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硒对砷中毒保护作用的实验研究
The Experiment Study of Protective Effect of Selenium on Arsenic Poisoning
【作者】 李萍;
【导师】 肖辉;
【作者基本信息】 新疆医科大学 , 营养与食品卫生学, 2011, 硕士
【摘要】 目的:通过体外、体内实验,研究亚硒酸钠(NaSeO2)单独及与磷酸钠(Na3PO4)联合应用对砷中毒的保护作用及其机制,为硒在砷中毒防治方面提供新的思路。方法:(1)体外实验:采用小鼠肝细胞原代培养技术,用一定浓度的亚砷酸钠(NaAsO2)处理小鼠肝细胞,同时分别加入不同剂量的亚硒酸钠、磷酸钠,通过噻唑蓝还原法(MTT),测定各组细胞吸光度值(OD),分析亚硒酸钠单独及与磷酸钠联合应用时对砷染毒小鼠肝细胞增殖的影响。(2)体内实验:SPF级昆明种(KM)小鼠70只,按体重随机分为7组:正常对照组、亚砷酸钠模型组、亚硒酸钠对照组、磷酸钠对照组、亚硒酸钠干预组、磷酸钠干预组、亚硒酸钠+磷酸钠干预组共7组。分别给以不同浓度的亚硒酸钠、磷酸钠进行干预,于60d结束实验并取材,采用黄嘌呤氧化酶法检测超氧化物歧化酶活性(SOD)、硫代巴比妥法检测丙二醛(MDA)含量、紫外分光法检测髓过氧化物酶(MPO)含量:高效液相色谱-氢化物发生原子荧光光谱法(HPLC-HGAFS)测定KM小鼠肝、肾组织中MMA(一甲砷酸)和DMA(二甲砷酸)含量。结果:(1)体外实验:低、中剂量(3μmol/L、5μmol/L、10μmol/L)的亚硒酸钠组OD值明显高于亚砷酸钠模型组(P<0.05);亚硒酸钠(3μmol/L)+磷酸钠(7.5μmol/L)干预组OD值明显高于其它干预组(P<0.05)(2)体内试验:亚砷酸钠模型组小鼠体重减轻,脏器系数升高,肝、肾中SOD、MPO含量低于正常对照组,MDA含量高于正常对照组(P<0.05);亚硒酸钠干预组体重明显高于亚砷酸钠模型组(P<0.05),脏器系数高于亚砷酸钠模型组(P<0.05),肝、肾中SOD、MPO含量明显高于亚砷酸钠模型组(P<0.05),MDA含量低于亚砷酸钠模型组(P<0.05);亚硒酸钠+磷酸钠干预组体重高于亚砷酸钠模型组(P<0.05),脏器系数低于亚砷酸钠模型组(P<0.05),小鼠肝、’肾中DMA、MMA较砷模型组降低(P<0.05)。亚硒酸钠十预组SOD活性、MPO、MDA、DMA、MMA含量与亚硒酸钠+磷酸钠干预组含量未见差异(P>0.05)结论:(1)亚硒酸钠低剂量(3μmol/L)对砷染毒小鼠肝细胞增殖有促进作用,而亚硒酸钠高剂量(25μmol/L)对砷染毒小鼠肝细胞增殖无促进作用,未见磷酸钠对亚硒酸钠促进砷染毒小鼠肝细胞增殖有协同作用。(2)亚硒酸钠对亚砷酸钠造成小鼠肝、肾的脂质过氧化损伤具有明显的保护作用,术见磷酸钠与亚硒酸钠联合应用对砷造成小鼠肝、肾的脂质过氧化损伤的协同效应。(3)硒对砷中毒具有一定的保护作用。
【Abstract】 Objective:By animal in vitro and in vivo, study of sodium selenite alone and with Trisodium phosphate combined application of arsenic poisoning of protection and its mechanism, for Sodium selenite on prevevtion and treatment of Arsenic poisoning provied new ideas. Methods:(1)In vitro, Using primary mouse liver cell culture technology, After processed cell by certain dosages of NaAsO2, different dosages of Sodium selenite and Trisodium phosphate were added to nutrient fluid, then through MTT, measured OD, Study influnce of sodium selenite alone or combined with Trisodium phosphate of arsenic exposure in mice on multiplication-inhibition of liver cells. (2)In vivo, SPF Kunming mice were 70, were randomly divided into seven groups: Control group, Arsenic in the model group, Sodium selenite control group, Trisodium phosphate control group, Sodium selenite Intervention group, Trisodium phosphate Intervention group, Sodium selenite+ Trisodium phosphate Intervention group, after 60 days, the animal were sacrificed and a series of biochemical indices were detected on organs. By xanthine oxidase activity of superoxide dismutase(SOD), thiobarbituric assay of malondialdehyde (MDA) content, ultraviolet spectrophotometry Detection of myeloperoxidase (MPO) content. (3)use(HPLC-HGAFS), determination of liver and kidney content of MMA and DMA. Results:(1)In vitro, low and medium dose(3μmol/L,5μmol/L,10μmol/L)Sodium selenite Intervention group OD was significantly higher than Arsenic in the model group(P<0.05). Sodium selenite+ Trisodium phosphate Intervention group OD was significantly higher than other groups(P <0.05). (2)In vivo, Arsenic exposure in mice have different degrees of weight loss, organ coefficient reduced; Arsenic group mouse liver, kidney SOD, MPO levels lower than the control group(P<0.05), MDA content was higher than the control group(P< 0.05); Sodium selenite Intervention group the weight Significantly higher than Arsenic in the model group(P< 0.05); MDA lower than Arsenic in the model group(P<0. 05);Sodium selenite+ Trisodium phosphate Intervention group the weight Significantly higher than Arsenic in the model group(P<0.05), Organ coefficient lower than Arsenic in the model group(P<0.05), DMA,MMA lower than Arsenic in the model group(P<0. 05); Sodium selenite Intervention group of Indicators and Sodium selenite+ Trisodium phosphate Intervention group of indicators No difference(P>0.05)Conclusions:(1)low doses of sodium selenite(3μmol/L)of arsenic exposure in mice can promote the proliferation of liver cells, High doses of sodium selenite(25μmol/L)of arsenic exposure on liver cell proliferation without promoting the effect. No synergy on Sodium selenite and Trisodium phosphate for arsenic exposure on liver cell proliferation. (2)Sodium selenite on sodium arsenite in mice liver and kidney lipid peroxidation damage has obvious protective effect. No synergy on Sodium selenite and Trisodium phosphate for Liver and kidney lipid peroxidation, (3)Sodium selenite on arsenic poisoning has a protective effect.
【Key words】 Sodium arsenite; Sodium selenite; Trisodium phosphate; Lipid peroxidation; experimental study;