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目的探讨肠三叶因子(intestinaltrefoilfactor,ITF)对坏死性小肠结肠炎(necrotizingenterocolitis,NEC)新生大鼠的肠黏膜组织中iNOS、TNF-α、NO的含量的影响,及ITF对NEC是否具有保护作用。方法建立NEC模型,对新生1日龄Wistar大鼠予100%二氧化碳,5min后再予以100%氧气,5min后放回母鼠身边喂养,第4天处死,取肠组织待检。新生鼠40只随机分为五组,每组8只,A组为NEC模型后予以腹腔注射ITF0.5mg;B组为NEC模型后予以皮下注射ITF0.2mg;C、D组为NEC模型后予以腹腔注射生理盐水分别为0.5ml和0.2ml;E组为正常对照组。取近回盲部1~2cm肠道组织固定包埋、切片、HE染色做病理学检查及免疫组化观察iNOS的表达,其他肠道组织制备组织匀浆取上清液检测TNF-α、NO的含量。结果A、B及E组iNOS呈弱阳性表达,C、D组iNOS呈强阳性表达;A、B组组织匀浆中TNF-α的含量各为[(30.515±2.731)和(32.229±4.978)pg/mg·pro]均较C、D组[(39.957±8.283)、(39.960±8.374)pg/mg·pro]明显下降(P<0.01),与E组[(33.523±6.752)pg/mg·pro]差异无统计学意义(P>0.05);A、B组组织匀浆中NO的含量各为[(0.37±0.07)和(0.54±0.08)μmol/mgtissue]较C、D组[(0.76±0.01)和(0.82±0.04)μmol/mgtissue]明显下降(P<0.01),与E组[(0.41±0.02)μmol/mgtissue]差异无统计学意义(P>0.05);A、B组间及C、D组间TNF-α、NO含量差异无统计学意义(P>0.05);病理切片显示C、D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;A、B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。结论通过腹腔和皮下注射ITF可以减轻NEC后的肠道炎症反应,ITF有可能为治疗NEC提供新的方法。
Objective To investigate the effect of intestinal trefoil factor (ITF) on iNOS, TNF-α and NO in intestinal mucosa of neonatal rats with necrotizing enterocolitis (NEC) and whether ITF has a protective effect on NEC . Methods The model of NEC was established. The newborn Wistar rats were given 100% carbon dioxide for 5 days and then 100% oxygen for 5 minutes. After 5 minutes, the rats were fed back to their mother rats and sacrificed on the 4th day. The intestine was taken for examination. Forty newborn rats were randomly divided into five groups with 8 rats in each group. Group A received intraperitoneal injection of ITF 0.5 mg after NEC model; Group B received ITF 0.2 mg subcutaneously after NEC model; Group C and D received NEC model Intraperitoneal injection of saline were 0.5ml and 0.2ml; E group was normal control group. Nearly ileocecal 1 ~ 2cm intestinal tissue fixed embedding, sliced, HE staining and immunohistochemistry to observe the expression of iNOS, other intestinal tissue tissue homogenate supernatant was taken to detect TNF-α, NO Content. Results The expression of iNOS in group A, B and E was weakly positive, while that in group C and D was strongly positive. The levels of TNF-α in group A and B were (30.515 ± 2.731) and (32.229 ± 4.978) pg / mg · pro] were significantly lower than those in group C [(39.957 ± 8.283), (39.960 ± 8.374) pg / mg · pro] (P> 0.05). The contents of NO in group A and B were (0.37 ± 0.07) and (0.54 ± 0.08) μmol / mgtissue respectively, compared with those in group C and D [ (0.41 ± 0.02) μmol / mgtissue] (P <0.05); Group A, B (0.76 ± 0.01) and (0.82 ± 0.04) μmol / There was no significant difference in the contents of TNF-α and NO between group C and group D (P> 0.05). Pathological examination showed that the damage of intestinal wall was observed in HE staining of C and D group. The score of the median score of 3 points; A, B group a small amount of intestinal epithelial shedding, the top of the villous necrosis, the pathological score of the median score of 1 point. Conclusions Intraperitoneal and subcutaneous injection of ITF can reduce intestinal inflammatory response after NEC, and ITF may provide a new method for the treatment of NEC.