去铁胺对新生大鼠坏死性结肠炎的保护作用

Protective effect of deferoxamine on necrotizing colitis in neonatal rats

目的探讨去铁胺对新生大鼠坏死性结肠炎保护作用及其机制。方法30只SD新生大鼠按照随机数字表法分为对照组、模型组和去铁胺组,每组10只,模型组和去铁胺组大鼠采用常规配方乳灌胃建立坏死性结肠炎模型,对照组新生大鼠给予鼠乳。去铁胺组新生大鼠灌胃给予30 mg/(kg·d)去铁胺,对照组和模型组新生大鼠给予等体积生理盐水。治疗4 d后,观察3组大鼠的一般情况;采用苏木精-伊红(HE)染色分析3组大鼠的肠道病理学变化;采用酶联免疫吸附实验分析3组大鼠肠道组织炎性因子肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-12和IL-18表达水平;采放免法分析3组大鼠肠道组织氧化应激指标水平;采用蛋白质免疫印迹分析肠道组织谷胱甘肽过氧化物酶4(GPX4)和胱氨酸/谷氨酸反向转运体(SLC7A11)表达水平。荧光定量聚合酶链反应(PCR)分析肠道组织CXC型趋化因子配体1(CXCL1)和CXC趋化因子受体2(CXCR2)mRNA表达水平。组间比较采用单因素方差分析。结果去铁胺组大鼠体重变化[(1.11±0.11)g]明显高于模型组新生大鼠[(0.55±0.11)g],差异有统计学意义(t=11.220,P<0.05)。去铁胺组新生大鼠疾病活动指数评分(1.80±0.79)明显低于模型组大鼠(3.70±1.06),差异有统计学意义(t=4.549,P<0.05)。去铁胺组新生大鼠肠道组织TNF-α、IL-12和IL-18炎性因子水平[(45.63±11.32)、(40.03±5.97)、(67.62±7.27)pg/ml]明显低于模型组[(84.60±12.20)、(73.89±13.48)、(107.59±12.22)pg/ml],差异有统计学意义(t=7.025、6.890、8.437,P<0.05)。去铁胺组新生大鼠肠道组织CXCL1和CXCR2 mRNA表达水平(1.36±0.12、1.35±0.09)明显低于模型组(1.80±0.14、1.85±0.08),差异有统计学意义(t=6.968、11.970,P<0.05)。去铁胺组新生大鼠肠道组织SOD和GSH-PX活性水平[(45.65±9.27)、(47.53±6.78)U/mg]明显高于模型组[(26.86±4.51)、(22.74±3.79)U/mg],差异有统计学意义(t=5.468、9.571,P<0.05)。去铁胺组新生大鼠肠道组织GPX4和SLC7A11蛋白表达水平(0.89±0.04、0.64±0.06)明显高于模型组(0.51±0.19、0.38±0.11),差异有统计学意义(t=5.571、6.174,P<0.05)。结论去铁胺可显著抑制铁死亡,降低肠道组织炎性反应,进而保护结肠组织。

Objective To explore the protective effect and molecular mechanism of deferoxamine on necrotizing colitis in neonatal rats.MethodsA total of 30 SD neonatal rats were randomly divided into a control group,a model group,and a deferoxamine group using a random number table method,with 10 rats in each group.The rats in the model group and deferoxamine group were given conventional formula milk by gavage to establish a necrotizing colitis model,while neonatal rats in the control group were given rat milk.The neonatal rats in the deferoxamine group were given 30 mg/(kg·d)deferoxamine by gavage,while the control group and model group were given equal volume of physiological saline.After 4 days of treatment,the general conditions of rats in the three groups were observed.The intestinal pathological changes of rats in the three groups were analyzed using hematoxylin eosin(HE)staining.The expression levels of inflammatory factors[tumor necrosis factor-α(TNF-α),interleukin(IL)-12 and IL-18]in intestinal tissue of three groups were tested using enzyme-linked immunosorbent assay.The levels of oxidative stress indicators in the intestinal tissues of three groups were analyzed using immunoassay.The expression levels of glutathione peroxidase 4(GPX4)and cysteine/glutamate reverse transporter(SLC7A11)in intestinal tissue were analyzed by Western blotting.The CXC tchemokine ligand 1(CXCL1)and CXC chemokine receptor 2(CXCR2)mRNA expression levels in intestinal tissue were analyzed by fluorescence quantitative polymerase chain reaction(PCR).The comparison of inter group econometric data was conducted using one-way analysis of variance.ResultsThe weight change of the rats in the deferoxamine group[(1.11±0.11)g]was significantly greater than that of the newborn rats in the model group[(0.55±0.11)g,t=11.220,P<0.05].The disease activity index score of neonatal rats in the deferoxamine group(1.80±0.79)was significantly lower than that in the model group(3.70±1.06,t=4.549,P<0.05).The levels of inflammatory factors(TNF-α,IL-12 and IL-18)in the intestinal tissue of neonatal rats treated with deferoxamine[(45.63±11.32),(40.03±5.97),(67.62±7.27)pg/ml]were significantly lower than those in the model group[(84.60±12.20),(73.89±13.48),(107.59±12.22)pg/ml,t=7.025,6.890,8.437,P<0.05].The mRNA expression levels of CXCL1 and CXCR2 in the intestinal tissue of neonatal rats in the deferoxamine group(1.36±0.12,1.35±0.09)were significantly lower than those in the model group(1.80±0.14,1.85±0.08,t=6.968,11.970,P<0.05).The levels of SOD and GSH-PX activity in the intestinal tissue of neonatal rats in the deferoxamine group[(45.65±9.27),(47.53±6.78)U/mg]were significantly higher than those in the model group[(26.86±4.51),(22.74±3.79)U/mg,t=5.468,9.571,P<0.05].The expression levels of GPX4 and SLC7A11 proteins in the intestinal tissue of neonatal rats in the deferoxamine group(0.89±0.04,0.64±0.06)were significantly higher than those in the model group(0.51±0.19,0.38±0.11)(t=5.571,6.174,P<0.05).ConclusionDeferoxamine can significantly inhibit ferroptosis,reduce intestinal inflammation,and thus protect colon tissue.