李氏5号水提液在兴奋性氨基酸损伤神经元中的保护作用

南方医科大学学报 ›› 2005, Vol. 25 ›› Issue (10): 1295-1298.

李氏5号水提液在兴奋性氨基酸损伤神经元中的保护作用

胡德辉¹, 杨建明², 常全忠³, 李生海⁴, 李子中⁴

1. 南方医科大学生理教研室, 广东, 广州, 510515;
2. 南方医科大学神经生物教研室, 广东, 广州, 510515;
3. 新乡医学院生理教研室, 河南, 新乡, 453003;
4. 高明脑病医疗医药研究院, 广东, 高明, 528500

出版日期:2005-10-20 发布日期:2005-10-20
基金资助:
收稿日期:2005-3-19。
作者简介:胡德辉(1965- ),男,博士,副教授,电话:020-61648603

Protective effects of liquid extract of Lishi No.5 formula against excitatory amino acid damage of neurons

HU De-Hui¹, YANG Jian-ming², CHANG Quan-zhong³, LI Sheng-hai⁴, LI Zi-zhong⁴

1. 南方医科大学生理教研室, 广东, 广州, 510515;
2. 南方医科大学神经生物教研室, 广东, 广州, 510515;
3. 新乡医学院生理教研室, 河南, 新乡, 453003;
4. 高明脑病医疗医药研究院, 广东, 高明, 528500

Online:2005-10-20 Published:2005-10-20

摘要/Abstract

摘要： 目的探讨李氏5号水提液在兴奋性氨基酸性神经元损伤中的保护作用。方法 MTT法观察不同干扰因素作用下细胞成活率和用Hoechest333258染色法荧光显微镜下观察细胞凋亡的比例;借助共聚焦显微镜钙成像和钙荧光指示剂Fluo-3/AM测细胞内钙浓度。结果 300μmol/LNMDA+5μmol/L甘氨酸作用10min后换正常培养液培养18h,细胞死亡率比正常对照组高(54.64±5.76)%;用李氏5号水提液(终浓度为0.5mg/ml)预处理3h后,加300μmol/LNMDA+5μmol/L甘氨酸作用10min,换正常培养液同时加李氏5号水提液继续培养18h,细胞死亡率明显降低。10μmol/LNMDA可明显增加海马神经元胞质游离钙浓度。李氏5号水提液本身可轻度增加胞质游离钙浓度(与正常组比较差异不显著),但明显抑制NMDA导致的胞质游离钙浓度的升高;NMDA阻断剂MK-801明显拮抗NMDA导致胞质游离钙升高的效应,但李氏5号水提液预处理后,MK-801拮抗NMDA的效应显著降低。结论李氏5号水提液可能通过拮抗兴奋性氨基酸所致的细胞内钙超载而明显保护神经元。

Abstract: Objective To investigate the effect of liquid extract of Lishi No.5 formula in protecting the neurons from excitatory amino acid injury. Methods Cultured neonatal SD rat hippocampal neurons were treated with 300 μmol/L NMDA and 5 μmol/L glycine for 10 min, and the conditioned culture medium was replaced by normal culture medium. After cell culture for 18 h, MTT assay and Hoechst33258 staining were performed to examine the cell survival rate and cell apoptosis, respectively. Intracellular free calcium concentration was assayed by image analysis of the calcium signals with confocal microscope and Fluo-3/AM indicator. Results The survival rate of the cultured cell was significanty decreased in response to treatment with 300 μmol/L NMDA and 5 μmol/L glycine, but the effects were obviously reversed by treatment with 0.5 mg/ml liquid extract from Lishi No.5 formula. Intracellular free calcium concentration was significantly increased by 10 μmol/L NMDA, which was remarkably inhibited by the liquid extract. The effects of NMDA on intracellular free calcium was abolished by pretreatment of the cells with MK-801 for 10 min, whereas the liquid extract significantly lowered the antagonizing effect of MK-801. Conclusion Lishi No.5 formula protects the neurons from toxicity by excitatory amino acids possibly by alleviating intracellular free calcium overload induced by these amino acids.

中图分类号:

胡德辉¹, 杨建明², 常全忠³, 李生海⁴, 李子中⁴. 李氏5号水提液在兴奋性氨基酸损伤神经元中的保护作用[J]. 南方医科大学学报, 2005, 25(10): 1295-1298.

HU De-Hui¹, YANG Jian-ming², CHANG Quan-zhong³, LI Sheng-hai⁴, LI Zi-zhong⁴. Protective effects of liquid extract of Lishi No.5 formula against excitatory amino acid damage of neurons[J]. Journal of Southern Medical University, 2005, 25(10): 1295-1298.

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