Prevention And Treatment Effect And Its Mechanism Of Yinchenhao Decoction On Cholesterol Gallstones In Mice

Objective: To observe the effect of Yinchenhao decoction(YCHD)on cholesterol gallstones induced by lithogenic diet in mice and explore its possible mechanism.To provide a more scientific experimental basis for the clinical application of YCHD.Methods: Experiment one: Study on the prevention effect of YCHD on cholesterol gallstones in mice.A mouse model of cholesterol gallstone was established by feeding lithogenic diet.Male C57BL/6 mice were randomly divided into normal control group(N),model group(M),Yinchenhao decoction low dose group(YCHD-L),Yinchenhao decoction high dose group(YCHD-H),and positive control group(TUDCA).Corresponding treatments via intragastric administration were performed simultaneously with the model diet for consecutively four weeks and eight weeks.Experiment two: Study on the therapeutic effect of YCHD on cholesterol gallstones in mice.A mouse model of cholesterol gallstone was established by feeding lithogenic diet.Male C57BL/6 mice were randomly divided into N,M,YCHD-L,YCHD-H and TUDCA groups.Corresponding treatments via intragastric administration were performed after four-weeks model diet,for consecutively another four weeks and eight weeks.Observation items:(1)The presence of gallstones was examined visually and the grade of gallstones in mice was judged through double-blind method.(2)Serum liver function and lipids were observed,and hepatic lipids were detected by enzyme colorimetry.(3)Hematoxylin-eosin staining of liver tissue.(4)Biliary lipid compositions were measured by enzyme colorimetry in order to calculate cholesterol saturation index(CSI).(5)The expression levels of relative genes involved in cholesterol metabolism pathway in liver and small intestine were examined by RT-q PCR and WB.Results: Experiment one: Compared with the normal group,the body weight of the model group was significantly reduced while the liver weight and the ratio of liver weight and body weight were significantly increased(P<0.01).The existence of crystals or stones were observed in the gallbladder.The serum levels of ALT,ALP,TC and LDL-C were significantly increased(P<0.01).Compared with the control group,the cytoplasm of the model mice was loose,accompanied with inflammatory cell infiltration.The degree of cytoplasmic loosening of hepatocytes in each group was reduced.The increased TC level and decreased TBA level in bile result in a significant increase in CSI(P<0.01).The expression levels of cholesterol metabolism-related genes(ABCG5,ABCG8,SRBI,CYP7A1,NPC1L1,etc.)in the liver and small intestine of the model mice were abnormal,and TC levels in liver tissue was significantly elevated(P<0.01).Compared with the model group,YCHD treatment could reduce the liver weight and the ratio of liver weight and body weight(P<0.05).After four-weeks administration,there was a decreased trend of serum ALT,ALP,TC and LDL-C which was not statistically significant(P>0.05).The serum levels of ALT,ALP,TC and LDL-C were reduced after eight-weeks administration(P<0.05).The reduced level of TC and increased level of TBA in bile via YCHD treatment for eight weeks could lead to a lower CSI(P<0.01)and the condition of gallstones in mice was alleviated.The overexpression of ABCG5,ABCG8,SRBI and NPC1L1 in liver and small intestine could be inhibited after four-weeks and eight-weeks administration while the level of TC in liver tissue was also reduced,of which the high dose group had the best effect(P<0.01).Experiment two: Compared with the normal group,the body weight of the model group was significantly reduced while the liver weight and the ratio of liver weight and body weight were significantly increased(P<0.01).The existence of crystals or stones were observed in the gallbladder.The serum levels of ALT,AST,TBA,ALP,TC and LDL-C were significantly increased(P<0.01).Compared with the control group,the cytoplasm of the model mice was loose,accompanied with inflammatory cell infiltration,and the degree of hepatocyte injury was related to the modeling time.Local capillary bile duct hyperplasia was occurred at 8 weeks,and capillary bile duct hyperplasia was evident at 12 weeks.Compared with the model group,the degree of cytoplasmic sparseness of hepatocytes in the YCHD group was reduced,the degree of bile duct reaction was reduced,and hepatocyte injury was alleviated.The high dose group was superior to the low dose group.The increased TC and PL level and decreased TBA level in bile result in a significant increase in CSI(P < 0.01).The expression levels of cholesterol metabolism-related genes(ABCG5,ABCG8,SRBI,CYP7A1,NPC1L1,etc.)in the liver and small intestine of the model mice were abnormal,and TC levels in liver tissue was significantly elevated(P<0.01).Compared with the model group,YCHD treatment could reduce the liver weight and the ratio of liver weight and body weight(P<0.05 or P<0.01).The serum levels of ALT,AST,ALP,TC and LDL-C were significantly reduced via YCHD(P<0.05 or P<0.01).The reduced level of TC and PL and increased level of TBA in bile via YCHD treatment could lead to a lower CSI(P<0.05 or P<0.01),and the condition of gallstones in mice was alleviated.The m RNA expressions of ABCG5,ABCG8,SRBI and NPC1L1 in liver and small intestine were decreased in different degrees after four-weeks and eight-weeks YCHD administration while the level of TC in liver tissue was also decreased,of which the high dose group had the best effect(P<0.01).Conclusion: Yinchenhao decoction can ameliorate serum and liver biochemical abnormalities of mice with cholesterol gallstones induced by lithogenic diet,adjust the balance of cholesterol,phospholipid and bile acid in the bile to improve biliary cholesterol supersaturation,and regulate cholesterol metabolism,that is,inhibit cholesterol absorption in the small intestine and affect the transport of cholesterol in the liver to prevent and treat cholesterol gallstones.