| PrefaceThe intestinal barrier function has drawn more and more attention in recent years for its important role in many diseases. More and more diseases were found influenced by intestinal barrier during their development. In our preliminary experiments about the pigment gallstone, we found some signs of the intestinal barrier function disorder during the pigment gallstone formation which may suggest the potential role in its pathogenesis. Are the immunologic barrier and biologic barrier of intestinal mucosa destroyed in this course? Do they play a significant role in this course? This study was undertaken in guinea pig model to investigate the possible action and mechanism of the intestinal immunologic barrier function in the pigment gallstone pathogenesis. The experiment was arranged though a clue: intestinal barrier injury—bacteria translocation—enzyme in bile change—gallstone formation. A series of indexes about the intestinal immunologic barrier and morphology of intestine mucosa were observed such as:LPL(Lamnina Propria Lymphocytes),IEL(intraepithelial lymahocyte),sIgA-Plasmcyte,T lymph-cell and B lymph-cell, 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 barrier.MethodsEighty guinea pigs (weighing 200~250g) were divided randomly into normal group (CON), pigment gallstone group (PS) and intestinal mucosa protection group (GLN). 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 collected for detection after eight weeks. The gallstone-forming rate, morphology of intestinal mucosa, the cell populations of LPL per high power lens, the cell populations of IEL per high power lens, the cell populations of CD3~+ T-cell per high power lens, the cell populations of CD40~+ T-cell per high power lens, the cell populations of slgA-Plasmcyte per high power lens, 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 10% neutral formalin, paraffin imbeded. Pathology examination: the pathology examination of intestinal mucosa were performed by HE stain, to count the cell populations of LPL per high power lens, the cell populations of IEL per high power lens. To count the population of gut mucsosal lymphocyte by immunohistochemistry: the ileum tissues 5cm away from the cecum dealed with immunohistochemistry, be tested under microscope, the buffy cell is the masculine cell, then counting the cell populations of CD3~+ T-cell per high power lens, the cell populations of CD40~+ T-cell per high power lens, the cell populations of sIgA-Plasmcyte per high power lens Data are expressed as mean±SD. All analyses were undertaken using t analysis by SPSS 11.5 software. Ap value of≤0.05 was accepted as the level of statistical significance.ResultsThe incidence of Gallstone formation was 68.4% 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 (36.8%) compare with that in PS group (p<0.05).Compare with that of CON group, the microstructure of intestinal mucosa cell in PS group showed some signs of disorder: the villus of the intestinal mucosa in a disorder way, intestinal mucosa is dropsical, there were inflammatory cell infiltrate and hemangiectasia in the intertinal mucusa. Layers of intestinal wall became thinner, the height of intestina villus depressed, the width of intestina villus dinished; In GLN group, the villus of the intestinal mucosa in a disorder way, intestinal mucosa is dropsical, there were inflammatory cell infiltrate and hemangiectasia in the intertinal mucusa, but the situation is less serious.Compare with the CON group, both the cell populations of LPL per high power lens and the cell populations of IEL per high power lens in PS group are less (p<0.05, p<0.01) . While in GLN group the numerical value of cell population were higher than that in PS group, tab 1.To count the population of gut mucsosal lymphocyte by immunohistochemistry: counting the cell populations of CD3~+ T-cell per high power lens. Compare with the CON group In PS group and GLN group the numerical value of masculine cell population is lower, While in GLN group the numerical value of masculine cell population were higher than that in PS group, tab 2. Counting the cell populations of CD40~+ B-cell per high power lens. Compare with the CON group In PS group and GLN group the numerical value of masculine cell population is lower, While in GLN group the numerical value of masculine cell population were higher than that in PS group, tab 2.Counting the population of the cell populations of sIgA-Plasmcyte by immunohistochemistry: counting the cell populations of sIgA-Plasmcyte per high power lens. Compare with the CON group In PS group and GLN group the numerical value of masculine cell population is lower, While in GLN group the numerical value of masculine cell population were higher than that in PS group, tab 2. ConclusionsIntestinal mucosa immunologic barrier function and mechanical barrier function disorder can be found in the process of pigment gallstone formation in diet-induced pigment gallstone guinea pig model. The intestinal barrier is correlated with the pathogenesis of pigment gallstone. Glutamine can improve the intestinal barrier function and decrease the rate of pigment gallstone formation. The intestinal barrier function disorder may take part in the process of promoting pigment gallstone formation through bacteria translocation, endotoxemia and affecting the activity of biliaryβ-glucuronidase. In this study we pay attention to the role of Intestinal mucosa immunologic barrier function and mechanical barrier function disorder in the process of pigment gallstone formation in diet-induced pigment gallstone guinea pig model, and presume its mechanism preliminarily, but the specific relation need to be researched and studied.
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