铁死亡参与镉暴露鸡肝损伤的研究

doi:10.11843/j.issn.0366-6964.2023.02.035

摘要/Abstract

摘要： 旨在研究铁死亡在镉致鸡肝组织损伤中的作用。选取30只1日龄海兰白蛋公鸡，预饲7 d后，随机平均分为对照组（C组）和镉暴露试验组（Cd组），C组鸡饲喂基础日粮，Cd组鸡饲喂混有140 mg·kg^-1 CdCl₂的基础日粮。镉暴露后20、40和60 d取样，观察肝组织病理及超微病理学变化，检测肝功能酶活性，肝组织氧化应激水平，肝中炎症因子、铁死亡相关因子及线粒体融合相关因子表达水平；建立肝癌细胞系（LMH）染镉细胞模型，同时设立铁死亡激活剂和铁死亡抑制剂染镉细胞组，镉暴露24 h，检测细胞活性、炎症因子表达、铁死亡相关因子和线粒体融合相关因子表达。结果发现，Cd组鸡血清中肝功能酶活性明显升高；肝细胞排列紊乱、空泡变性甚至坏死，肝组织淤血，浆液-纤维素性渗出，炎性细胞浸润；肝细胞线粒体体积缩小、嵴断裂。肝中炎症因子LOX、NF-κB、TNF-α、IL-6和IL-1β mRNA相对表达上调，TNF-α、IL-6和IL-1β含量显著上升；过氧化物MDA含量明显升高、抗氧化酶GSH-Px与SOD活性显著降低；铁死亡代谢相关因子铁蛋白重链1（FTH1）和转铁蛋白受体（TFR） mRNA相对表达显著升高，铁死亡标志物谷胱甘肽过氧化物酶4（GPX4）表达极显著降低，长链脂酰辅酶A合成酶4（ACSL4）和环加氧酶2（PTGS2）表达极显著升高；线粒体融合相关因子视神经萎缩蛋白1（OPA1）、线粒体融合蛋白1/2（Mfn1/2） mRNA表达极显著降低。镉暴露及铁死亡激活剂均可引发LMH细胞铁死亡，促进炎性因子释放和线粒体功能障碍，导致细胞损伤；铁死亡抑制剂极显著地抑制了镉诱导的LMH细胞炎症因子释放和线粒体损伤，减轻镉导致的细胞活力降低。结果表明，镉暴露导致肝组织氧化应激，引起肝内脂质过氧化物蓄积、铁代谢紊乱，进而诱发铁死亡，促进肝炎性损伤。

关键词: 镉, 铁死亡, 肝损伤, 鸡

Abstract: The aim of this experiment was to study the effect of ferroptosis on cadmium-induced liver damage of chicken. After 7 d pre-feeding, thirty 1-day-old Hylan white chickens were randomly divided into control group (group C) and cadmium exposure test group (group Cd), with 15 chickens in each group. Chickens in group C were fed the basal diet, and chickens in group Cd were fed the diet supplemented with 140 mg·kg^-1 CdCl₂. Samples were taken at 20, 40 and 60 d after cadmium exposure to observe liver histopathological and ultra-pathological changes, and to detect liver functional enzyme activities, levels of inflammatory factors, oxidative stress, ferroptosis-related factors and mitochondrial fusion-related factors in the liver. Based on the establishment of liver cancer cell line (LMH) exposed to cadmium, groups of iron death activator and iron death inhibitor were put up as well, and the cell activity, expression of inflammatory factors, iron death related factors and mitochondrial fusion related factors were detected after 24 hours. The results showed that the activity of liver functional enzymes in the serum of chickens in the group Cd was significantly increased; Liver cells were disorder, vacuolar degeneration or even necrosis; Liver congestion, serousand fibrinous exudation, and inflammatory cell infiltration; The relative expression levels of inflammatory factors LOX, NF-κB, TNF-α, IL-6 and IL-1β mRNA in the liver were up-regulated, and the contents of TNF-α, IL-6 and IL-1β were increased; The content of peroxide MDA was significantly increased, the activities of antioxidant enzymes GSH-Px and SOD were significantly decreased; The relative expression levels of ferroptosis metabolism-related factors FTH1 and TFR mRNA were significantly increased levels. The expression levels of GPX4 was significantly decreased, and the expression levels of ACSL4 and PTGS2 were extremely significantly increased; Mitochondrial fusion-related factors OPA1, Mfn1/2 mRNA expression levels were significantly decreased. Cadmium exposure and ferroptosis activators can both induce ferroptosis in LMH cells, promote the release of inflammatory factors, cause mitochondrial dysfunction, and lead to cell damage; Ferroptosis inhibitor significantly inhibited the cadmium-induced release of inflammatory factors and mitochondrial damage in LMH cells, and alleviated the decrease in cell viability caused by cadmium. The results indicated that cadmium exposure caused the accumulation of lipid peroxides, the disturbance of iron metabolism, and ferroptosis, which eventually promoted inflammatory damage to the liver.

Key words: cadmium, ferroptosis, liver damage, chickens

中图分类号:

S858.316.9

陈敬宜, 于淼, 张金洋, 樊堃, 杨桂君, 葛铭, 张瑞莉. 铁死亡参与镉暴露鸡肝损伤的研究[J]. 畜牧兽医学报, 2023, 54(2): 787-802.

CHEN Jingyi, YU Miao, ZHANG Jinyang, FAN Kun, YANG Guijun, GE Ming, ZHANG Ruili. Study on the Involvement of Ferroptosis in Liver Injury of Cadmium-exposed Chickens[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(2): 787-802.

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