长期高铜暴露通过影响线粒体自噬和细胞焦亡诱导大鼠肝组织损伤

doi:10.11843/j.issn.0366-6964.2022.12.034

摘要/Abstract

摘要： 旨在探索高铜对大鼠肝组织损伤的作用机制。试验选取120只SD大鼠随机分为对照组、高铜I组、高铜II组、高铜III组和高铜IV组，分别连续每天按体重灌胃0、20、40、80、160 mg·kg^-1铜63 d，采集肝组织。使用ICP-MS测定肝组织铜含量，病理切片观察肝组织病理变化，透射电镜观察线粒体形态和线粒体自噬小体，RT-qPCR和Western blot检测肝组织NLRP3、Caspase-1、GSDMD、IL-1β、IL-18、Pink1、Parkin、LCB3、p62的mRNA和蛋白表达水平。结果显示，随着灌胃铜水平的增加，蓄积在肝组织中的铜含量呈剂量依赖性增加，且长期高水平铜暴露会造成明显的肝组织结构破坏；随着铜暴露水平的增加，线粒体自噬和细胞焦亡水平呈先上升后下降趋势；与对照组相比，高铜Ⅱ组、Ⅲ组中PINK1和LC3II/LC3I的mRNA和蛋白表达水平显著上升(P＜0.05)，高铜Ⅱ组中p62的mRNA和蛋白表达水平显著降低(P＜0.05)，高铜Ⅲ组NLRP3、Caspase-1、GSDMD、IL-1β的mRNA和蛋白表达水平显著上升(P＜0.05)，高铜IV组表现为下调。高铜长期暴露可通过影响线粒体自噬和细胞焦亡诱导大鼠肝组织损伤。

关键词: 铜, 大鼠, 肝, 细胞焦亡, 线粒体自噬

Abstract: The mechanism of high levels of copper on liver injury in rats was investigated in this research. One hundred and twenty SD rats were randomly divided into 5 groups, including control group, high level copper group I, high level copper group Ⅱ, high level copper group Ⅲ and high level copper group Ⅳ. The liver tissues were collected from the rats which were continuously intragastric administration of 0, 20, 40, 80 and 160 mg·kg^-1 copper for 63 days, respectively. The copper content in liver was determined by ICP-MS, and the pathological changes of liver were observed by pathological sections. The mitochondrial morphology and mitophagosomes were observed by transmission electron microscopy (TEM). The mRNA and protein expression levels of NLRP3, Caspase-1, GSDMD, IL-1β, IL-18, Pink1, Parkin, LCB3 and p62 in liver were detected by RT-qPCR and Western blot, respectively. The results showed that the Cu content in liver exhibited a significant dose-dependent effect with increased copper concentration, and long-term exposure to high levels of copper would cause significant structural damage and pathological changes in liver. With the increase of copper exposure level, the levels of mitophagy and pyroptosis decreased after initially increasing. Compared with the control group, the mRNA and protein expression levels of mitophagy-related genes and proteins PINK1 and LC3II/LC3I in high level copper group II and high level copper group III increased significantly (P<0.05). The mRNA and protein expressions of p62 in high level copper group Ⅱ were significantly decreased (P<0.05), and the mRNA and protein expressions of NLRP3, Caspase-1, GSDMD and IL-1β were significantly increased in high level copper group III (P<0.05), but down regulated in high level copper group IV. High levels of copper exposure could induce liver injury in rats by affecting mitophagy and pyroptosis.

Key words: copper, rat, liver, pyroptosis, mitophagy

中图分类号:

S856.9

梁文清, 刘忠华, 常晓月, 何婷, 陈嘉, 马晓莉, 唐兆新, 余文兰. 长期高铜暴露通过影响线粒体自噬和细胞焦亡诱导大鼠肝组织损伤[J]. 畜牧兽医学报, 2022, 53(12): 4490-4500.

LIANG Wenqing, LIU Zhonghua, CHANG Xiaoyue, HE Ting, CHEN Jia, MA Xiaoli, TANG Zhaoxin, YU Wenlan. Long-term Exposure to High Levels of Copper Induced Liver Injury in Rats by Affecting Mitophagy and Pyroptosis[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(12): 4490-4500.

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