| Objective: In this study, through the establishment of hyperlipidemia rats to ileum Na~+ / bile acid transporter as the research object of ileal Na~+ / bile acid transporter and bile acid metabolism and hyperlipidemia in the formation of the correlation, for the prevention and control high fat hyperlipidemia disease providing a new way of thinking.Method: Take Wistar rats ( male, weight 145±8.5g) 60 rats were randomly divided into 2 groups: normal diet control group, high fat diet experimental group. Be compared to normal diet control group, the experimental group to high fat diet . (Formula: in the basal feed Concentration 0.04 cholesterol, 0.1 lard, 0.05 sucrose, 0.005 sodium cholate, 0.002 propylthiouracil), through 90 days of feeding, the establishment of hyperlipidemia model, using of automatic biochemical analyzer blood cholesterol testing regularly, monit oring the blood cholesterol levels four to six weeks, observe whether the requirements modeling. Two groups which selected required to exclude non-Winstar type rats, whether eating food with other drugs, hyperlipidemia, diabetes, deformities or other diseases . After the success of mode ling, experimental methods used were divided into the following two aspects:1. Extraction of the control group and experimental group ileal intestinal tissue and colon tissue of RNA, reverse transcription polymerase chain reaction detection of two model ileal intestinal tissue of IBAT gene expre- ssion intensity. 2. Extraction of the control group and experimental group ileal intestinal tissue, paraffin sections by indirect immunofluorescence after staining were detected ileal intestinal tissue model of the IBAT expression intensity. Statistical data used SPSS14.0 for windows statis- tical package X2,Fish paired t test and exact test for statistical analysis, P <0.05 indicate statistical significance. Result: 1. Electrophoresis results showed that: the experiment Ileum PCR products have emerged internal referenceβ-actin gene of 355bp amplified 220bp band and IBAT gene was amplified with the experimental group, with a brightness of IBAT gene amplification was significantly stronger than the control group. Semiquantitative results analysis: calculated using quantity One software more lanes in each PCR product band of optical density, calculated IBAT gene andβ-actin band of the optical density of the relative ratio of IBAT gene by comparing the relative value of the sample, eventually come to IBAT gene expression in ileal tissue mRNA relative content. Semiquan- titative results of the relative optical density of each of the subjects in each group required the same processing conditions are determined to repeat 3 times, whichever is the final average of data for statistical use. The final results: the experimental group gene mRNA expression in rat IBAT higher Jejunum end(1.12817±0.050841) ,Ileum end(1.14613±0.061868),caecum(1.14320±0.048016),control rat IBAT gene mRNA expression was lower Jejunum end (0.88427±0.041664),Ileum end(0.92013±0.049935),caecum(0.86777±0.0336157),the expression of significant differences was statistically significant (t=-25.6988, t=-15.446 t=-24.007 P <0.05).2 Indirect immunoflu- orescence staining showed that: in the high-powered microscope, fluore- scence microscope ileal tissue particles appear yellow green staining positive cells. 5 randomly selected for each slice view, each view to obse- rve the 100 cells,≥20% positive. Use tables from the Fisher exact prob- ability test results show: the experimental group IBAT expression was 66.3%,78.5%,55.3%; the control group IBAT expression was 10.3%,12.5%,8.95% . The difference between them was significant, there was significant (X2=9.631 ,X2=9.873,X2=9.209 P <0. 05).Conclusion: 1. Experimental group of IBAT mRNA gene expression more than control group increased. 2. Experimental group and control group in the rat colon no IBAT gene expressi on. 3.Ileal experim- ental group was significantly higher expression of IBAT, so ileum bile acid active trans port process of enhanced bile acid uptake was increased, theincreaseingin cholesterol and lipid absorption, accelerate the formation of hyperli pidemia... |
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