A diet high in red meat promotes ulcerative colitis in mice through ferroptosis pathway
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维普
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目的 探讨铁死亡在高红肉饮食促进溃疡性结肠炎(Ulcerative colitis,UC)发生中的作用机制.方法 36只雄性Balb/c小鼠中随机选取18只,给予标准饲料,另外18只小鼠给予含60％红肉饲料.8周后,给予标准饲料的18只小鼠分为对照组、UC模型组和UC模型+铁死亡抑制剂(Fer-1)组,每组各6只;给予含60％红肉饲料的18只小鼠分为高红肉组、高红肉+UC模型组和高红肉+UC模型+Fer-1组,每组各6只.除对照组、高红肉组外,其余组予小鼠3％葡聚糖硫酸钠(Dextran sodium sulfate,DSS)构造UC小鼠模型,UC模型+Fer-1组和高红肉+UC模型+Fer-1组造模同时给予铁死亡抑制剂(Ferrostatin-1,Fer-1)腹腔注射,连续9 d.比较UC小鼠疾病活动指数(DAI)评分和HE染色评价小鼠UC严重程度,Elisa、RT-qPCR和透射电镜检测小鼠亚铁离子(Ferrous ion,Fe2+)、活性氧(Reactive oxygen species,ROS)、丙二醛(Malondialdehyde,MDA)、谷胱甘肽(Glutathione,GSH)、谷胱甘肽过氧化物酶 4(Glutathione peroxidase 4,GPX4)、铁蛋白重链1(Ferritin heavy chain1,FTH1)和酰基辅酶A合成酶长链家族成员4(Acyl-CoA syn-thetase long-chain family member 4,ACSL4)的铁死亡以及肿瘤坏死因子 α(Tumor necrosis fac-tor-α,TNF-α)和白介素6(Interleukin 6,IL-6)等炎症相关因子的表达,并观察小鼠结肠肠上皮细胞线粒体形态的变化.结果 与UC模型组相比,高红肉+UC模型组小鼠DAI评分显著升高(P＜0.05),结肠黏膜结构被完全破坏,结肠肠上皮细胞线粒体空泡化严重且线粒体膜破裂、嵴消失;小鼠结肠中Claudin1、Occludin和ZO-1mRNA表达水平明显降低(P＜0.05);在小鼠血清中Fe2+、ROS、MDA、TNF-α及IL-6表达水平明显升高,GSH表达水平明显降低(P＜0.05);在小鼠结肠组织中GPX4和FTH1 mRNA的表达水平明显降低,ACSL4 mRNA的表达水平明显升高(P＜0.05).Fer-1干预后,结肠炎小鼠铁死亡及炎症相关指标降低,抗氧化指标上升(P＜0.05).结论 高红肉饮食可加重UC模型小鼠肠道炎症,其机制可能是通过调控铁死亡通路,增加结肠炎症的表达,促进UC的发生发展,Fer-1对高红肉饮食诱导的铁死亡和炎症有显著的改善作用.
Objective To investigate the mechanism of ferroptosis in the promotion of ulcerative colitis(UC)by a high red meat diet.Methods 18 Balb/c male mice were randomly selected from 36 mice and given a standard diet,while the other 18 mice were given 60％red meat diet.After 8 weeks,18 mice given a standard diet were divided into control group,UC model group,and UC model+Fer-1 group,with 6 mice in each group;18 mice fed with 60％red meat were divided into high-red meat group,high red meat+UC model group,and high red meat+UC model+Fer-1 group,with 6 mice in each group.Except for the control group and the high red meat group,the remaining groups were given 3％Dextran Sodium Sul-fate(DSS)to construct UC mouse models,and UC model+Fer-1 group and high red meat+UC model+Fer-1 group were given intraperitoneal injections of Ferrostatin-1(Fer-1)for 9 consecutive days at the same time as modeling.Comparison of disease activity index(DAI)scores and HE staining in UC mice were used to evaluate the severity of UC in mice,and Elisa,RT-qPCR and transmission electron microsco-py were used to detect ferrous ion(Fe2+),reactive oxygen species(ROS),malondialdehyde(MDA),glu-tathione(GSH),glutathione peroxidase 4(GPX4),ferritin heavy chain1(FTH1)and Acyl-CoA syn-thetase long-chain family member 4(ACSL4)for ferroptosis as well as the expression of inflammation-re-lated factors such as tumor necrosis factor-α(TNF-α)and interleukin 6(IL-6)and observed the changes in mitochondrial morphology of mouse colon intestinal epithelial cells.Results Compared with UC model group,the DAI scores of mice in the high red meat+UC model group were significantly higher(P＜0.05),the structure of colonic mucosa was completely destroyed,and mitochondrial vacuolization of co-lonic intestinal epithelial cells were severe with rupture of the mitochondrial membranes and disappearance of the cristae.The expression levels of Claudin 1,Occludin and ZO-1 mRNA in the mouse colon were sig-nificantly lower(P＜0.05).The expression levels of Fe2+,ROS,MDA,TNF-α and IL-6 were signifi-cantly elevated in the sera of mice,and the expression of GSH was significantly decreased(P＜0.05).The expression levels of GPX4 and FTH1 mRNA were significantly reduced in mice colon tissues,and the expression of ACSL4 mRNA was significantly elevated(P＜0.05).Ferroptosis and inflammation-related indexes were reduced,antioxidant indexes were increased in mice with colitis after Fer-1 intervention(P＜0.05).Conclusion High red meat diet exacerbates intestinal inflammation in UC model mice by a mecha-nism that may promote the development of UC by modulating the ferroptosis pathway and increasing the expression of colonic inflammation,and Fer-1 has a significant ameliorating effect on ferroptosis and in-flammation induced by high red meat diet.
维普
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