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目的探讨维生素E(VE)和硒(Se)营养缺乏对食管肿瘤发生的影响及其氧化应激机制。方法 110只雄性F344大鼠随机分为3组:VE/Se缺乏组,VE/Se正常组和溶剂对照组。VE/Se缺乏组和VE/Se正常组动物皮下注射N-甲基苄基亚硝胺(NMBzA)染毒,染毒剂量为0.35 mg/kg BW,每周3次,连续5周。溶剂对照组动物皮下注射等体积的20%二甲基亚砜(DMSO)。VE/Se缺乏组给予低VE/Se饲料(VE 46U/kg,Se 0.05mg/kg),VE/Se正常组、溶剂对照组给予正常饲料(VE 80U/kg,Se 0.15mg/kg)。在实验第25周解剖动物,进行食管肿瘤肉眼观察及病理检查。动物血浆VE水平采用高效液相色谱法检测,Se水平采用荧光法检测。食管组织中细胞增殖和DNA氧化损伤分别采用5-溴脱氧尿嘧啶(BrdU)和8羟基脱氧鸟苷(8-OH-dG)免疫组织化学法检测。动物血浆、食管及肝脏谷胱甘肽过氧化物酶(GPX)、谷胱甘肽转移酶(GST)活性采用试剂盒法检测。结果动物血浆VE和Se水平VE/Se缺乏组显著低于VE/Se正常组,VE/Se正常组略低于溶剂对照组。VE/Se缺乏组动物食管肉眼可见肿瘤发生率、平均肿瘤数量及病理损伤数量均显著高于VE/Se正常组(P<0.05);与VE/Se正常组相比,VE/Se缺乏组动物食管组织细胞增殖水平、8-OH-dG水平显著升高(P<0.05);动物血浆、食管及肝脏GPX和GST活性显著降低(P<0.05)。结论维生素E和硒缺乏能够加快细胞增殖,显著促进NMBzA诱发的大鼠食管肿瘤的发生,机体氧化应激、DNA氧化损伤在食管癌变过程中具有重要作用。
Abstract：Objective To explore the effect of vitamin E(VE) and selenium(Se) deficiency on esophageal tumorigenesis and its oxidative stress mechanism.Methods 110 male F344 rats were randomly divided into 3 groups: VE/Se deficient group(group A),VE/Se normal group(group B) and control group(group C).Animals in group A and group B were subcutaneously injected with N-nitrosomethylbenzylamine(NMBzA) at the dosage of 0.35 mg/kg BW,3 times per week for 5 weeks.Control group was given the solvent 20% DMSO as negative control.Animals in group A were fed with low-VE/Se diet(46IU/kg VE,0.05mg/kg Se),animals in group B and C were fed with normal diet(80IU/kg VE,0.15mg/kg Se).Animals were sacrificed to perform macroscopic observation and pathologic inspection at the 25th week of the experiment.Plasma VE level was determined by high-performance liquid chromatography,and Se level was determined by fluorescence method.Cell proliferation and DNA damage in esophagus were detected through immunohistochemistry of BrdU and 8-OH-dG,respectively.Glutathione peroxidase(GPX) and glutathione transferase(GST) activities in plasma,esophagus and liver were analyzed by using test kits.Results VE/Se levels of group A were significantly lower than those of group B,the later were slightly lower than those of group C.Visible tumor incidence,tumor multiplicity and pathologic lesions in group A were all significantly more than those in group B.Comparing with group B,cell proliferation and 8-OH-dG levels were also significantly increased in group A.GPX and GST activities in plasma,esophagus and liver of group A were significantly higher than those of group B.Conclusion Vitamin E and selenium deficiency significantly promoted the NMBzA-induced esophageal tumorigenesis.Oxidative stress and DNA damage may be one of the critical routes by which NMBzA induces carcinogenesis in rat esophagus.
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基本信息:
DOI：10.19813/j.cnki.weishengyanjiu.2013.01.006
中图分类号:R735.1
引用信息:
[1]杨辉,李永宁,方瑾,等.维生素E和硒缺乏促进大鼠食管肿瘤发生及氧化应激机制的研究[J].卫生研究,2013,42(01):23-30.DOI:10.19813/j.cnki.weishengyanjiu.2013.01.006.
基金信息:
国家自然科学基金资助项目(No.30872110)
2013-01-30
2013-01-30