摘要:
为探索丙烯腈(acrylonitrile, ACN)诱导的氧化应激对大鼠睾丸NF-κB信号通路的影响,将50只SPF级健康SD雄性大鼠按体重随机分为12.5、25、50 mg·kg-1 ACN染毒组,50 mg·kg-1 ACN+300 mg·kg-1 N-乙酰半胱氨酸(N-acetylcysteine, NAC)干预组(NAC干预组),对照组(给予等体积玉米油),每组10只,灌胃,1次/天,6天/周,共90 d。可见光分光光度法检测睾丸组织中超氧化物歧化酶(superoxide dismutase, SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase, GSH-Px)、还原型谷胱甘肽/氧化型谷胱甘肽比值(glutathione/oxidized glutathione, GSH/GSSG)、丙二醛(malondialdehyde, MDA)。免疫荧光染色法检测睾丸核因子-κB(nuclear factor κB, NF-κB)激活及核转移。Western Blot检测睾丸p65、IκB蛋白表达。结果显示,低、高剂量染毒组大鼠睾丸GSH/GSSG比值、GSH-Px酶活性与对照组比较降低(P<0.05)。中、高剂量染毒组大鼠睾丸MDA含量与对照组比较升高(P<0.05)。NAC干预组大鼠睾丸MDA含量与高ACN组比较降低(P<0.05);NAC干预组大鼠睾丸GSH/GSSG比值与高ACN组比较升高(P<0.05);免疫荧光结果显示,高ACN组大鼠睾丸NF-κB被激活,并转移入细胞核。NAC干预组与高ACN组比较p65蛋白表达及核转移显著减少。Western Blot结果显示,高剂量染毒组大鼠睾丸p65蛋白表达与对照组比较升高(P<0.05),IκB蛋白表达与对照组比较降低(P<0.05);NAC干预组大鼠睾丸p65蛋白表达与高ACN组比较降低,IκB蛋白表达与高ACN组比较升高,差异有统计学意义(P<0.05)。结果表明丙烯腈引起的氧化应激激活了大鼠睾丸生殖细胞NF-κB信号通路。
关键词:
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丙烯腈
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大鼠
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睾丸
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氧化应激
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NF-κB
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p65
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IκB
Abstract:
In order to study the effects of acrylonitrile (ACN)-induced oxidative stress via NF-κB signaling pathway in rat testis, fifty Sprague-Dawley male rats in SPF degree were randomly divided into 5 groups according to the body weight, and were treated with 0, 12.5, 25, 50 mg·kg-1 ACN or 50 mg·kg-1 ACN + 300 mg·kg-1 N-acetylcysteine (NAC) (NAC intervention group) respectively, via gavage, 6 days per week for 90 days. The activity of testicular superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), glutathione (GSH) and contents of maleic dialdehyde (MDA) were determined by visible spectrophotometry. The activation and nuclear translocation of testicular nuclear factor κB (p65) was detected by immunofluorescence. The testicular protein level of p65 and IκB were detected by Western Blot. The results showed that in comparison with control grpup, the ratio of GSH/GSSG and the activity of GSH-Px were significant decreased in low and high concentration of ACN group. The contents of MDA were significant increased in medium and high concentration of ACN groups. In NAC intervention group, the contents of MDA were significant decreased, while the ratio of GSH/GSSG was significantly increased as compared with high concentration of ACN group. Results of immunofluorescence showed that NF-κB pathway was activated, and p65 was transferred into the nucleus in high concentration of ACN group. In contrast, the activation of NF-κB was inhibited by NAC. Results of Western Blot showed that the expression of p65 protein was significantly increased and the expression of IκB protein was significantly decreased in high concentration of ACN group, while these changes were effectively abolished by NAC. Our study indicated that ACN-induced oxidative stress was able to activate NF-κB signaling pathway in rats testis.
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