To Explore The Mechanism And Application Of Dahuanglinxian Capsule In The Prevention Of Gallstones Based On Metabonomics

Objective: Observation the intervention treatment efficacy and bile characteristics of metabonomics of Da Huang Lingxian capsules to gallstone patients,and discuss the mechanism of action in prevention and control of of gallstones in C57 BL / 6 mice model.Methods:(1)Clinical research: 142 patients were clearly diagnosed as gallstones and chronic cholecystitis were randomized divided into Da Huang Lingxian capsules group 72 cases and anti-inflammatory cholagogue control group 70 cases.In addition to the other normal medication treatment,intervention group given oral Da Huang Lingxian capsules 3 times a day,6 pills;Control group given oral anti-inflammatory cholagogue 3 times a day,6 pills,treatment for 3 months.Statistics of two groups patients before and after the treatment of TCM symptom score,gallbladder inflammation and the situation of gallstones improve integral under type-B ultrasonic,bile endogenous metabolites characteristics change(GC-MS),and observed the blood laboratory testing after treatment,follow-up evaluation of its safety and efficacy.(2)Fundamental research: 50 male C57BL/6 mice according to the statistical software were randomly divided into normal control group(N group),model group(M),ursodeoxycholic acid control group(U)and normal drug control group(Y),Da Huang Lingxian capsules treatment group(group D),each group have 10 mice.Besides N and Y group were given with normal feed,M,U,D group were given a high-fat, high cholesterol(15% fat,1% cholesterol,0.5% cholic acid)to feed.According to the equivalent dose conversion and adult body weight in mice,U group was lavaged with 130 mg/kg?d UDCA Dry powder solution,group Y and D were lavaged with 13 g/kg?d Da Huang Lingxian capsules liquid respectively during the period of building model.N and M group were respectively given the same volume of physiological saline lavage.8 weeks later,collection in mice serum biochemical examination,gallstones testing its stones ingredients,take the same parts of the liver tissue,respectively using immunohistochemical,PCR and Western blotting technology to detect the hepatic duct side form cell membrane protein gene and protein expression differences.Results:(1)clinical research:(1)Compared with the control group,intervention group curative effect is slightly higher(P=0.04),the improvement of the symptoms integral is significantly higher(P<0.001),and the improvement within the gallbladder stones is better(P=0.03).Besides these,bad bile acoustic,wall coarse improvement after treatment is better than the control group(P=0.04,P=0.04).(2)Comparison before and after the treatment,two groups of patients with blood test indexes are improved,but the improvement with WBC,ALT,AST,TBIL,DBIL in intervention group were bigger compared with control group(P<0.05),respectively.(3)In the two groups before and after treatment in patients with biliary endogenous metabolites except the LDL,other metabolites were significant changed(P<0.001),and compared with control group,intervention group alanine,citric acid,lactic acid,glycine,cholesterol,glycerin,malic acid,choline,bezoar,alkali metabolites concentration change difference had statistical significance(P<0.05).(4)Two groups patients were not apparent serious adverse reactions,and special events. (2)Fundamental research:(1)Group N,Y have no gallstones,M group mice into gallstones at a rate of 100%,within the gallbladder,sediment precipitation and grain appearance can be seen.Group U and D stone rate is 44.44%,30% respectively,the two groups rate were lower than M group(P<0.0125).(2)Useing potassium bromide tabletting method of infrared spectrum,we analysis the gallstones in mice is the cholesterol stone.(3)PCR results: compared with group M,group N,Y,D mice of Abcb11 and Abcc2 were relatively higher(P<0.001).Compared with U group,group D mice Abcb11 m RNA expression is higher(P<0.001).And group N,U Abcc2 m RNA expression were higher compared with group D(P=0.002).(4)Immunohistochemical results: compared with group M,group N,U,Y,D of Abcb11 and Abcc2 protein expression in mice were increased(P< 0.001),and among the other four groups,there were no significant differences compared with each other.(5)Western blotting results: compared with group M,group N,U,Y,D of Abcb11 and Abcc2 protein expression in mice were increased(P<0.001).Compared with the N group,group D Abcc2 protein expression was dropped(P=0.002),but no significant difference compared with U group.Conclusion: Da Huang Lingxian capsules can improve patients with gallstones in biliary metabonomics characteristics,serum biochemical indicators and gallbladder inflammation,at the same time can improve the gallbladder stones in mice of Abcb11 and Abcc2 transport protein gene and protein expression,make it close to normal physiological level,improve the secretion of pathologic bile and bile composition,so as to achieve the effect of the prevention and control stone.In addition,Da Huang Lingxian capsules will not cause some special adverse reactions and special side effects.