intensity; 2. to take control and experimental groups of liver tissue, paraffin sections by immunohistochemistry after the SP (streptavidin-perosidase) model of liver tissue was detected by two groups of FXR, Bsep and Ntcp expression of strength. Statistics SPSS14.0 for windows statistical package used for statistical analysis, P <0.05 was considered statistically significant. Results: 1. Electrophoresis results showed that: the experiment liver PCR products have emerged in the internal reference geneβ-actin 425bp amplified band, and FXR, Bsep and Ntcp gene of 283bp, 289bp, 325bp amplified band, the experimental group and the FXR gene the regulation of gene amplification under the Bsep with brightness was stronger than the control group; Ntcp gene amplification in the experimental group was weaker than the control group with brightness. Semi-quantitative results of analysis: Comparison of Quantity One software to calculate the PCR products of each lane optical density of bands, calculated FXR, Bsep and Ntcp gene andβ-actin optical density of bands relative ratio of the sample by comparing the FXR, Bsep and Ntcp gene relative value, and ultimately come to FXR, Bsep and Ntcp mRNA gene expression in liver tissue relative content. Semi-quantitative results of the relative optical density value of each of the subjects in each group required the same treatment conditions were 3 times repeated measurements, whichever is the ultimate use of the data as a statistical average. Final results show: experimental group FXR, Bsep mRNA gene expression higher (0.8519±0.0270, t=65.398 P <0.05), (0.8641±0.0221,t=61.110 P<0.05), control rat Ntcp gene mRNA was higher than the experimental group (0.8645±0.0204,t=61.945 P<0.05). Differences between the expression of three genes, were statistically significant. 2. Immunohistochemical SP (streptavidin-perosidase) assay showed that: in the liver of high power microscope microscopic particles in a yellow staining positive cells. 5 randomly selected each slice view, each field of vision to observe the 100 cells,≥10% positive cells were positive, or negative, the use of tables from the Fisher exact probabilities test. The results showed that: the experimental group FXR, Bsep, Ntcp expression was 75%, 76.7%, 24.5%; control group FXR, Bsep, Ntcp expression was 17.3%, 18.5%, 71%. The difference between them was significant, there was significant FXR (X2 = 10.932 P<0.05), Bsep (X2 = 10.773 P<0.05), Ntcp (X2 = 9.759 P<0.05).Conclusion: 1. Experimental rats FXR gene mRNA expression was significantly increased compared with the control group. 2. Experimental group FXR gene under the control of Bsep mRNA expression in control group, Ntcp mRNA gene expression than the control group. Therefore, hyperlipidemia, liver bile acid excretion rate increased uptake reduction, slow down the formation of hyperlipidemia to reduce the burden on the liver and maintain liver function. 3. As FXR target genes, Bsep consistent with changes in FXR, Ntcp and FXR opposite directions. Prompt us to be committed to the development of drugs acting on FXR, hyperlipidemia and related diseases to find new treatment methods and means.