| Objective: To investigate the inhibitory effects of berberine on experimental gallstone formation and explore its machanism by comparing bile ingredients and morphologic changes of the mucosa after using berberine. We used rabbit models which were induced by high cholesterol diet(HCD) and certain group of which were interfered by berberine.Methods: Experimental rabbits groups A total of 30 Japanese rabbits of either sex(3-4 months old, 2.2-2.8kg)were divided into three groups at random: control (normal rabbits chow), lithogenic (1.2g high cholesterol diet) and berberine prevention (1.2g high cholesterol diet +100mg·d berberine) groups .These three groups were fed with formal diet for one week, following with the specific diet. After 8 weeks, 12-hour-fasted ainmals were killed.5ml bile in each rabbit was extracted, Cholecystectomy was performed. The bile was obtained for observing the crystal formation, The bile was judged glossly to determine whether there was sandy crystal(D＞1mm)and observed the colour, luster and viscidity, and keeped in -20℃icebox orderly. At the same time, the gallbladder wall and mucosa were also observed. The specimens from gallbladder were preserved in 10 % fonnalin, embedded in paraffin, sectioned onto mcroscopeslides at a thickness of 5um. Pathologic diagnosis was based on hematoxylin and cosin stain(H-E stain). check the level of TC, PL and BS. Blood was drawed separately to check the level of TG TC, LDL-C, HDL-C. Bile was obtained for observing gallstone formation, and the morphologic changes of the mucosa were assessed under microscopy.The level of COX-2 in gall-bladder mucosa in each group was estimated with immunohistochemical assay.Results: 1.Mucosa inflammation and bile cholesterol crystal formation: Gallbladder wall was thicker and paler in gallstone group than in berberine group. And the mucosa of gallstone group was seen congestion and edema, which was hardly seen in berberine. Cholesterol crystal formation was seen 3/10, 8/10 separately in berberine group and model group. The risk of cholesterol crystal formation was remarkably decreased in berberine group than in gallstone group(x~2 = 5.05 ) P＜0.05).2. Mucosa inflammation: Chronic inflammation was seen in two groups without obvious structure change. Inflammatory cells were noted in mucosa and submucosa, most of which was lymphocyte. Monocytes were only seen in submucosa layer. The inflammation level in berberine group was significantly lower than in gallstone group(P＜0.01 )3. The bile ingredients in high cholesterol diet group had significant changes compared to normal control group. The TC and CSI in gallstone group was significantly higher than the others(P＜0.05). CSI in each group was (control 0. 82±0. 23,lithogenic 1. 64±0. 27and berberine prevention 1. 13±0. 29). The BS in gallstone group was significantly lower than the other groups. TG, TC and LDL-C of Berberine group had an obvious difference compared with gallstone group (P＜0.05-0.01 )4.The degree of mucosa inflammation and level of COX-2 expression were significantly higher in gallstone group than in the other two groups (47.66±1.54vs28.39±1.46,P＜0.05 ;37.42±2.45vs47.66±1.54, P＜0.05) .Conclusions 1. COX-2 was highly expressed in gallbladder mucosa of gallstone during gallstone formation. Berberine may inhibiting the activity of COX-2.Bererine may reduce the extent gallbladder mucosa inflammatory, and decreased the cholesterol crystal formation in gallbladder.2. Bererine may decrease the risk of cholesterol gallstone formation by Regulate hyperlipidemia, reduced the quantity of biliary cholesterol and increase the quantity of chlic acid, which contributed to prevention of cholesterol gallstone formation.3. Chronic inflammatory is maybe concerned with the formation of cholesterol gallstone. Berberine can reduce the extent gallbladder mucosa inflammatory, and decreased the cholesterol crystal formation in gallbladder. |
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