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Effect of Cholesterol-bile Acid Metabolism in New Zealand White Rabbit Fed by Cholesterol-rich Diet with Fufang Jiangzhi No 3

Author: MaoQiZuo
Tutor: SunXu；QiuDongNi；LiuZuo
School: Fudan University
Course: Internal Medicine
Keywords: Cholesterol Bile acid Hypercholesterolemia High cholesterol diet The Fufang Jiangzhi No. 3 FXR CYP7A1 SHP BSEP LDL
CLC: R285.5
Type: Master's thesis
Year: 2009
Downloads: 87
Quote: 1
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Abstract


The first part of the compound lipid-lowering No. 3 New Zealand white rabbits, high-cholesterol diet, total cholesterol, LDL cholesterol and bile acid levels Objective: To observe the the compound lipid-lowering 3 New Zealand white rabbits, high-cholesterol diet serum total cholesterol, low-density lipid protein cholesterol and bile acid levels. Methods: 24 healthy male New Zealand white rabbits were divided into three groups: control group, the hypercholesterolemia model group (model group) as well as compound lipid-lowering intervention group (intervention group). The control group of rabbits with the conventional rabbit food four weeks of feeding; model group, the intervention group of rabbits with high-cholesterol diet (provided by the Shanghai Medical College, Fudan University Laboratory Animal Center, containing 1% cholesterol) for 4 weeks to cause hypercholesterolemia. Modeling at the same time, New Zealand white rabbits given the intervention group every morning to give half a pack of No. 3 Compound lipid-lowering (by gavage (Health Astragalus, Alisma of berberine, turmeric, raw Puhuang, hawthorn and malt Qiwei medicine) / 20mL distilled water) to observe the intervention. The observed indicators include rabbit ear vein serum total cholesterol level (CHO), low-density lipoprotein cholesterol levels (LDL-C), and bile acid level (BA). Results: process modeling, none of the three groups of White Rabbit produce diarrhea or constipation phenomenon. Modeling the end of four weeks, the control group, the model group and the intervention group, serum total cholesterol levels were 4.51 ± 0.45 mmol / L, 23.14 ± 3.21 mmol / L and 16.21 ± 2.14mmol / L. Model group, serum total cholesterol levels of the intervention group compared with the control group rose (P lt; 0.01), serum total cholesterol levels of the intervention group compared with the model group decreased (P lt; 0.01). Control group, the model group and the intervention group, serum low-density lipoprotein cholesterol levels were 1.37 ± 0.27mmol / L, 18.26 ± 2.51mmol / L and 11.80 ± 1.98 mmol / L,. Model group, the intervention group serum low-density lipoprotein cholesterol levels rose compared with the control group (P lt; 0.01), serum low-density lipoprotein cholesterol levels of the intervention group compared with the model group decreased (P lt; 0.01). Bile acid levels of the control group, the model group and the intervention group were of 7.45 ± 1.33μmol / L, 13.95 ± 1.86μmol / L and 13.31 ± 1.27μmol / L model group bile acid levels rose compared with the control group (P lt; 0.01); bile acid levels of the intervention group compared with the model group increased (P lt; 0.01); model group and the intervention group bile acid levels showed no statistical significance (P gt; 0.05). Discussion: Compound lipid-lowering can reduce high cholesterol diet induced New Zealand white rabbits with hypercholesterolemia serum total cholesterol and low-density lipoprotein cholesterol levels. The second part of the compound lipid-lowering No. 3 New Zealand white rabbits on a high cholesterol diet the liver CYP7A1mRNA level and CYP7A1 activity purpose: To observe the impact of the Fufang Jiangzhi No. 3 New Zealand white rabbits, high-cholesterol diet, the liver CYP7A1 mRNA level and CYP7A1 activity, explore Compound lipid-lowering regulate cholesterol - bile acid metabolism mechanism of action to lower blood cholesterol levels. Methods: modeling and intervention methods with the first part. Four weeks after the end of the experiment, the use of air embolism anatomical experimental white rabbits were sacrificed and liver samples taken from the same site, immediately cryopreservation (first placed temporarily stored in liquid nitrogen, transfer to the refrigerator at -80 ℃ save). CYP7A1 mRNA expression in the liver tissue with real-time PCR assay, the activity of CYP7A1 in the liver tissue was measured by enzyme-linked immunosorbent assay. Results: process modeling, none of the three groups of White Rabbit produce diarrhea or constipation. Real-time quantitative PCR assay CYP71 mRNA expression in liver tissue, the experimental