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| 醒脑静注射液组分促进缺氧复氧胶质细胞清除胞外谷氨酸提高缺氧神经元活力的研究 |
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柴丽娟1,2,3, 徐耀1, 黄菊阳1,2, 李芮琳1, 郭虹1,2,3, 王少峡1,2, 胡利民1,2,3
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1.方剂学教育部重点实验室, 天津 301617;2.天津市中药药理学重点实验室, 天津 301617;3.天津中医药大学中医药研究院, 天津 301617
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| 摘要: |
| [目的]研究醒脑静注射液（XNJI）8种成分促进缺氧复氧（Hypo/Reox）星形胶质细胞对胞外高浓度谷氨酸的清除，及其条件培养液对缺氧后神经元活力的影响。[方法]体外培养大鼠星形胶质细胞，胶质纤维酸性蛋白（GFAP）免疫荧光鉴定。采用环磷酸腺苷（dBcAMP）和Hypo/Reox诱导胶质细胞活化并进行造模，Cell Counting Kit-8（CCK-8）测定细胞活力；谷氨酸试剂盒检测细胞培养液中谷氨酸浓度，并利用此条件培养液培养缺氧神经元，CCK-8法测神经元活力。[结果]胶质细胞体外培养成功，GFAP染色阳性。dBcAMP 0.062 5 mmol/L协同缺氧4 h复氧3 h（Hypo 4 h/Reox 3 h）处理后胶质细胞对胞外高浓度谷氨酸的清除能力显著降低。XNJI组分β-榄香烯、吉马酮、龙脑、冰片能显著提高受损胶质细胞对胞外谷氨酸的清除能力；XNJI组分吉马酮、龙脑、冰片处理的胶质细胞条件培养液能显著提高缺氧神经元的活力。[结论]XNJI组分能促进缺氧复氧胶质细胞对谷氨酸的清除并提高神经元活力。 |
| 关键词: 醒脑静注射液组分 胶质细胞 谷氨酸清除 缺氧复氧 神经元活力 |
| DOI：10.11656/j.issn.1672-1519.2019.03.19 |
| 分类号:R285.5 |
| 基金项目:国家自然科学基金资助项目（81573644）；国家中药标准化项目（ZYBZH-C-JS-35）；"十三五"期间天津市高等学校"创新团队培养计划"（TD13-5050）；天津市科技创新体系及条件平台建设计划（16PTSYJC00120）。 |
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| Xingnaojing injection components promote glutamate clearance in hypoxia/reoxygenation astrocytes and improve cell viability in hypoxia neurons |
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CHAI Lijuan1,2,3, XU Yao1, HUANG Juyang1,2, LI Ruilin1, GUO Hong1,2,3, WANG Shaoxia1,2, HU Limin1,2,3
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1.Key Laboratory of Pharmacology of Traditional Chinese Medical Formulae, Ministry of Education, Tianjin 301617, China;2.Tianjin Key Laboratory of Chinese Medicine Pharmacology, Tianjin 301617, China;3.Institute of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
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| Abstract: |
| [Objective] To study the effects of eight active components of Xingnaojing injection on glutamate transport in hypoxia/reoxygenation (Hypo/Reox) astrocytes in vitro and astrocyte-conditioned medium on the viability of Hypo/Reox neurons.[Methods] Astrocytes were cultured in vitro and identified by GFAP. Dibutyryl cyclic adenosine monophosphate (dBcAMP) and Hypo/Reox were used to induce astrocytes activation,cell viability was determined by CCK-8. Extracellular glutamate concentrations were measured by the Glutamate Kit. Neurons were cultured with astrocytes conditioned medium,and neuronal cell viability was determined by CCK-8.[Results] Astrocytes express the marker protein GFAP. dBcAMP 0.0625 mmol/L synergistic hypoxia 4 h plus reoxygenation 3 h (Hypo 4 h/Reox 3 h) can significantly reduce the ability of astrocytes to clear extracellular glutamate. The effective components of β-elemene,genistein,camphol and borneol can significantly increase the clearance ability of astrocytes in Hypo 4 h/Reox 3 h. At the same time,they also significantly improve neuronal viability cultured with astrocytes-conditioned medium after Hypo 4 h/Reox 3 h.[Conclusion] XNJI components promote glutamate clearance in hypoxia/reoxygenation astrocytes and improve cell viability in hypoxia neurons. |
| Key words: Xingnaojing injection components astrocytes glutamate clearance Hypo/Reox neuronal viability |
