铁死亡在肝缺血-再灌注损伤大鼠肠损伤中的作用

Role of ferroptosis in intestinal injury induced by hepatic ischemia-reperfusion in rats

中文关键词: 铁死亡  缺血-再灌注  肠损伤

英文关键词: Ferroptosis  Ischemia reperfusion  Intestinal injury

中文摘要:

      
目的 探究铁死亡在肝缺血-再灌注损伤（HIR）大鼠肠损伤中的作用。
方法 选择健康清洁级雄性SD大鼠40只，周龄8～10周，体重220～260 g。采用随机数字表法将大鼠分成四组：假手术组（S组）、铁死亡抑制剂Ferrostatin-1+假手术组（SF组）、HIR模型组（IR组）和铁死亡抑制剂Ferrostatin-1+HIR模型组（IF组），每组10只。S组腹腔注射等容量0.02%DMSO，30 min后行假手术，仅进行开腹、分离第一肝门和关腹处理。SF组腹腔注射Ferrostatin-1（溶于0.02%DMSO）5 mg/kg，30 min后行假手术。IR组腹腔注射等容量0.02%DMSO，30 min后制备HIR模型。IF组腹腔注射Ferrostatin-1（溶于0.02%DMSO）5 mg/kg，30 min后制备HIR模型。于再灌注后8 h处死大鼠，取小肠组织，采用ELISA法检测肠组织丙二醛（MDA）、谷胱甘肽（GSH）和铁（Fe²⁺）浓度；采用速率法检测血清中ALT和AST活性；采用Western blot法检测谷胱甘肽过氧化物酶4（GPX4）、铁蛋白重链1（FTH1）和长链脂酰辅酶A合成酶4（ACSL4）蛋白含量；采用HE染色观察肝组织和小肠组织病理损伤并进行评分；采用透射电镜观察小肠组织线粒体形态。
结果 与S组比较，IR组和IF组MDA和Fe²⁺浓度、ACSL4蛋白含量、小肠组织病理损伤评分明显升高（P<0.05），GSH浓度、GPX4和FTH1蛋白含量明显降低（P<0.05），ALT和AST活性明显增强（P<0.05）。与IR组比较，IF组MDA和Fe²⁺浓度、ACSL4蛋白含量、小肠组织病理损伤评分明显降低（P<0.05），GSH浓度、GPX4和FTH1蛋白含量明显升高（P<0.05），ALT和AST活性明显减弱（P<0.05）。S组和SF组上述指标差异均无统计学意义。
结论 铁死亡参与肝缺血-再灌注大鼠肠氧化应激损伤过程，抑制铁死亡可以减轻肝缺血-再灌注损伤导致的肠损伤。

英文摘要:

      
Objective To investigate the role of ferroptosisin intestinal injury induced by hepatic ischemia-reperfusion in rats.
Methods Forty healthy SPF male adult SD rats, aged 8-10 weeks, weighing 220-260 g, were randomly divided into four groups: sham operation group (group S), Ferrostatin-1 pretreatment + sham operation group (group SF), hepatic ischemia-reperfusion (HIR) group (group IR), Ferrostatin-1 pretreatment + HIR group (group IF), 10 rats in each group. Group S injected 0.02% DMSO intraperitoneally 30 minutes before sham operation, group SF injected Ferrostatin-1 (dissolved in 0.02% DMSO) 5 mg/kg intraperitoneally 30 minutes before sham operation, group IR injected 0.02% DMSO intraperitoneally 30 minutes before establishing HIR model, group IF injected Ferrostatin-1 (dissolved in 0.02% DMSO) 5 mg/kg intraperitoneally 30 minutes before establishing HIR model. The rats were killed 8 hours after reperfusion. The concentrations of malondialdehyde (MDA), glutathione (GSH), and Fe²⁺ in intestinal tissue were detected by ELISA. The determination of serum ALT and AST activities were determined by velocity method. The contents of glutathione peroxidase 4 (GPX4), ferritin heavy chain 1 (FTH1), and long chain acyl coenzyme A synthase 4 (ACSL4) were detected by Western blot. HE staining was used to observe the pathological damage of liver and intestine, injury scores of intestine were scored. Transmission electron microscopy was used to observe the morphological changes of mitochondria in intestinal tissue.
Results Compared with group S, the concentrations of MDA and Fe²⁺, content of ACSL4 protein, intestinal tissue injury scores were significantly increased, the concentration of GSH,content of GPX4 and FTH1 protein were significantly decreased, ALT and AST activities were significantly enhanced in groups IR and IF (P < 0.05). Compared with group IR, the concentrations of MDA and Fe²⁺, content of ACSL4 protein, intestinal tissue injury scores were significantly decreased, the concentration of GSH,content of GPX4 and FTH1 protein were significantly increased, ALT and AST activities were significantly weakened in group IF (P < 0.05). There were no significant differences in all above indexes between groups S and SF.
Conclusion Ferroptosis is involved in intestinal oxidative stress injury during hepatic ischemia-reperfusion in rats. Inhibition of ferroptosis can reduce intestinal injury caused by hepatic ischemia-reperfusion injury.