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1 and clarify the differential roles of parent AN and its metabolites.Thus,this study would provide scientific evidence for the protection of workers and other population exposed to AN in the workplaces and general daily-life environment, especially for interaction between other medications and related chemicals. Method This study was divided into two parts.One is in vivo animal experiment;

the other is in vitro incubation assays.The subjects of animal experiments were SD rats, and dosages and routes of administration were selected based on our previous studies. The AN intraperitoneal(ip)injection were conducted at the dosages of

25 mg/kg,50 mg/kg and

75 mg/kg,and the rats were decapitated at 0.5 h,1 h,3 h,6 h,12 h and

24 h after AN injection.The control group was only ip injected with normal saline.The behavioral alterations were blindly recorded,and the level of hepatic CN'

,the activity of CcOx and CYP2E1 and the expression of CYP2E

1 protein were also determined. In addition,Potassium cyanide(KCN)selected as one of key metabolites of AN by CYP2E

1 was administrated to rats at dose of

3 mg/kg and Put to death at

1 5 min,0.5 h,1 h,3 h,and

6 h after KCN administration.Control group was only injected with normal saline.The in vitro experiments were divided into in vitro blank control group (in vitro spontaneous decline of CYP2E

1 activity),AN alone incubation group and AN metabolite incubation group(preincubated AN with NADPH).The alterations of III 丙烯腈急性染毒对大鼠肝脏细胞色素P450 2E1酶活性影响的研究 CYP2E

1 activity was measured over different times(0 rain,5 min,l

0 min,20 min,40 min,60 min)and doses(0Ⅳg,5 Irtg/g,1

0 I.tg/g,25 I-tg/g,50 I,tg/g,75 I.tg/g microsomal proteins). Results(1)All rats―treated with various dosages of AN showed,to some extent, over cholinergic toxicity symptoms:tearing,salivation,reddish ears and some rats had diarrhea,nose bleeding;

In the

75 mg/kg group,some of the rats expressed convulsions,purple lips,limbs weakness,shortness of breath,loss of righting reflex. (2)The results of CN.level in liver showed that,with increasing of exposure time, CN'

level is at first increased and subsequently decreased,and the higher dose of AN is,the higher the CN.level is.The hepatic contents of CN.remain constant from

1 5 min to 6h after KCN injection (3)The results of CcOx activity in liver showed that,with increasing exposure time, CcOx activity shows a trend first decreased and then recovered,and after 3h exposure, each group recovers to normal level. 江苏大学硕士学位论文 further study. Key words:acrylonitrile,CYP2E 1,acute poisoning,liver V 丙烯腈急性染毒对大鼠肝脏细胞色素P450 2E1酶活性影响的研究 VI 江苏大学硕士学位论文 目录

第一章绪论…1 1.1丙烯腈…1 1.1.1丙烯腈的接触机会及接触途径…l 1.1.2丙烯腈的代谢途径…1 1.1.3丙烯腈的毒性…2 1.2细胞色素P450 2El…3 1.2.1 CYP2El的研究史…3 1.2.2 CYP2El功能…4 1.2.3细胞色素P450活性的研究方法…4 1.2.4影响CYP2E1表达及活性的因素…4 1.3丙烯腈与细胞色素P450 2El…6

第二章研究目的意义、方法和实验设计方案…7 2.1研究目的及意义…7 2.2研究主要内容…7 2.3研究方法及主要观察指标…7 2.4实验设计方案…9

第三章急性丙烯腈染毒对大鼠肝脏CYP2EI酶活性的影响――体内实验……11 3.1主要试剂、仪器和实验方法…11 3.1.1主要试剂…1l 3.1.2主要仪器…12 3.1.3实验方法…12 3.2结果…17 3.2.1大鼠行为变化…17 3.2.2 AN急性染毒对大鼠肝组织中CYP2E1酶活性的影响……………….17 3.2.3 AN急性染毒对大鼠肝脏CYP2E1蛋白表达的影响………………….18 3.2.4 AN急性染毒对大鼠肝组织中CN.含量的影响…一20 3.2.5 AN急性染毒对大鼠肝组织中细胞色素C氧化酶活性的影响……….21 3.2.6氰化钾(KCN)染毒对对大鼠肝组织中CYP2E1酶活性的影响………22 3.3讨论…23