慢性氟中毒大鼠脑组织中核因子E2相关因子2和胞质蛋白伴侣分子表达改变及其与氧化应激的关系  

Expression of NF-E2-related factor 2 and Kelch-like ECH-associated protein-1 in the brain of rats with chronic fluorosis and correlation with oxidative stress

作　　者：王娅[1] 董阳婷 魏娜[1] 官志忠 

机构地区：[1]贵州医科大学病理学教研室,贵阳550004 [2]贵州省医学分子生物学重点实验室

出　　处：《中华地方病学杂志》2015年第9期650-654,共5页Chinese Journal of Endemiology

基　　金：国家自然科学基金（81160335）;科技部国际合作重大项目（2013BA105B03）;贵州省科技厅国际合作项目（黔科合外G字[2011]7014号）

摘　　要：目的观察慢性氟中毒大鼠脑组织中核因子E2相关因子2（NF—E2-related factor 2，Nrf2）和胞质蛋白伴，吕分子（Kelch-like ECH-associated protein-1，Keapl）表达水平的改变，探讨其在慢性氟中毒脑损伤机制中的作用。方法SD大鼠90只，雌雄各半，按体质量采用随机数字表法分为3组（每组30只，雌雄各半），即对照组、染氟组（饮水氟含量50mg／L）、维生素E拮抗组（饮水氟含量50mg／L，同时给予维生素E5mg／kg灌胃），实验周期为10个月。染氟结束后，观察各组大鼠氟斑牙情况并收集24h尿样，处死大鼠，心尖取血并取后肢股骨及脑组织。采用氟离子选择电极法测定大鼠尿氟及骨氟含量；实时荧光定量PCR及蛋白印迹方法分别检测脑组织中Nrf2和Keapl的mRNA和蛋白表达；生化方法测定超氧化物歧化酶（SOD）和丙二醛（MDA）含量。结果染氟组大鼠出现不同程度的氟斑牙。各组间大鼠尿氟和骨氟比较差异均有统计学意义（F=6．87、182．87，P均〈0．05）。其中染氟组和维生素E拮抗组大鼠尿氟[（2．164-0．39）、（2．074-0．15）mg／L]和骨氟[（211．07±40．52）、（82．09±28．60）mg／kg]含量均高于对照组尿氟[（1．70±0．24）mg／L，P均〈0．05]和骨氟[（34．67±11．15）mg／kg，P均〈0．05]。各组间大鼠脑组织中Nrf2和Keapl的mRNA和蛋白表达水平、SOD活性、MDA含量比较差异均有统计学意义（F=654．33、432．87、447．45、398．88、68．34、68．34，P均〈0．05）。其中染氟组Nr￡2和KeaplmRNA[（320．18±6．83）％、（267．37±7．22）％]均高于对照组[（100．00±3．00）％、（100．00±2．75）％，P均〈0．05]，Nff2和Keapl蛋白表达水平[（283．28±6．89）％、（196．32±5．57）％]均高于对照组[（100．00±8．71）％、（100．00±9．23）％，P均〈0．05]；染氟组SOD活性[（22．10±2．10）μmol／kg]明显低于对照组[（35．05�Objective To observe the expression of NF-E2-related factor 2 （Nrf2） and Kelch-like ECH-associated protein-1 （Keapl） at mRNA and protein levels in the brain of rats with chronic fluorosis and to reveal the mechanism of brain damage induced by the factors. Methods Ninety SD rats were divided into three groups （30 rats in each group, half male half female） by the random number table method according to body weight. The control group was fed with normal tap-water, high-fluoride group with 50 mg/L fluoride （NaF） added in drinking water; and the high-fluoride plus vitamin E （Vit E）.group with the same dose of NaF as the high-fluoride group, but giving 5 mg/kg Vit E by intragastric administration. The experiment period was 10 months. The fluoride contents in urine and bone were detected by fluoride-ion selective electrode. The protein and mRNA levels of Nrf2 and Keapl in brain of rats were detected by Western blotting and quantitative real time PCR, respectively. The activity of superoxid dismutas （SOD） and the content of lipid peroxidation （MDA） were measured by biochemistry methods. Results Dental fluorosis was detected in high-fluoride group. The differences of fluoride contents in urine and bone were statistically significant between groups （F = 6.87, 182.87, all P 〈 0.05）. The urine fluoride [（2.16±0.39）, （2.07±0.15）mg/L] and bone fluoride [（211.07±40.52）, （82.09±28.60）mg/kg] in the high-fluoride and high-fluoride plus Vit E groups were higher than those of the control group [（1.70±0.24 ）mg/L, （34.67±11.15）mg/kg, all P 〈 0.05]. The differences of mRNA and protein levels of Nrf2 and Keapl in brains of rats, SOD activity, MDA content were statistically significant between groups （F = 654.33, 432.87, 447.45, 398.88, 68.34, 68.34, all P 〈 0.05）. The mRNA levels of Nrf2 and Keapl [（320.18±6.83）%, （267.37±7.22）%] were increased compared to those of control group [（100.00±3.00）%, （100.00± 2.75）%, all P 〈 0.05]; the protein level

关 键 词：氟中毒 牙 脑 氧化应激 

分 类 号：R599.1[医药卫生—内科学]