【摘要】 为探讨丙烯腈（acrylonitrile,ACN）诱导的氧化应激对大鼠脾核转录因子NF-κB（nuclear transcription factor-κB,NF-κB）信号通路的影响,将60只无特定病原体（specific pathogen free,SPF）级健康成年雄性大鼠随机分为5组:对照组（玉米油）,低、中、高剂量染毒组(11.5、23、46 mg·kg^-1ACN)和N-乙酰-L-半胱氨酸（N-acetylcysteine,NAC）干预组(46 mg·kg^-1ACN+300mg·kg^-1NAC),1次/天,6天/周,连续灌胃4周。染毒结束后,次日处死大鼠,分离大鼠脾并称重。以分光光度法检测脾组织超氧化物歧化酶（superoxide dismutase,SOD）、谷胱甘肽过氧化物酶（glutathione peroxidase,GSH-Px）、谷胱甘肽（glutathione,GSH）、丙二醛（malondialdehyde,MDA）、过氧化氢酶（catalase,CAT）和总抗氧化能力（total antioxidant capacity,T-AOC）含量或活力;以Elisa法检测大鼠脾白介素6（IL-6）、白介素1β（IL-1β）、肿瘤坏死因子-α（TNF-α）水平;以Western Blot法检测脾NF-κB、IκB、p-IκB蛋白表达水平;以RT-PCR法检测NF-κB、IκB mRNA表达水平。结果显示,与对照组比较,ACN低、中剂量组大鼠脾湿重明显降低（P<0.05）; ACN中剂量组脾脏器系数降低（P<0.05）。中、高剂量组大鼠脾组织MDA含量显著增加（P<0.05）;高剂量组大鼠SOD、GSH-Px活力显著增加（P<0.05）;低剂量组大鼠CAT活力降低,高剂量组大鼠CAT活力增加（P<0.05）;低剂量组大鼠GSH含量、T-AOC活力均降低（P<0.05）。与高剂量组相比较,NAC组大鼠MDA、SOD、CAT、GSH、GSH-Px、T-AOC均降低（P<0.05）。与对照组比较,ACN中、高剂量组大鼠脾组织内IL-1β含量降低（P<0.05）,低、中、高剂量组大鼠TNF-α含量增加（P<0.05）。与高剂量组比较,NAC组TNF-α含量降低（P<0.05）。Western Blot结果显示,中、高剂量染毒组大鼠脾NF-κB、IκB、p-IκB蛋白表达水平与对照组比较均升高（P<0.05）。NAC组大鼠脾IκB、p-IκB蛋白表达水平与高ACN组比较降低（P<0.05）。RT-PCR结果显示,ACN各剂量染毒组大鼠脾NF-κB mRNA表达水平与对照组比较均升高（P<0.05）;中、高剂量染毒组大鼠脾IκB mRNA表达水平与对照组比较均升高（P<0.05）。NAC组大鼠脾NF-κB mRNA表达水平低于高剂量组（P<0.05）。结果表明ACN亚急性染毒可对大鼠脾产生氧化损伤和免疫系统响应,NAC可通过拮抗作用减轻损伤的程度。更多还原

【Abstract】 To investigate the effect of acrylonitrile（ ACN）-induced oxidative stress on nuclear transcription factor-κB（ NF-κB） signaling pathway in rats spleen,60 specific pathogen free（ SPF） grade healthy adult male rats were randomly divided into 5 groups: control group（ corn oil）,low-,medium-,and high-dose groups( 11.5,23,46 mg·kg^-1 ACN) and NAC groups( 46 mg·kg^-1 ACN + 300 mg·kg^-1 NAC),and were treated once/day,6 days/week,by intragastric gavage for 4 weeks. The spleen was isolated and weighed. The activities of superoxide dismutase（ SOD）,glutathione peroxidase（ GSH-Px）,and catalase（ CAT）,the contents of glutathione（ GSH）,malondialdehyde（ MDA） and total antioxidant capacity（ T-AOC） in spleen were detected by spectrophotometry. The interleukin 6（ IL-6）,interleukin 1β（ IL-1β）,tumor necrosis factor-α（ TNF-α） levels were determined by Elisa.The expressions of NF-κB,IκB and p-IκB were measured using western blotting and the expressions of NF-κB and IκB mRNA were determined by RT-qPCR. The results showed that the splenic wet weight of the rats were significantly decreased in the low-and middle-dose groups compared with the control group（ P<0.05）. The splenic coefficient was also significantly decreased in the middle-dose group（ P<0.05）. The contents of MDA in the spleen of rats were significantly increased in the medium-and high-dose groups（ P < 0. 05） and the activities of SOD and GSH-Px were significantly increased in the high-dose group（ P<0.05）. The activity of CAT and the contents of TAOC and GSH were all decreased in the low-dose group,whereas,the CAT activity was increased in the high-dose group（ P<0.05）.Meanwhile,the activities of SOD,CAT and GSH-Px and the contents of MDA,GSH and T-AOC were significantly decreased in the NAC group compared with the high-dose group（ P<0.05）. Compared with the control group,the contents of IL-1β were decreased in the middle-and high-dose groups,and the levels of TNF-αwere increased significantly in all dose groups（ P<0.05）. Compared with the high-dose group,the content of TNF-α was significantly decreased in the NAC group（ P<0.05）. Western Blot results showed that the expressions of NF-κB,IκB,and p-IκB were significantly increased in the medium-and high-dose groups compared with the control group（ P<0.05）. The expressions of IκB and p-IκB in NAC group was decreased in the high ACN group（ P <0.05）. RT-PCR results showed that the expression of NF-κB mRNA were increased in the low-,medium-and highdose ACN groups compared with the control group（ P<0.05）. And the expressions of IκB mRNA were increased in the medium-and high-dose ACN groups compared with the control group（ P<0.05）. The expression of NF-κB mRNA was decreased in the NAC group compared with the high-dose group（ P<0.05）. Therefore,the present study suggests that subacute exposure to ACN could produce oxidative damage and immune system response in rats ’spleen,and NAC might reduce the degree of injury through antagonism.更多还原