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目的通过研究肠三叶因子(ITF)对新生大鼠坏死性小肠结肠炎(NEC)模型肠组织病理学改变及肠道组织中蛋白酶Caspase-3、蛋白Bax和Bcl-2的含量变化,探讨ITF对NEC保护作用的可能机制。方法30只新生1日龄Wistar大鼠随机分为3组,正常对照组、实验组、干预组,每组10只。实验组为NEC模型后加生理盐水0.2ml腹腔注射;干预组为NEC模型后予以腹腔注射ITF 0.2 mg(0.2 ml);正常对照组未予处理。第4天处死所有大鼠,取肠组织待检,取近回盲部1～2 cm肠道组织,采用分光光度法检查Caspase-3的表达、采用免疫组化法检测肠道组织中Bax及Bcl-2的含量变化并做病理学检查。结果实验组Caspase-3表达高于正常对照组和干预组(P均<0.05),干预组与正常对照组比较差异无统计学意义(P>0.05);实验组Bax表达高于正常对照组和干预组(P均<0.05),干预组与正常对照组相近(P>0.05);干预组Bcl-2表达高于正常对照组和干预组(P均<0.05),实验组高于正常对照组(P<0.05)。正常对照组的肠组织病理学未见异常,病理评分为0分;实验组HE染色切片见肠壁损伤轻重不一,可见全肠黏膜绒毛坏死,病理评分的中位积分为3分;干预组肠上皮细胞少量脱落,顶端绒毛坏死,病理评分的中位积分为1分。与实验组比较,ITF治疗后NEC导致的组织病理学改变明显减轻。结论注射ITF可通过减少Caspase-3、Bax表达和增加Bcl-2表达减轻NEC肠道损伤。
Objective To study the intestinal histopathological changes and the changes of protease Caspase-3, Bax and Bcl-2 levels in neonatal rats with necrotizing enterocolitis (NEC) by studying the effect of intestinal trefoil factor (ITF) Possible mechanism for NEC protection. Methods Thirty newborn Wistar rats of 1 day old were randomly divided into 3 groups: normal control group, experimental group and intervention group, with 10 rats in each group. The experimental group was treated with intraperitoneal injection of 0.2 ml saline (0.2 ml). The rats in the intervention group received intraperitoneal injection of 0.2 mg ITF (0.2 ml). The rats in the normal control group were not treated. On day 4, all the rats were sacrificed and the intestine tissue was taken for examination. The intestinal tissues of 1-2 cm proximal to the ileocecal region were obtained. The expression of Caspase-3 was detected by spectrophotometry. The expressions of Bax and Bcl-2 levels change and do pathological examination. Results The expression of Caspase-3 in the experimental group was higher than that in the normal control group and the intervention group (P <0.05), but there was no significant difference between the intervention group and the normal control group (P> 0.05) (P <0.05). The expression of Bcl-2 in the intervention group was significantly higher than that in the normal control group and the intervention group (P <0.05), and the level in the experimental group was higher than that in the normal control group (P <0.05). The histopathology of the normal control group showed no abnormality and the pathological score was 0; in the experimental group, the damage of intestinal wall was observed with HE staining, the total intestinal mucosa villus necrosis was observed, and the median score of pathological score was 3; A small amount of intestinal epithelial cells shedding, the top of the villous necrosis, the pathological score of the median score of 1 point. Compared with the experimental group, the histopathological changes induced by NEC after ITF treatment were significantly reduced. Conclusion Injecting ITF can reduce intestinal damage of NEC by decreasing Caspase-3, Bax expression and increasing Bcl-2 expression.