莱菔硫烷与右美沙芬联合应用对甲基汞致大鼠神经毒性的影响  

Effects of sulforaphane combined with dextromethorphan on neurotoxicity induced by methylmercury in rats

作　　者：奉姝[1] 徐兆发[1] 王飞[1] 杨天瑶[1] 刘巍[1] 李乐慧[1] 邓宇[1] 徐斌[1] 

机构地区：[1]中国医科大学公共卫生学院环境卫生学教研室,辽宁沈阳110122

出　　处：《环境与健康杂志》2015年第8期663-667,共5页Journal of Environment and Health

基　　金：国家自然科学基金(81172631)

摘　　要：目的探讨甲基汞对大鼠神经毒性的影响及莱菔硫烷(sulforaphane,SFN)与右美沙芬(dextromethorphan,DM)联合应用的保护作用,为甲基汞中毒的防治提供理论依据。方法选取SPF级Wistar大鼠56只,按体重随机分成4组,每组14只,雌雄各半。其中对照组皮下及腹腔注射生理盐水;低、高剂量甲基汞染毒组皮下注射生理盐水,2 h后分别腹腔注射4、12μmol/kg氯化甲基汞;SFN+DM联合干预组皮下注射5.6μmol/kg SFN和13.5μmol/kg DM,2 h后腹腔注射12μmol/kg氯化甲基汞。每周一、三、五进行预处理,周一至周五染毒,连续4周。染毒结束后24 h分离大鼠脑皮质。采用冷原子吸收法测定大鼠脑皮质内汞含量,流式细胞仪测定细胞内Ca2+浓度、细胞凋亡率和活性氧(ROS)的含量,以试剂盒测定谷氨酸(Glu)、谷胱甘肽(GSH)含量及Na+-K+-ATP酶、Ca2+-ATP酶活力,以Real-time PCR和Western blotting法测定Nrf2、HO-1、γ-GCS、NR1、NR2A和NR2B的m RNA和蛋白表达。结果与对照组比较,低、高剂量染汞组大鼠的脑皮质汞、ROS、Ca2+含量及细胞凋亡率升高,GSH含量降低,Glu含量升高,Na+-K+-ATP酶和Ca2+-ATP酶活力降低,Nrf2、HO-1、γ-GCS m RNA和蛋白表达量升高,NR1、NR2A和NR2B的m RNA和蛋白表达量下降,差异均有统计学意义(P<0.05或P<0.01)。与高剂量染汞组比较,SFN+DM联合干预组大鼠脑皮质汞含量差异无统计学意义(P>0.05),ROS、Ca2+含量及细胞凋亡率降低,GSH含量升高,Glu含量降低,Na+-K+-ATP酶和Ca2+-ATP酶活力升高,HO-1、γ-GCS、NR1、NR2A的m RNA和蛋白表达量升高,差异均有统计学意义(P<0.05或P<0.01)。结论甲基汞可引发大鼠神经毒性,SFN与DM联合应用可有效拮抗甲基汞所致神经毒性。Objective To explore the effects of sulforaphane combined with dextromethorphan on toxic effect induced by methylmercury in rat cerebral cortex. Methods Fifty-six Wistar rats were randomly divided into four groups by weight. The first group was control group, the second and third groups were the low-dose and high-dose CH3 HgCl administration groups,the fourth group was the sulforaphane（SFN） combined with dextromethorphan（DM） pretreatment group. The rats in the first,second and third groups were subcutaneously injected with 0.9% saline solution, while the rats in the fourth group was given 5.6 μmol/kg SFN combined with 13.5 μmol/kg DM in advance. Two hours later, the rats in the first group was intraperitoneally injected with 0.9% saline solution, while the rats in the second, third and fourth groups were respectively intraperitoneally injected daily with CH3 HgCl in a single dose of 4, 12 and 12 μmol/kg body weight. The pretreatment and CH3 HgCl treatment lasted for four weeks. Twenty-four hours after the last treatment, the Hg levels, apoptosis rate, intracellular ROS, calcium concentrations, contents of Glu and GSH, activities of Na＋-K＋-ATPase and Ca2 ＋-ATPase, and the m RNA and protein expressions of Nrf2, HO-1, γ-GCS, NR1, NR2 A and NR2 B were all determined. Results Compared with control group, the mercury content, apoptosis rate, intracellular ROS, calcium concentrations, Glu contents, the m RNA and protein expressions of Nrf2, HO-1 and γ-GCS were all increased significantly, while GSH contents, activities of Na＋-K＋-ATPase and Ca2＋-ATPase,and the m RNA and protein expressions of NR1, NR2 A and NR2 B were decreased in methylmercury-exposed group in a dosedependent manner,and the difference were statistically significant（P〈0.05 or P〈0.01）. Compared with the high methylmercury group, the apoptosis rate, ROS levels,the content of Glu and Ca2＋reduced respectively,the levels of GSH, Na＋-K＋-ATPase and Ca2 ＋-ATPase increased significantly,the m RNA and protein expressions of

关 键 词：甲基汞 神经毒性 莱菔硫烷 右美沙芬 联合应用 

分 类 号：R994.6[医药卫生—毒理学]