氯化铜过度负荷对星形胶质细胞的氧化损伤机制  

Oxidative damage in the mechanism of copper toxicity in cultured astrocytes

作　　者：胡皓璐 王晓平[2] 赵艳玲[1] 

机构地区：[1]上海交通大学医学院附属第一人民医院老年科,200080 [2]上海交通大学附属第一人民医院神经内科

出　　处：《中华临床医师杂志（电子版）》2011年第11期3193-3200,共8页Chinese Journal of Clinicians(Electronic Edition)

基　　金：国家自然基金面上项目(81071065)

摘　　要：目的观察过量浓度Cu2+对原代培养的新生大鼠星形胶质细胞存活影响及探讨引起星形细胞抗氧化功能损伤的可能机制。方法原代培养的大鼠星形胶质细胞,分为低浓度(30μmol/L)、高浓度(100μmol/L)氯化铜(CuCl2)组和对照组,分别作用12、24、48、96、120h后,用CCK-8法检测星形细胞的存活率,比色法测定星形细胞内还原型谷胱甘肽(GSH)含量及谷胱甘肽还原酶(GR)的活性,Griess反应法测定NO分泌量。结果低浓度CuCl2可引起星形细胞的增殖,而高浓度组的存活率显著降低。高浓度CuCl2(100μmol/L)作用24h后细胞内GSH含量及GR活性显著低于对照组。作用星形细胞120h,NO分泌量较对照组显著增加。结论在病理情况下过度负荷的Cu2+显著损害星形胶质细胞的生长和增殖能力,通过降低GR活性及GSH的含量降低星形细胞的抗氧化功能。可能通过增加NO分泌加剧氧化应激并激活炎症途径。从而为临床治疗提供新的治疗分子靶点。Objective Copper is essential for several cellular processes in human being.However,excess copper is neuro-toxic and has been implicated in the pathogenesis of Wilson′s disease.This research is to investigate the effects of copper chloride on metabolism of the primary cultured astrocytes and possible mechanism of oxidative damage in cultured astrocytes.Methods Primary cultured astrocytes were exposed to 0,30,100 μmol/L CuCl2 respectively for 12 h,24 h,48 h,72 h,96 h,120 h,the change of the survival rate of astrocytes was analyzed (by CCK-8 assay).The activity of glutathione reductase(GR) and the levels of glutathione(GSH) in astrocytes were measured by colorimetric method,and nitric oxide (NO) production was measured with Griess method.Results There was an increased effect on cell growth with low concentration of copper chloride (P<0.05),but high concentration showed significantly inhibited (P<0.01).The GR activity and GSH amount decreased significantly after 24 h when astrocytes were treated with 100 μmol/L of copper chloride.NO secreted by astrocyte was increased significantly compared to that of the controls at 120 h.Conclusions The consequences suggested that excess copper affect the survival ratio on astrocytes.The decrease of GSH and the activity of GR may responsible for the disfunction of antioxidative of astrocyte.Astrocyte will product more NO to intensify oxidation and may play a role in the activation of inflammation.

关 键 词：铜 肝豆状核变性 帕金森病 星形细胞 氧化性应激 谷胱甘肽 

分 类 号：R965[医药卫生—药理学]