中药健脾益肺方对EAAT2 表达的影响
作者:
作者单位:

(1.广州中医药大学第二临床医学院, 广州 510000;2.广东省中医院, 广州 510000)

作者简介:

通讯作者:

中图分类号:

R-33

基金项目:


Effect of Jianpi Yifei Prescription on the EAAT2 expression
Author:
Affiliation:

(1.The Second Clinical College of Guangzhou University of Chinese Medicine,Guangzhou 510006,China;2.Guangdong Provincial Hospital of Chinese Medicine,Guangzhou 510006)

Fund Project:

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    目的 探索健脾益肺方(JPYF)对谷氨酸转运体2(EAAT2)表达的影响及可能机制?方法 取新生24 h 内的SD 幼鼠,无菌条件下取出幼鼠大脑皮层星形胶质细胞,分离?提纯?原代培养以进行后期实验;调整适宜的细胞浓度分5 组种板,分别为A:谷氨酸组(250 μmoL/ L),B-D:JPYF 方(0. 25?0. 5?1 g/ kg)+谷氨酸组,E:正常组,以JPYF 方(终浓度为0. 25?0. 5?1 g/ kg)作用B-D 组24 h 后,谷氨酸250 μmoL/ L 作用A-D 组4 h 诱导建立体外培养原代星形胶质细胞损伤模型;采用噻唑蓝(MTT)法测定各组星形胶质细胞存活率,紫外比色法检测培养基上清谷氨酸浓度,细胞免疫荧光法(immunofluorescence)?Western blot 检测星形胶质细胞EAAT2 的表达量,IF 法检测原代脊髓神经元细胞存活情况?结果 JPYF 方可显著保护星形胶质细胞( P <0. 01),提高谷氨酸转运蛋白的表达( P <0. 05, P <0. 01),降低细胞外液谷氨酸浓度( P <0. 05, P <0. 01),减少神经元损害( P <0. 05, P <0. 01),在0. 25~1g/ L 的剂量上呈现一定的量效关系?结论 JPYF 方可减少谷氨酸对原代脊髓运动神经元细胞的损伤,其机制可能是JPYF 方提高EAAT2 的表达,降低细胞外液谷氨酸浓度,从而起到保护神经元的作用?

    Abstract:

    Objective To explore the effect of Jianpi Yifei prescription (JPYF) on the expression of glutamatetransporter 2 (EAAT2) and the possible mechanism behind this. Methods Neonatal Sprague-Dawley rats within 24 h ofbirth were used and and the astrocytes of cerebral cortex were isolated under aseptic conditions, then purified, and culturedfor later experiments. The appropriate cell concentration was adjusted in plates for five groups: A, glutamic acid group (250μmoL/ L), B-D, JPYF prescription (final concentrations 0. 25, 0. 5, and 1 g/ kg) + glutamic acid group; and E, normalgroup, After the B-D groups had been treated with JPYF prescription (final concentrations of 0. 25, 0. 5, and 1 g/ kg) for24 h, glutamate at 250 μmol/ L was applied to the A-D groups for 4 h to induce an in vitro primary astrocyte injury model.Cell viability was determined by MTT assay. Determination of glutamic acid concentration in culture medium was performedby ultraviolet colorimetry. The expression of glutamate transporter protein in astrocytes was detected by immunofluorescenceand western blotting. The injury of spinal cord neurons was detected by Immunofluorescence. Results JPYF Fangsignificantly protected the astrocytes ( P < 0. 01), improved the expression of glutamate transporters ( P < 0. 05, P < 0. 01),reduced glutamate concentrations ( P < 0. 05, P < 0. 01), protectd neurons ( P < 0. 05, P < 0. 01), and showed a certaindose-related effect within the range of 0. 25-1 g.L-1. Conclusions Within a certain dose range, JPYF Fang can reducethe damage of glutamate to neuronal cells, the mechanism of which may involve improving the expression of glutamate transporter and decreasing the concentration of glutamate in extracellular fluid, so as to protect neurons.

    参考文献
    相似文献
    引证文献
引用本文

李惠珍,任展能,李雅青,杨碧莹,杜宝新.中药健脾益肺方对EAAT2 表达的影响[J].中国比较医学杂志,2019,29(4):7~14.

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:2018-09-30
  • 最后修改日期:
  • 录用日期:
  • 在线发布日期: 2019-05-10
  • 出版日期:
自2024年1期开始,杂志参考文献改为中英文对照,具体格式要求可置下载中心查看!
关闭