基于TMT蛋白组学技术探讨氯化汞致小鼠肝脏损伤的机制  

Exploring the Mechanism of Mercury Chloride-induced Hepatotoxicity in Mice Based on TMT Proteomics

作　　者：曹鑫 张雅楠 毛侃敏 杨淼 郝丽萍[1] Cao Xin;Zhang Yanan;Mao Kanmin(Hubei Key Laboratory of Food Nutrition and Safety,Department of Nutrition and Food Hygiene,School of Public Health,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China)

机构地区：[1]华中科技大学同济医学院公共卫生学院营养与食品卫生学系,食品营养与安全湖北省重点实验室,武汉430030

出　　处：《华中科技大学学报（医学版）》2022年第5期585-591,共7页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：科技部国家重点研发计划“食品安全关键技术研发”专项(No.2018YFC1603101)。

摘　　要：目的利用蛋白组学技术筛选并鉴定氯化汞染毒后小鼠肝脏差异表达蛋白,为氯化汞的肝毒性作用机制研究提供线索。方法45只6周龄雄性C57BL/6小鼠随机分为5组:正常对照组,不同剂量氯化汞处理组,即低剂量组(1 mg/kg)、中低剂量组(4 mg/kg)、中高剂量组(8 mg/kg)、高剂量组(16 mg/kg)。氯化汞处理组每天灌胃给予不同剂量氯化汞,对照组给予等体积水,监测小鼠体重变化。第28天灌胃后处死小鼠,收集血清和肝脏组织,测定其血清丙氨酸氨基转移酶(alanine aminotransferase,ALT)、天门冬氨酸氨基转移酶(aspartate aminotransferase,AST)、肝脏谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)和过氧化氢酶(catalase,CAT)活性;苏木精-伊红(hematoxylin-eosin staining,HE)染色观察肝脏病理学变化;采用串联质谱标签(tandem mass tags,TMT)定量蛋白组学技术鉴定对照组与氯化汞高剂量组肝脏的差异表达蛋白(differentially expressed proteins,DEPs),并对其进行GO富集、KEGG通路富集分析。结果与对照组相比,中高剂量组与高剂量组小鼠体重降低(均P<0.05)、血清ALT活性显著增高、肝脏GSH-Px、CAT水平下降。HE染色结果显示氯化汞染毒可引起多种类型的肝细胞损伤。蛋白组学结果显示,与对照组相比,氯化汞高剂量组小鼠肝脏差异表达蛋白共有195个,其中上调99个、下调96个;GO富集分析显示氯化汞参与小鼠肝脏的糖代谢负调控、天冬氨酸家族氨基酸代谢等生物学过程;KEGG通路富集分析显示,氯化汞染毒可通过影响CYP450介导的外源化合物代谢、类固醇激素代谢、花生四烯酸代谢、胆汁分泌等信号通路产生肝脏毒性。结论氯化汞染毒严重影响小鼠肝脏多种蛋白的表达,导致相关代谢通路紊乱。Objective To investigate the mechanism of hepatotoxicity of mercury chloride on mice via proteomics.Methods Forty-five 6-week-old male C57BL/6 mice were randomly divided into 5 groups:control group,low-dose group(1 mg/kg),low-medium-dose group(4 mg/kg),medium-high-dose group(8 mg/kg),high-dose group(16 mg/kg).Mice in 4 treatment groups were given different doses of mercury chloride via oral gavage.The control group was given an equal amount of distilled water once a day.During the experiment,the body weight of the mice was observed and recorded dynamically.The mice were killed after the administration on the 28th day to collect serum and liver samples.Serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),liver glutathione peroxidase(GSH-Px),catalase(CAT)levels were tested.The pathological changes of the liver were observed through hematoxylin-eosin(HE)staining.Differentially expressed proteins(DEPs)between control and high-dose groups were identified by tandem mass tags(TMT)proteomics.GO and KEGG enrichment analyses were taken on DEPs.Results Compared with the control group,the body weight of mice decreased in the medium-high-dose group and high-dose group(both P<0.05).The serum ALT was significantly increased while the liver GSH-Px and CAT were significantly decreased in the medium-high-dose and high-dose groups(both P<0.05).HE staining result revealed that mercury chloride could cause several types of hepatocyte damage.A total of 195 DEPs were detected via proteomics,including 99 upregulated DEPs and 96 downregulated DEPs.GO enrichment analysis showed that mercury chloride was involved in the negative regulation of cellular carbohydrate metabolism,aspartate family amino acid metabolism,and other biological processes.KEGG pathway enrichment analysis showed that mercury chloride exposure could induce hepatotoxicity by affecting several signaling pathways,such as CYP450-mediated heterologous compound metabolism,steroid hormone biosynthesis,arachidonic acid metabolism,bile secretion,etc.Conclusion Mer

关 键 词：氯化汞 肝脏毒性 蛋白质组学 串联质谱标签定量蛋白组学技术 

分 类 号：R595.2[医药卫生—内科学]