| Objective:Observe the expression of interleukin-17(IL-17) and fork transcription factor3(Foxp3) in liver and small intestine in autoimmune hepatitis (AIH) mice, and on this basis to discuss the relationship between AIH mice liver inflammation and intestinal immunologic injury and the role of T help cell type17(Th17) and CD4+CD25+regulatory T cells (Tregs) in the development of AIH.Methods:1. Model building female Balb/c mice,6-7weeks, a total of12, were randomly divided into experimental group (6) and the control group (6). Experimental mice by caudal vein injection Concanavalin A (Con A)12.5mg/kg, once a week,4weeks continuous injection; Does not make processing control group, two groups under the same breeding environment.2. Kill all of the mice after4weeks and take its liver and intestinal tissue samples frozen, using RT-PCR to test IL-17and Foxp3expression in liver and small intestine; Liver and intestinal tissue fixed by10%formaldehyde,conventional embedding, sectioning after hematoxylin-eosin (HE) staining pathologic examination and immunohistochemical staining detection IL-17, Foxp3expression.3. The comparative analysis of the experimental group and control group in the liver and intestinal tissue IL-17,Foxp3expression, compared two groups of mice liver and intestinal inflammation,IL-17Foxp3and its role on the pathogenesis of AIH.4. SPSS18.0statistical software was used to all materials statistical analysis, with P<0.05for the difference was statistically significant.Results:1. Successfully established AIH experimental animal models in mice. Experimental mice liver HE staining showed that the liver tissue inflammation mainly for a large number of lymphocytes and a small amount of neutrophil infiltration, hepatic lobule structure obvious damage, liver cell edema, ballooning, dot necrosis or even fragmented necrosis, etc; While the control group normal liver tissue structure.2. According to the results of RT-PCR, mice of experimental group IL-17mRNA gene expression in liver is significantly higher than the control group (3.79±0.90VS1.00±0.00), the difference was statistically significant (P<0.05); Experimental Foxp3mRNA gene expression in liver tissue of mice significantly lower than the control group (0.13±0.03VS1.00±0.00), the difference was statistically significant (P<0.05);3. According to the results of RT-PCR, mice of experimental group IL-17mRNA gene gene expression in the intestinal tissue is significantly higher than the control group (4.32±1.00VS1.00±0.00), the difference was statistically significant (P<0.05); Experimental mouse small intestine tissue Foxp3mRNA gene expression was lower than that of control group (0.21±0.13VS1.00±0.00), the difference was statistically significant (P<0.05).4. The pathological tissue slice staining, found that the experimental group small intestine histopathologic grading of moderately severe inflammation, control group no to mild inflammation.5. Immunohistochemical staining showed that the experimental group in mice liver and intestinal tissue in the expression of IL-17raised obviously, and the control group in the liver and intestinal tissue no or only a small amount of IL-17express (liver4.83±1.17VS0.33±0.52;intestine6.83±1.17VS0.67±0.52);In the control group mice liver and intestinal tissue is scattered distribution of Foxp3expression, and the experimental mice liver tissue and although there are a large number of lymphocyte infiltration in the small intestine tissue, but Foxp3expression is less than that of control group, and dyed lighter (liver0.50±0.84VS1.83±0.75;intestine1.17±0.75VS5.17±1.47)6. The correlation analysis results showed that IL-17mRNA in the liver and intestinal tissue expression was positively correlated (r=0.903, P<0.05); Foxp3mRNA expression in liver and intestinal tissue was positively correlated(r=0.973, P<0.05).Conclusions:1. IL-17expression in liver tissue in AIH mice increased significantly, prompt Th17raised; Foxp3expression in liver tissue in AIH mice significantly reduced, prompt Tregs downgraded.2. IL-17expression in intestinal tissue in AIH mice increased significantly, prompt Th17raised; Foxp3expression in intestinal tissue in AIH mice significantly reduced, prompt Tregs downgraded.3.Th17rise trend in AIH mice liver and intestinal tissue is consistent, Tregs cut trend in AIH mice liver and intestinal tissue is consistent, prompt there is some relationship between the immune liver injury and intestinal immune injury, but the specific mechanism is unclear.4. Th17and Tregs express imbalances may be an important factor for the liver and intestinal immune inflammatory lesions, and it plays an important role in the process of development of AIH, rebuild the balance may delay the progress of AIH. |
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