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目的探讨肠三叶因子(ITF)对小鼠炎症性肠病(IBD)肠道病变的影响,分析ITF对小鼠IBD肠道先天性免疫功能的调节作用。方法 48只小鼠随机分为3组,每组16只。三硝基苯磺酸(TNBS)组:用TNBS建立IBD模型后每只小鼠给予9 g.L-1盐水0.1 mL腹腔注射;基因重组肠三叶因子(rITF)组:建立IBD模型后每只小鼠给予rITF 0.1 mL腹腔注射;乙醇对照组:未造模型小鼠每只小鼠给予9 g.L-1盐水0.1 mL腹腔注射。各组均连续5 d注射,第14天评估小鼠临床表现,麻醉处死小鼠后取其结肠组织作组织学评分,HE染色行肠组织病理学检查,并采用免疫组织化学法及实时定量PCR法检测其肠道TNF-α及Toll样受体(TLR4)和核因子(NF-κBp65)基因、蛋白表达水平。结果与TNBS组小鼠比较,rITF组小鼠死亡只数、大便性状、血便情况、体质量下降情况等临床表现及肠道局部炎症均明显减轻,肠道病理大体观及组织学评分均明显下降(Pa<0.05),TNF-α蛋白表达水平明显下降(Pa<0.05),TLR4、NF-κBp65基因及蛋白表达明显下调(Pa<0.01)。乙醇对照组无死亡,临床症状及肠道局部炎症轻微,TNBS组TNF-α、TLR4、NF-κBp65基因及蛋白表达明显高于乙醇对照组(Pa<0.01)。结论 ITF对IBD小鼠肠道病变具有保护作用,可能与调节肠道先天免疫有关。
Objective To investigate the effect of intestinal trefoil factor (ITF) on intestinal pathology in mice with inflammatory bowel disease (IBD) and to study the regulatory effect of ITF on the intestinal tract innate immune function in mice. Methods 48 mice were randomly divided into three groups of 16 rats. Trinitrobenzene sulfonic acid (TNBS) group: After IBD model was established by TNBS, each mouse was injected intraperitoneally with 0.9 g / L saline 0.1 mL; Recombinant intestinal trefoil factor (rITF) group: Rats were given rITF 0.1 mL intraperitoneal injection; ethanol control group: mice without model were given intraperitoneal injection of 0.1 mL of 9 gL-1 saline. The rats in each group were injected continuously for 5 days. The clinical manifestations of mice were evaluated on the 14th day. The colon tissues were harvested for histological evaluation after anesthetized mice were killed. The histopathological examination was performed by HE staining. Immunohistochemistry and real- Methods The mRNA and protein expressions of TNF-α, TLR4 and NF-κB p65 in the intestine were measured. Results Compared with TNBS group, the clinical manifestations and intestinal inflammation in rITF group were significantly decreased as the number of death, stool, bloody stool and body weight were decreased, and gross pathological gross and histological scores were significantly decreased (P <0.05). The expression of TNF-αprotein was significantly decreased (P <0.05), while the expression of TLR4 and NF-κBp65 was downregulated (P <0.01). The ethanol control group had no death, clinical symptoms and slight intestinal inflammation. The expressions of TNF-α, TLR4 and NF-κBp65 in TNBS group were significantly higher than those in ethanol group (P <0.01). Conclusion ITF has a protective effect on intestinal lesions in IBD mice, which may be related to the regulation of intestinal innate immunity.