毛蕊花苷对血红素氧化酶-1过表达大鼠原代星形胶质细胞铁沉积的影响

郭欢，王涛，王海涛，张永东，郭红云，李永辉，闵建平，刘元强，赵勤，苏海翔

中国药学杂志 ›› 2017, Vol. 52 ›› Issue (2) : 109-114.

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中国药学杂志
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中国药学杂志 ›› 2017, Vol. 52 ›› Issue (2) : 109-114. DOI: 10.11669/cpj.2017.02.006
论著

毛蕊花苷对血红素氧化酶-1过表达大鼠原代星形胶质细胞铁沉积的影响

  • 郭欢，王涛，王海涛，张永东，郭红云，李永辉，闵建平，刘元强，赵勤，苏海翔*
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Reduction of Iron Deposition by Verbascoside in HO-1 Overexpression Rats Astrocytes

  • GUO Huan, WANG Tao, WANG Hai-tao, ZHANG Yong-dong, GUO Hong-yun, LI Yong-hui, MIN Jian-ping, LIU Yuan-qiang, ZHAO Qin, SU Hai-xiang *
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摘要

目的 观察毛蕊花苷( verbascoside )对过表达血红素氧化酶-1(heme oxygenase-1，HO-1)的大鼠原代星形胶质细胞铁离子沉积的影响，寻找抗阿尔兹海默病的新药物。方法 体外培养和纯化大鼠原代星形胶质细胞，构建转染HO-1基因的星形胶质细胞模型，用CCK-8法检测细胞损伤，Western blotting法检测 HO-1 蛋白的表达，普鲁士蓝染色法检测细胞铁离子沉积。结果 95%以上的细胞呈胶质纤维酸性蛋白(GFAP)阳性，表明成功培养出星形胶质细胞;毛蕊花苷对 HO-1基因转染后的原代星形胶质细胞具有不同程度的增殖抑制效应，抑制率与药物浓度呈正相关;转染HO-1基因入星形胶质细胞后，HO-1蛋白表达增加6.86倍;在施加毛蕊花苷干预后，20、40、60 μmol·L-1均能不同程度地降低HO-1蛋白的表达，同时不同程度地减少由过表达HO-1导致的铁离子沉积(20 μmol·L-1降低 47.69%，40 μmol·L-1降低 51.63%)。结论 毛蕊花苷可以抑制过表达 HO-1 大鼠原代星形胶质细胞的铁离子沉积，可能成为针对“铁-氧化损伤”途径的抗阿尔兹海默病的新药物。

Abstract

OBJECTIVE To investigate the effect of verbascoside on iron deposition of astrocyte cell overexpressing heme oxygenase-1 gene, and to seek new drug in treating Alzheimer′s disease. METHODS Primary cultured astrocyte cells of rats were cultured and purified in vitro. With gene recombination and transfer techniques, transfect HO-1 gene into primary cultured astrocyte cells.HO-1 transfected astrocyte were then treated by different concentrations of verbascoside.Proliferation of cell was measured by CCK-8 assay and expression of HO-1 protein was analyzed by Western blotting. Iron deposition was also visualized by Perl′s Prussian blue stain. RESULTS Immunofluorescence staining showed that most of the cells were GFAP-postitive and the purity was over 95%. The expression of HO-1 protein was increased in HO-1 injury model of astrocyte. Treatment of verbascoside with variable concentration caused the decrease of HO-1 protein expression more or less and reduction of iron deposition in transfected cells(20 μmol·L-1reducing 47.69%,40 μmol·L-1 reducing 51.63%). CONCLUSION Verbascoside could enhance astrocyte cell resistance against injury induced by overexpression of heme oxygenase-1 gene.

关键词

大鼠原代星形胶质细胞 / 血红素氧化酶-1 / 铁离子沉积 / 阿尔兹海默病 / 毛蕊花苷

Key words

astrocyte / heme oxygenase-1 / iron deposition / Alzheimer′s disease / verbascoside

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郭欢，王涛，王海涛，张永东，郭红云，李永辉，闵建平，刘元强，赵勤，苏海翔. 毛蕊花苷对血红素氧化酶-1过表达大鼠原代星形胶质细胞铁沉积的影响[J]. 中国药学杂志, 2017, 52(2): 109-114 https://doi.org/10.11669/cpj.2017.02.006
GUO Huan, WANG Tao, WANG Hai-tao, ZHANG Yong-dong, GUO Hong-yun, LI Yong-hui, MIN Jian-ping, LIU Yuan-qiang, ZHAO Qin, SU Hai-xiang. Reduction of Iron Deposition by Verbascoside in HO-1 Overexpression Rats Astrocytes[J]. Chinese Pharmaceutical Journal, 2017, 52(2): 109-114 https://doi.org/10.11669/cpj.2017.02.006
中图分类号： R965   

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基金

甘肃省中医药管理局科学技术研究课题(GZK-2010-Z7);甘肃省卫生行业科研计划项目(GSWSKY-2015-25);甘肃省卫生行业科研计划管理项目(GWGL2013-12)
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