| Objective: C57BL/6 mice were used as the research object to establish a gallbladder stone model.Western Blot and RT-PCR were used to compare the expression changes of CYP7A1 and CYP8B1 genes on the lateral membrane cells of the hepatobiliary duct,and the intervention effect of Dahuang Lingxian capsule.From the central link,cell secretion state,the mechanism of gallstone formation was further explored from the central link of cell secretion state,and the target of Dahuang Lingxian capsule in the prevention and treatment of gallstone was further explored.Methods:(1)Group: 60 adult SPF male C57 BL / 6 mice were randomly divided into five groups: group P(blank control group),group Z(lithogenic group),group W(irrigation group),group U(ursodeoxycholic acid capsule group)and group D(Dahuang Lingxian capsule group),with 12 mice in each group.Mice were fed with high-fat,high calorie and high cholesterol diet(15% fat,1.5% cholesterol and 0.5% cholic acid)to induce gallstone formation to establish gallstone model.Water and drugs were given by gavage during the modeling process.(2)Specimen collection: The mice were dissected and the gallbladder was taken to observe the model,and the liver tissue samples were kept for the detection of cytokines.(3)Detection: Western Blot and RT-PCR were used to detect the transcription levels and the changes in protein expression of CYP7A1 and CYP8B1 of hepatobiliary duct lateral membrane cells in each group.Results:(1)Modeling: there was no gallstone formation in group P(blank control group),and the bile was light yellow and crystal clear.There were granular or silt-like stones in the gallbladder in group Z(stone-causing group)and group W(irrigation group).Among them,11 cases of group Z formed stones,with a stone formation rate of 91.7%,and 10 cases of group W(irrigation group)formed stones,with a stone formation rate of 90.9%.The stone formation rate of group U(ursodeoxycholic acid capsule group)and group D(Dahuang Lingxian capsule group)decreased significantly.The bile in the gallbladder that did not form stones was observed to be pale yellow and crystal clear.There were 2 cases of stone formation in group U(16.7%)and 3 cases in Group D(25.0%).The results showed that after statistical analysis,comparing group D with group W,the difference between the two groups was statistically significant(P<0.01),but ompared with group U,the difference between the two groups was not statistically significant(P>0.05).It can be considered that Dahuang Lingxian capsule has the effect of reducing the formation of gallstone,that is,preventing the formation of gallstones,and its effect is equivalent to that of ursodeoxycholic acid capsules.(2)m RNA transcription levels of CYP7A1 and CYP8B1 in hepatobiliary lateral membrane cells of mice in each group: Real-Time PCR technology was used to detect the m RNA expression levels of CYP7A1 and CYP8B1 in each group of mouse hepatobiliary membrane cells.Compared with group W,the difference in CYP7A1 and CYP8B1 m RNA expression levels in group D was statistically significant(P<0.01),and the expression levels of CYP7A1 and CYP8B1 m RNA in group D were significantly higher than those in group W.Compared with group U,there was no significant difference in the expression levels of CYP7A1 and CYP8B1 m RNA in group D(P>0.05).(3)Protein expression levels of CYP7A1 and Cyp8b1 in hepatobiliary lateral membrane cells of mice in each group : The Western-Bloting method was used to detect the protein expression levels of CYP7A1 and CYP8B1 in each group of in hepatobiliary lateral membrane cells of mice.Compared with group W,the difference in protein expression of CYP7A1 and CYP8B1 in group D was statistically significant(P<0.01).And the CYP7A1 and CYP8B1 protein expression levels in group D were significantly higher than those in group W.Compared with group U,the differences in CYP7A1 and CYP8B1 protein expression levels in group D were not statistically significant(P>0.05).Conclusion:(1)Dahuang Lingxian capsule can prevent and treat the formation of gallbladder stones in mouse models,and its effect is equivalent to that of ursodeoxycholic acid capsule.(2)Dahuang Lingxian capsule can significantly increase the m RNA expression levels of CYP7A1 and CYP8B1 in mice hepatobiliary lateral membrane cells,make the FXR-CYP7A1/CYP8B1 receptor system adjust to a normal physiological state,and avoid cholestasis and pathological bile production to achieve the effect of reducing the occurrence of gallstones.(3)Dahuang Lingxian capsule can stabilize the protein expression levels of CYP7A1 and CYP8B1,make the FXR-CYP7A1/CYP8B1 receptor system adjust to a normal physiological state,and avoid cholestasis and pathological bile production to achieve the effect of reducing the occurrence of gallstones. |
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