| Objective: In this study,the mice model induced by cholesterol induced stone diet was used to intervene with gentian Xiegan decoction.The changes of cholesterol and bile acid concentrations and the expressions of NPC1L1,ABCG5 and ABCG8 which are closely related to cholesterol absorption were compared in different groups of mice.The molecular biological basis of CS dampness-heat syndrome was studied from the Angle of cholesterol metabolism.Methods: Thirty-two male mice of SPF grade C57BL/6J were randomly divided into the blank group(8)and the model group(24).CS mouse model was reproduced by high cholesterol-induced stone feed.After 8 weeks of modeling,8 model mice were randomly selected for sampling to observe whether the gallstone model was successful.The mice were randomly divided into model group and Longdan Xiegan decoction group.The longdan xiegan decoction group was treated with longan Xiegan decoction,and the model group and the blank group were treated with normal saline of the same volume for 4 weeks.The gallbladder morphology of mice in the blank group was observed as the control group,and the liver and small intestine tissue morphology of mice in each group was observed by HE staining method.Serum TC,TG,bile TC and TBA were detected by ELISA.The mRNA and protein levels of MICE NPC1L1 and ABCG5/G8 were measured by Q-PCR and Western blot,respectively.Results:1.General situation comparison: during the modeling period,the growth rate of body weight in the model group was faster than that in the blank group(P < 0.01),while the amount of diet and drinking water in the model group was significantly lower than that in the blank group(P < 0.01).After the intervention of Longdan Xie Decoction,the growth rate of body weight in the model group was slower,and the weight in the model group was significantly lower than that in the blank group(P < 0.01).2.Results under the microscopeUnder the stereomicroscope: the gallbladder of the blank group was clear,the gallbladder of the model group was larger than that of the blank group,the bile was turbid,there was flocculent precipitate,the gallbladder of the Longdan Xie Decoction group was turbid,and the gallbladder of the model group was clear.Microscopically,the hepatocytes in the liver tissue of the blank group were arranged in the shape of cords,with normal structure,intact intestinal mucosa structure,clear outline,orderly and regular brush border arrangement,and thin muscular layer;the fatty granules in the liver tissue of the model group were obvious,the hepatocytes near the central vein were arranged in disorder,the villi structure of the small intestine was damaged,the villi were arranged in disorder,and the muscular layer was thickened;in the Longdan Xie Decoction group,the fatty vesicles in liver tissue decreased,the intestinal villi disorder in small intestine improved,and the thickness of muscle layer decreased.3.Blood lipid and bile content: compared with the blank group,the level of TC and bile TC,TBA in the model group was significantly increased(P < 0.01),while the level of TG in the model group was not significantly increased(P > 0.05).Compared with the model group,the levels of TC,TC,and TBA in serum in the Longdan Xie Decoction group decreasedsignificantly(P < 0.01),while TG in serum decreased significantly(P < 0.05).4.mRNA expression of cholesterol metabolism-related genes: compared with the blank group,the expression of ABCG5 / G8 in liver tissue and NPC1L1 in small intestine increased significantly(P < 0.01),while the expression of ABCG5 / G8 in liver tissue and small intestine decreased significantly(P < 0.01).Compared with the model group,the expression of ABCG5 in the liver and small intestine decreased significantly(P < 0.01),ABCG8 in liver decreased significantly(P < 0.05),while the expression of ABCG5 / G8 in the liver and small intestine increased significantly(P < 0.01).5.Cholesterol metabolism-related gene protein expression: compared with the blank group,ABCG5 expression in liver tissues was significantly increased(P<0.01),ABCG8 and SMALL intestine tissues were significantly increased(P<0.05),while NPC1L1 and small intestine tissues were significantly decreased(P<0.01).Compared with the model group,ABCG5 decreased significantly in the longdan xiegan decoction group(P<0.01),and the expression of NPC1L1 was down-regulated but not significantly decreased in the liver tissues and small intestine tissues(P > 0.05),while the expression of NPC1L1 and small intestine tissues was up-regulated but not significantly increased(P > 0.05).Conclusion:1.The fatty change of liver tissue and the damage of small intestinal mucosal structure in cholecystolithiasis may be the pathomorphologic changes of dampness and heat syndrome.2.Dyslipidemia and imbalance of bile salts may be one of the molecular biological bases of dampness and heat syndrome.3.Abnormal cholesterol metabolism caused by changes in the expression of cholesterol metabolis-related genes ABCG5,ABCG8 and NPC1L1 in gallbladder cholesterol stones is one of the molecular biological bases of dampness and heat syndrome. |
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