【摘要】 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。更多还原

【Abstract】 Objective To study the effects of intestinal trefoil factor(ITF) on inducible nitric oxide synthase(iNOS),tumor necrosis factor-α(TNF-α) and NO density in neonatal rat model of necrotizing enterocolitis(NEC). Methods Forty neonatal rats were randomly divided into five groups: Group A, NEC+ ITF 0.5 mg; Group B, NEC+ ITF 0.2 mg; Group C, NEC+ N.S 0.5 ml; Group D, NEC+ N.S 0.2 ml; Group E, normal control. NEC model was established as following method: one-day old Wistar rats were put into 100%carbon dioxide for 5 min and then 100% oxygen for 5 min before returned to their mothers. This was done once daily for 3 days. On the 4th day, all rats were sacrificed and the intestinal tissue located at the boundary of ileum and cecum was obtained for histological examination by HE staining and iNOS, TNF-α and NO level by immunohistopathology. Results iNOS was found negative in group A, B and E, but positive in group C and D. The density of TNF-α were significantly decreased in group A[(30.515±2.731) pg/mg·pro] and B[(32.229± 4.978) pg/mg·pro] than that in group C[(39.957±8.283) pg/mg·pro] and D[(39.960±8.374) pg/mg·pro, P <0.01] and no difference with that in group E[(33.523±6.752) pg/mg·pro, P > 0.05)]. The density of NO were significantly decreased in group A[(0.37±0.07) μmol/mg tissue] and B[(0.54±0.08) μmol/mg tissue] than that in group C[(0.76±0.01) μmol/mg tissue] and D[(0.82± 0.04) μmol/mg tissue ( P <0.01)], and no difference with that in group E[(0.41±0.02) μmol/mg tissue( P >0.05)]. The TNF-α and NO expression showed no difference among group A and B, group C and D. The pathological lesions indicate that severe intestinal tissue necrosis in group C and D, and mild in group A and B. Conclusions Intestinal inflammation could be ameliorated after ITF injection hypodermically or intraperitoneally. ITF may provide a brand-new way for the management of NEC in neonatal rats.更多还原