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目的探讨肠三叶因子(ITF)对小鼠炎症性肠病(IBD)肠上皮细胞及脾淋巴细胞凋亡的调节作用,分析ITF对IBD的保护作用机制。方法 48只小鼠随机分为3组(n=16):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模型后每只小鼠给以rITF 0.1 mL腹腔注射;乙醇对照组,未造模型小鼠每只给予9 g.L-1盐水0.1 mL腹腔注射。各组均连续5 d注射,第6天评估疾病活动度指数(DAI),处死动物后取其结肠组织及脾脏组织,测定其结肠组织髓过氧化物酶(MPO)活性、应用原位末端标记试剂盒及流式细胞技术检测其肠上皮细胞、脾淋巴细胞凋亡情况及脾淋巴细胞Fas表达情况。结果与TNBS组小鼠相比,rITF组小鼠在死亡只数、大便性状、血便情况、体质量下降情况等临床症状及肠道局部炎症均明显减轻,MPO值显著降低(P<0.05),肠上皮细胞凋亡指数显著下降(P<0.01);乙醇对照组IBD小鼠脾淋巴细胞的凋亡明显不足,且Fas表达下调,但TNBS组与rITF组间凋亡率无显著性差异。结论 ITF对IBD小鼠具有保护作用,并且可能通过调节IBD小鼠肠上皮细胞凋亡水平保护肠上皮细胞,但ITF对脾淋巴细胞的凋亡无调节作用。
Objective To investigate the regulatory effect of intestinal trefoil factor (ITF) on the apoptosis of intestinal epithelial cells and splenic lymphocytes in mice with inflammatory bowel disease (IBD) and to analyze the protective mechanism of ITF on IBD. Methods 48 mice were randomly divided into three groups (n = 16): TNBS group. After establishing IBD model by TNBS, each mouse was injected intraperitoneally with 0.1 mL of 9 g / L saline; The intestinal trefoil factor (rITF) group was injected intraperitoneally with rITF 0.1 mL after IBD model was established. The mice in the ethanol control group and the non-model mice were intraperitoneally injected with 0.1 mL of 9 gL-1 saline. The rats in each group were injected continuously for 5 days and the disease activity index (DAI) was evaluated on the 6th day. The colon and spleen tissues were harvested and the activity of myeloperoxidase (MPO) in colon tissues was determined. Kit and flow cytometry to detect the apoptosis of intestinal epithelial cells and splenic lymphocytes and the expression of Fas in splenic lymphocytes. Results Compared with TNBS group, the clinical symptoms and intestinal inflammation in rITF group were significantly reduced and the MPO value was significantly decreased (P <0.05) in the number of death, stool, bloody stools and body weight, The apoptosis index of intestinal epithelial cells was significantly decreased (P <0.01). The apoptosis of splenic lymphocytes in IBD mice in ethanol control group was obviously insufficient and the expression of Fas was down-regulated. However, there was no significant difference in apoptotic rate between TNBS and rITF groups. Conclusion ITF has a protective effect on IBD mice and may protect intestinal epithelial cells by regulating intestinal epithelial cell apoptosis in IBD mice, but ITF has no effect on the apoptosis of splenic lymphocytes.