The Study On The Relationship Between The Intestinal Mucosal Barrier And The Pigment Gallstone In Guinea Pig Model

PrefaceThe cholelithiasis is the one of common ailment and frequently occurring illness in the world now. Cholesterol calculus developing in gallbladder had been cured by operation; Bile pigment calculus develop in intrahepatic bile ductour (90%)in our clinical practice in our country. Bile pigment calculus display the high rate of calculus remain, recidivation, operative concurrent, reoperation. Then, it has actual significance to research pathogenetic reason and precaution. In 1950s,it was reported the dependability of bile pigment calculus and bacterial infection with refractoriness. The high bacterial detective rate in bile in cholelithiasis patient cue the intimate relationship of bacterial infection with bile pigment calculus. But the resource of the bacterium is not definitive. There are the resource of the blood,gut, bacterial transplace. It is possible concomitance meanwhile. In our preliminary experiments about the pigment gallstone, we found some signs of the intestinal mechanical dysfunction during the pigment gallstone formation which may suggest the potential role in its pathogenesis. This study was undertaken in guinea pig model and cholelithiasis patients to investigate the possible action and mechanism of the intestinal barrier function in the pigment gallstone pathogenesis. The experiment was arranged though a clue: respecting the especial anatomy in the portal vein,when it happened abnormality in intestinal barrier,whether the bacterium permeate the gut by the something factor, through the hub of blood and lymph, soak hepatobiliary system, forming the bile pigment calculus. A series of indexes about the intestinal mucosal barrier were observed such as: bacterial classify cultivation, immunohistochemistry, et al. Glutamine was used as an intestine mucosa protector. We hope to investigate the pathogenesis of pigment gallstone in our experiment though a new angle: intestinal mucosal barrier.MethodsNinety-five guinea pigs (weighing 210～250g) were divided randomly into normal group (CON group,n=15), pigment gallstone group (PS group,n=30),short term pigment gallstone group (STPS group,n=20) and intestinal mucosa protection group (GLN group,n=30). Normal forage, pigment gallstone-forming forage and pigment gallstone-forming forage with supplemental intestinal mucosa protector (glutamine) were given to each group. The intestine tissue and intestinal contents were collected for detection after eight or four weeks. The gallstone-forming rate, morphology of intestinal mucosa, intestinal contents bacterial classify cultivation, intestinal mucosal immunohistochemistry were detected.In animal group, the gallstone can be identified easily during operation and the categories of the gallstones were confirmed by infrared spectrometry. The ileum tissues 5cm away from the cecum was isolated and immediately fixed in 30g/L glutaraldehyde for 24-hour and then dehydrated and dyed according to the standard sample preparation procedure for immunohistochemistry. The intestinal contents were attenuated to bacterial classify cultivation. The quantity of Bacillus bifidus ang Enteric bacilli ratio is the colonization resistance.ResultsThe incidence of Gallstone formation was 68.42% in PS group suggesting that the gallstone-inducing animal model was successfully established. While the incidence of gallstone formation in GLN group in which glutamine was used to protect the intestinal barrier was significantly decreased (38.89%), the incidence of Gallstone formation was 36.36% in STPS group compare with that in PS group (p<0.05).Compare with that of CON group, In PS group the bacterial classify cultivation of intestinal contents had some signs of disorder: the high of the Enteric bacilli and Enterococcus(p<0.05),the low of Bacillus bifidus and Bacterium lacticum(p<0.05). In STPS group showed: the high of the Enteric bacilli(p<0.05), the low of Bacillus bifidus(p<0.05). In GLN group showed: the low of Bacillus bifidus(p<0.05). In PS group, colonization resistance is degrade(p<0.05). The bacterial barrier in the gut was descended.Immunohistochemistry displayed that compare with that of CON group, the quantity of DC cell was increasing in STPS group(p<0.05). The quantity of B cell, plasmacytoid lymphocyte,SIgA was decreasing in PS group(p<0.05). The immunizing barrier in the gut was descended.ConclusionsThe intestinal barrier is correlated with the pathogenesis of pigment gallstone. Glutamine can decrease the rate of pigment gallstone formation. There was abnormality of permeability of the intestinal mucosa in the cholelithiasis patient.The intestinal barrier function disorder may take part in the process of promoting pigment gallstone formation.