马度米星铵对鲫鱼肝脏细胞色素P450酶系的影响  

Effects of maduramicin on hepatic cytochrome P450 enzymes of Carassius auratus

作　　者：杨丹 宋昕昊 季春雷 彭麟[1] 高修歌[1] 左儒楠 季辉[1] 江善祥[1] YANG Dan;SONG Xinhao;JI Chunlei;PENG Lin;GAO Xiuge;ZUO Runan;JI Hui;JIANG Shanxiang(College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China)

机构地区：[1]南京农业大学动物医学院,江苏南京210095

出　　处：《南京农业大学学报》2020年第5期927-934,共8页Journal of Nanjing Agricultural University

基　　金：国家自然科学基金项目(31672612)。

摘　　要：[目的]本文旨在研究鲫鱼肝脏细胞色素P450酶系对马度米星铵的响应。[方法]采用半静态染毒法,分别设置0.112、0.224和0.448 mg·L^-1马度米星铵处理组和助溶剂对照组,在染毒后7、14、21和28 d,测定肝脏中细胞色素P4501A(CYP1A)、3A(CYP3A)及4T(CYP4T)基因mRNA表达水平,同时测定7-乙氧基异吩唑酮-脱乙基酶(EROD)、氨基比林-N-脱甲基酶(APND)、苯胺-4-羟化酶(AH)、红霉素-N-脱甲基酶(ERND)、NADPH-细胞色素C还原酶(NCCR)的活性及细胞色素P450(CYP450)的含量;通过体外肝微粒体孵育法评价马度米星铵对CYP1A活性的影响。[结果]马度米星铵对CYP1A、CYP3A和CYP4T mRNA表达水平有一定诱导作用;0.448 mg·L^-1马度米星铵组CYP450含量在染毒后14 d显著低于对照组(P<0.05);0.224和0.448 mg·L^-1马度米星铵组APND和NCCR活性在染毒后14 d显著低于对照组(P<0.05);0.224和0.448 mg·L^-1马度米星铵组EROD活性在染毒后14和21 d显著低于对照组(P<0.05);试验期间,马度米星铵组ERND活性均低于对照组;各染毒组的AH活性呈先升高后降低的趋势;体外肝微粒体孵育体系中,马度米星铵对CYP1A活性的抑制作用小于25%。[结论]马度米星铵对鲫鱼肝微粒体CYP1A活性无影响;马度米星铵对ERND活性的抑制作用可能与CYP3A的转录水平无关;ERND对马度米星铵较为敏感,适合作为马度米星铵污染水体的生物标志物。[Objectives]The paper aimed to study the response of cytochrome P450 enzymes of Carassius auratus liver to maduramicin.[Methods]Semi-static poisoning method was used to expose Carassius auratus to 0.112,0.224,0.448 mg·L^-1 maduramicin for 28 days,and a cosolvent-control group was set up.On the 7,14,21 and 28 days after exposure,the liver of Carassius auratus was excised and homogenized,and the mRNA expression of cytochrome P4501A(CYP1A),cytochrome P4503A(CYP3A),and cytochrome P4504T(CYP4T)in Carassius auratus were determined by RT-qPCR.Furthermore,the activities of cytochrome P450(CYP450)enzymes such as 7-ethoxyresorufin-o-deethylase(EROD),aminopyrine-N-demethylase(APND),aniline hydroxylase(AH),erythromycin-N-demethylase(ERND)and NADPH cytochrome C reductase(NCCR),as well as the content of CYP450 in the liver were determined.The incubation system was established to investigate the effects of maduramicin on CYP1A in Carassius auratus liver microsomes in vitro.[Results]The mRNA expression levels of CYP1A,CYP3A and CYP4T were up-regulated by maduramicin to some degree.The content of CYP450 of 0.448 mg·L^-1group was significantly inhibited after 14 days exposure(P<0.05).In addition,the activities of APND and NCCR in the liver were significantly inhibited by 0.224 and 0.448 mg·L^-1maduramicin on day 14(P<0.05).On days 14 and 21,the activity of EROD in the liver was significantly inhibited by 0.224 and 0.448 mg·L^-1maduramicin(P<0.05).During the exposure period,the activity of ERND was lower in the experimental groups than in the control group.The activity of AH in the test group increased first and then decreased.In vitro liver microsomes incubation system,the inhibitory effect of maduramicin on CYP1A was less than 25%.[Conclusions]Maduramicin has no effect on CYP1A activity of Carassius auratus liver microsomes.The inhibition of ERND activity by maduramicin may not be regulated by the regulation of CYP3A transcription.In addition,ERND is sensitive to maduramicin and can be used as biomarker for maduramicin exposu

关 键 词：马度米星铵 鲫鱼 细胞色素P450 生化指标 肝微粒体 体外孵育 

分 类 号：S859.7[农业科学—临床兽医学]