| Objective:To prepare gallbladder cholesterol stone guinea pig model by high cholesterol-induced stone feed induction method,and to intervene with Chaihu Shugan Powder,Longdan Xiegan Decoction and Chaihu Shugan Powder and Longdan Xiegan Decoction.By observing the formation of stones after intervention and the morphology of the liver,serum total cholesterol(TC),triglyceride(TG),and low-density lipoprotein cholesterol(LDL-C),cholesterol(T-CHO),total bile acid(TBA)content,and determination of cholestrol 7-alpha hydroxyl-lase(CYP7A1),bile salt output pump(BSEP)mRNA expression.The effects of soothing liver and qi,eliminating heat and dampness on blood lipids and liver cholesterol metabolism related genes in gallbladder cholesterol stone guinea pigs,and exploring the mechanism of the treatment of gallbladder cholesterol stones by soothing liver and qi,eliminating heat and dampness.Methods: 50 normal guinea pigs,half male and half female,were randomly divided into blank group(n=8)and experimental group(n=42).The blank group was fed with normal diet.The experimental group was fed with high cholesterol to feed the stone to prepare the gallbladder cholesterol stone model.After 8 weeks of modeling,the experimental group randomly selected 10 samples to test whether the modeling was successful.After successful modeling,the experimental group of guinea pigs were randomly divided into model group,Chaihu Shugan San group,Longdan Xiegan Decoction group and combined medication group,8 rats in each group,and were continuously administered for 4 weeks.Blank group and model group were given normal saline and drug intervention groups were given Chaihu Shugan Powder,Longdan Xiegan Decoction,Chaihu Shugan Powder and Longdan Xiegan Decoction.After the last administration for 24 hours,the body weight of each group of guinea pigs was weighed.After anesthesia with chloral hydrate,blood was collected from the heart,and liver and gallbladder tissues were harvested to observe the formation of cholesterol gallstones in the gallbladder.Observing the histomorphology of the liver by the method of hematoxylin-eosin staining;measuring the content of TC and TG by end point method;measuring the content of LDL-C by elimination method;measuring the T-CHO content by single reagent colorimetry;measuring the TBA content by enzyme colorimetry;and measuring the expression levels of CYP7A1 and BSEP mRNA in liver by real-time fluorescence quantitative polymerase chain reaction.Results: 1.Body weight comparison: Compared with the blank group,the body weight of the model group,Chaihu Shugan San group,Longdan Xiegan Decoction group and the combination group had significant decrease,and there was statistical difference(P<0.01).Compared with the model group,the weight of guinea pigs in Chaihu Shugan San group,Longdan Xiegan Decoction group and combination group increased,and there was statistical difference(P<0.05).There was no significant difference in body weight between the guinea pigs in each group(P>0.05).2.Comparison of stone formation: The blank group,model group,Chaihu Shugan San group,Longdan Xiegan Decoction group,combined drug composition stone rate were 0,100%,25.0%,37.5%,50.0%.3.Liver H-E staining showed that the guinea pig liver cells in the model group had obvious fatty degeneration,accompanied by varying degrees of inflammatory cell infiltration,cytoplasm loose,and containing large fat droplets;Drug intervention in each group of guinea pig liver cell steatosis and inflammation and necrosis were significantly improved;The liver cells in the blank group were normal in morphology,and no lipid droplets accumulated in the cytoplasm.4.Comparison of serum TC,TG,LDL-C content: Compared with the blank group,the serum levels of TC,TG and LDL-C in the model group were significantly increased(P<0.01);Compared with the model group,the serum levels of TC,TG and LDL-C in Chaihu Shugan San and Longdan Xiegan Decoction group were significantly lower(P<0.05);Compared with the model group,the serum levels of TC in the combination group were significantly lower(P<0.05);Compared with the model group,serum LDL-C levels in the combination group were significantly lower(P<0.05);There was no significant difference in drug intervention between the three groups(P>0.05).5.Comparison of T-CHO and TBA content in bile: Compared with the blank group,the T-CHO content of the bile in the model group was significantly increased,and the TBA content was significantly decreased,with statistical difference(P<0.01);Compared with the model group,the content of bile T-CHO in Chaihu Shugan San group,Longdan Xiegan Decoction group and combination group was significantly lower,with statistical difference(P<0.01);Compared with the model group,the content of bile TBA in Chaihu Shugan Powder group was significantly increased,with statistical difference(P<0.01),and the content of bile TBA in Longdan Xiegan Decoction group and combined drug group was increased,with statistical difference(P<0.05);There was no significant difference in bile T-CHO and TBA between the three groups(P>0.05).6.Comparison of liver CYP7A1 and BSEP mRNA expression levels: Compared with the blank group,the expression of CYP7A1 and BSEP mRNA in the model group was significantly lower(P<0.01);Compared with the model group,the expression of CYP7A1 and BSEP mRNA in the liver of Chaihu Shugan Powder group was significantly increased(P<0.01);Compared with the model group,the expression of CYP7A1 mRNA in the liver of Longdan Xiegan Decoction group was significantly increased(P<0.01);Compared with the model group,the expression of BSEP mRNA in the liver of Longdan Xiegan Decoction group and the combination group was increased,with statistical difference(P<0.05);Compared with the model group,the expression of BSEP mRNA in the liver of guinea pigs in the combination group was increased,with no significant difference(P>0.05);There was no significant difference in the expression of CYP7A1 and BSEP mRNA between the three groups(P>0.05).Conclusion: 1.Soothing liver and qi,eliminating heat and dampness method can play its role in choleretic discharge by lowering blood lipid concentration and improving cholesterol metabolism.2.Soothing liver and qi,eliminating heat and dampness can increase its expression of CYP7A1 and BSEP mRNA and improve cholesterol metabolism to exert its choleretic effect. |