results: CYP7A1 mRNA expression in the liver tissue of the control group, the model group and the intervention group were 1.97 ± 1.50,0.70 ± 0.27 and 3.87 ± 1.68. The model group CYP7A1mRNA compared with the control group decreased (P lt; 0.01); the intervention group CYP7A1mRNA elevated compared with the control group (P lt; 0.01); the intervention group CYP7A1mRNA compared with the model group increased (P lt; 0.01). By enzyme-linked immunosorbent assay the activity of CYP7A1 in the liver tissue, the experimental results: the concentration of CYP7A1 in the liver tissue of the control group, the model group and the intervention group were 0.80 ± 0.22μg/ml, 0.44 ± 0.16μg/ml and 0.81 ± 0.21 μg / ml. CYP7A1 activity group model group compared with the control group decreased (P lt; 0.01); CYP7A1 activity of the intervention group and the control group showed no statistical significance (P gt; 0.05); increased CYP7A1 activity group intervention group compared with the model group (P lt; 0.01 ). Discussion: The results show that: the model group CYP7A1mRNA expression and CYP7A1 activity is lower than in the control group; the intervention group CYP7A1mRNA expression and CYP7A1 activity compared with the model group increased. Prompted a high-cholesterol diet caused elevated serum bile acid levels expression decreased CYP7A1mRNA of, CYP7A1 activity decreased inhibition classic bile acid synthesis pathway. No. 3 by the application of compound lipid-lowering promote CYP7A1mRNA the upregulation of CYP7A1 activity increased intrahepatic cholesterol to bile acid conversion increased, leading to the decline in the level of blood cholesterol. The third part of the compound lipid-lowering No. 3 New Zealand white rabbits, high-cholesterol diet, the liver of BSEP mRNA expression of SHP mRNA and LDL mRNA Objective: To observe the the Fufang Jiangzhi New Zealand white rabbits, high-cholesterol diet liver tissue BSEP mRNA SHPmRNA and LDL mRNA expression, and to further explore the mechanism of action of the 3rd compound lipid-lowering intervention effect. Methods: modeling and intervention methods with the first part. The liver specimen collection and save it with the second part. Using real-time fluorescence quantitative PCR assay test White Rabbit liver tissue BSEP mRNA in, SHP mRNA and LDL mRNA expression. Results: control group, model group, the intervention group of New Zealand White rabbits liver tissue BSEP mRNA expression is as follows: 7.33 ± 2.44,16.40 ± 3.01 and 9.64 ± 1.63. BSEP mRNA expression of model group compared with the control group BSEP mRNA expression was significantly higher (P lt; 0.01); BSEP mRNA expression intervention group compared with model group BSEP mRNA expression was significantly decreased (P lt; 0.01); higher than those in the control group BSEP mRNA expression (P lt; 0.01). Control group, model group, the intervention group of New Zealand White rabbits liver tissue SHP mRNA expression is as follows: 9.83 ± 2.5085.44 ± 6.79 and 20.73 ± 4.47. SHP mRNA expression in the model group compared with the control group SHP mRNA expression was significantly increased (P lt; 0.01); SHP mRNA expression in the intervention group compared with the model group SHP mRNA expression was significantly decreased (P lt; 0.01); higher than those in the control group SHP mRNA expression (P lt; 0.01). Control group, model group, the intervention group of New Zealand White rabbits liver tissue LDL mRNA expression is as follows: 5.76 ± 1.963.29 ± 1.58 and 9.38 ± 2.86. The model group LDL mRNA expression LDL mRNA expression was significantly decreased compared with the control group (P lt; 0.01); LDL mRNA expression in the intervention group compared with the model group, LDL mRNA expression was significantly higher (P lt; 0.01); also higher LDL mRNA expression (P lt; 0.01). Discussion: After four weeks of experimental the White Rabbit liver tissue FXR target genes BSEP mRNA and SHP mRNA expression by RT-PCR found SHP mRNA model group and BSEP mRNA compared with that in the control group was significantly higher (P lt; 0.01), prompted FXR activity increased in liver tissue of the model group. Use the 3rd compound lipid-lowering intervention, the intervention group SHP mRNA and BSEP mRNA compared with the model group decreased (P lt; 0.01), prompted the intervention group liver tissue FXR activity compared with the model group decreased, indicating that the compound lipid-lowering in the 3rd weakened FXR activation to regulate cholesterol - bile acid metabolism, which is play a main mechanism of the intervention effect of hypercholesterolemia.

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