Effect Of Danshiliuhao Decoction On Gallstone Formation And Mechanisms Research

Objective: To investigate the preventive and therapeutic effect of Danshiliuhaodecoction against cholesterol gallstone and on bile pigment stones, and explore thepossible mechanism.Methods:(1) Effect on C57mice of cholesterol gallstone Fifty mice were randomly divided intonormal group(n=10), untreated group(n=15), Danshiliuhao treated group(n=15) andTauroursodeoxycholic Acid treated group(n=10). Cholesterol gallstone was induced inmice of the latter three groups by feeding with high cholesterol diet. After2-month untreat-ment, mice of Danshiliuhao group and Tauroursodeoxycholic Acid group were intragastri-cly administered with Danshiliuhao decoction21g·kg-1·d-1and Tauroursodeoxycholic Acid12mg·kg-1·d-1.1-month later, animals were sacrificed and sampled to calculate the incide-nces of stone formation. The contents of serum cholesterols, SOD activity and MDA cont-ent in liver were detected.(2) The effects of guinea pig bearing bile pigment stones and Study on the mechanism ofresistance to bile pigment stone formation Forty guinea pigs were randomly dividedinto blank control group (n=10), model group (n=15) and Danshiliuhao treatedgroup(n=15). In addition to the blank control group, the other two groups were establishedwith diet-induced model of bile pigment stones. Simultaneously, Danshiliuhao treatedgroup was given7.7g·kg-1·d-1,ig for30days. The stone formation rate, serum bilirubinlevel, liver superoxide dismutase (SOD) activity, malondialdehyde (MDA) content weremeasured and CCKaR,5-HT3aR mRNA and protein expression were also analyzed at theend of diet treatment.Results: (1) Effect on C57mice of cholesterol gallstone The incidences of stone formation were85.71%for untreated group while it’s significantly higher in untreated group thanothers(P<0.05). Contents of serum LDL-C, AST, ALT, IBIL and TBA in the danshiliuhaogroup were lower than that in the untreated group.(P<0.05). Danshiliuhao decoction couldincrease the SOD activity and decrease the MDA content in liver tissues, showingsignificant differences as compared with those in the untreated group(P<0.05).(2) The effects of guinea pig bearing bile pigment stones and Study on the mechanism ofresistance to bile pigment stone formation One guinea pig that formed stone was found inblank control group, and the incidences of gallstone formation were10%, while thegallstone formation rate of model group was91.67%. Danshiliuhao decoction gallstoneformation rate was26.67%, which was significantly lower than that of model group. Thelevel of serum TBIL, DBIL and IBIL in the danshiliuhao group was lower than those in themodel group.(P<0.05). And the liver SOD activity was significantly higher in theDanshiliuhao decoction treatment group than the model group, while MDA content waslower than the model group. The results of RT-PCR indicated that expression level ofCCKaR,5-HT3aR of untreated group was significantly lower than the blank group, whilethe Danshiliuhao decoction treated group significantly higher than the blank group anduntreated group (P<0.05). Immunohistochemical result of5-HT3aR protein showed thatthe amount of5-HT3aR protein in untreated group was much lower than that in the blankgroup, while much higher in the Danshiliuhao decoction group than that in the untreatedgroup (P<0.05). But the immunohistochemical results showed that the amount of CCKaRprotein of untreated group was lower than that of the blank group, while higher in theDanshiliuhao decoction group than that in the untreated group, but the varience had nostatistical significance (P>0.05).Conclusion:(1) Danshiliuhao decoction can decrease the incidence of stone formation and improve thebiochemical parameters, which may be one of the mechanisms in the treatment and prevention of cholesterol gallstone disease.(2) Danshiliuhao decoction can reduce the gallstone formation of guinea pig bearing bilepigment stones, and its mechanism is, at least partly, due to the anti-oxidative stress.(3) Danshiliuhao decoction significantly affected the mRNA and protein expression ofCCKaR,5-HT3aR in guinea pig bearing bile pigment stones. Its therapeutic effect on bilepigment calculus achived partly by increasing the expression of CCKaR and5-HT3aR.