苯并[b]荧蒽亚急性经口染毒的大鼠肝脏毒性以及转录组改变  

Hepatotoxicity and transcriptome changes in rats with subacute oral exposure to benzo [b]fluoranthene

作　　者：赵晨阳 吴倩 张云波[1] 李刚[1] 那晓琳[1] ZHAO Chen-yang;WU Qian;ZHANG Yun-bo;LI Gang;NA Xiao-lin(Public Health College,Harbin Medical University,Harbin 150086,China)

机构地区：[1]哈尔滨医科大学公共卫生学院,哈尔滨150086

出　　处：《环境卫生学杂志》2022年第11期777-784,789,共9页JOURNAL OF ENVIRONMENTAL HYGIENE

摘　　要：目的分析小剂量苯并[b]荧蒽(benzo[b]fluoranthene,BbF)经口暴露引起的大鼠肝脏转录组变化,探讨BbF的肝脏毒性及可能机制。方法选取16只体重在90~100 g的雄性SD大鼠,适应性培养后根据体重将大鼠随机分为实验组与对照组,两组分别以溶剂、1μg/kg BbF的剂量灌胃,每周两次,连续4周。暴露结束后处死取血与肝脏组织,分别测定血清指标和氧化应激水平;随后利用转录组测序(RNA-Sequencing,RNA-Seq)分析大鼠肝脏基因表达变化,GO富集分析差异基因的主要功能,蛋白互作(protein-protein interaction,PPI)网络分析和Clue GO富集分析可视化差异基因的相互作用,实时定量聚合链式反应(quantitative real-time PCR,RT-PCR)验证转录组结果。结果暴露结束后对照组与实验组动物脏器系数分别为2.60%±0.23%和3.12%±0.25%。与对照组相比,实验组大鼠肝脏系数显著增大(P<0.01),体重差异无统计学意义(P>0.05)。血清学指标显示,两组动物血清谷丙转氨酶(alanine transaminase,ALT)与谷草转氨酶(aspartate aminotrans ferase,AST)的活性差异无统计学意义(P>0.05)。实验组动物肝脏过氧化氢(hydrogen peroxide,H 2O 2)含量为(34.46±2.04)mmol/g,过氧化氢酶(catalase,CAT)活性为(14.03±2.38)U/mg,超氧化物歧化酶(superoxide dismutase,SOD)为(40.70±3.74)U/mg;与对照组相比,实验组动物H 2O 2含量增加(P<0.05),CAT与SOD活性显著降低(P<0.01),而丙二醛(malondialdehyde,MDA)含量差异无统计学意义(P>0.05)。两组动物间转录组测序结果显示存在394个差异基因,其中229个上调,165个下调。GO富集分析显示氧化还原过程富集到差异基因最多,并且多个蛋白折叠相关条目被显著富集。PPI网络分析和Clue GO富集分析发现,差异基因的关键节点为Hspa5,Acox2和Ehhadh,关键基因网络主要涉及细胞未折叠蛋白的反应和抗氧化能力。结论低BbF暴露引起大鼠肝脏轻微氧化应激,干扰内质网功能;机体保护性触发未折�Objective To investigate liver transcriptome changes in rats with oral exposure to low-dose benzo[b]fluoranthene(BbF), and to explore the hepatotoxicity of BbF and its possible mechanism. Methods A total of 16 male SD rats, with a body weight of 90-100 g, were selected, and were randomly divided into experimental group and control group based on body weight after adaptive feeding. The two groups were treated with solvent and 1 μg/kg BbF, respectively, by gavage, twice a week for 4 consecutive weeks. The rats were sacrificed at the end of exposure to collect blood and liver tissue samples for determination of serological parameters and oxidative stress levels. RNA sequencing was used to investigate changes of gene expression in livers, gene ontology(GO) enrichment analysis was used to investigate the main functions of differentially expressed genes, and the protein-protein interaction(PPI) network and Clue GO analysis were used to visualize the interaction between differentially expressed genes;quantitative real-time PCR(RT-PCR) was used to validate the results of transcriptome. Results At the end of the exposure, the organ coefficient was 2.60%±0.23% in the control group and 3.12%±0.25% in the experimental group. Compared with the control group, the experimental group had a significantly higher organ coefficient of the liver(P<0.01), while there was no significant difference in body weight between the two groups(P>0.05). The analysis of serological parameters showed that there were no significant differences in the contents of alanine transaminase(ALT) and aspartate aminotransferase(AST) between the two groups(P>0.05). In the experimental group, the content of hydrogen peroxide(H2O2) was 34.46±2.04 mmol/g, the activity of catalase(CAT) was 14.03±2.38 U/mg, and the activity of superoxide dismutase(SOD) was 40.70±3.74 U/mg. Compared with the control group, the experimental group had a significant increase in the content of H2O2(P<0.05) and significant reductions in activities of CAT and SOD(P<0.01), and there

关 键 词：多环芳烃 苯并[b]荧蒽(BbF) 转录组学 肝脏毒性 未折叠蛋白反应 

分 类 号：R122[医药卫生—环境卫生学]