Preliminary studies on effect and mechanism of serum with Bu Shen Decoction against excitatory neurotoxicity induced by glutamate

HE Wen-bin1; ZHANG Jun-long1; CHEN Nai-hong2; ZHANG Ling2; ZHU Hai-bo2

doi:10.13422/j.cnki.syfjx.2005.04.014

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Preliminary studies on effect and mechanism of serum with Bu Shen Decoction against excitatory neurotoxicity induced by glutamate

更新时间：2024-09-23

- Preliminary studies on effect and mechanism of serum with Bu Shen Decoction against excitatory neurotoxicity induced by glutamate
- Chinese Journal of Experimental Traditional Medical Formulae Issue 4, Pages: 28-32(2005)
- 作者机构：
  
  1.   山西中医学院
  2.   中国医学科学院药物研究所
  3.   中国医学科学院药物研究所太原030024
  4.   北京100050
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.2005.04.014
  CLC： R285.5
- Published：2005
- 稿件说明：
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[1]贺文彬,张俊龙,陈乃宏,张岭,朱海波.补肾方含药血清拮抗谷氨酸兴奋性神经毒作用及其机制的初步研究[J].中国实验方剂学杂志,2005(04):28-32.

HE Wen-bin1, ZHANG Jun-long1, CHEN Nai-hong2, et al. Preliminary studies on effect and mechanism of serum with Bu Shen Decoction against excitatory neurotoxicity induced by glutamate[J]. Chinese journal of experimental traditional medical formulae, 2005, (4): 28-32.
[1]贺文彬,张俊龙,陈乃宏,张岭,朱海波.补肾方含药血清拮抗谷氨酸兴奋性神经毒作用及其机制的初步研究[J].中国实验方剂学杂志,2005(04):28-32. DOI： 10.13422/j.cnki.syfjx.2005.04.014.

HE Wen-bin1, ZHANG Jun-long1, CHEN Nai-hong2, et al. Preliminary studies on effect and mechanism of serum with Bu Shen Decoction against excitatory neurotoxicity induced by glutamate[J]. Chinese journal of experimental traditional medical formulae, 2005, (4): 28-32. DOI： 10.13422/j.cnki.syfjx.2005.04.014.

摘要

目的:研究补肾复方含药血清（BS）对谷氨酸诱导凋亡PC12细胞的保护作用;并观察BS对谷氨酸诱导凋亡的PC12细胞胞浆内钙信号转导机制的影响。方法:采用MTT法测定谷氨酸致损伤后PC12细胞的存活力;采用荧光显微镜技术观察BS对谷氨酸所诱导PC12细胞凋亡的影响;以流式细胞术观察BS对谷氨酸诱导凋亡的PC12细胞胞浆内游离钙离子浓度的影响;运用蛋白免疫印迹（Westernblotting）技术观察BS对凋亡的PC12细胞CaMKⅡ的影响。结果:20%浓度BS含药血清可以拮抗谷氨酸的兴奋性神经毒作用

抑制PC12细胞凋亡;20%浓度BS含药血清能够拮抗谷氨酸诱导凋亡的PC12细胞胞浆内钙离子浓度的升高

抑制CaMKⅡ的磷酸化水平增高。结论:BS含药血清具有拮抗谷氨酸兴奋性神经毒的作用

其神经保护作用的机制可能与减轻钙超载

调控钙信号通路关键信号分子的磷酸化有关。

Abstract

Objectives:The antagonistic effects of serum with Bu Shen Decoction （BS） on apoptosis of PC12 cells induced by glutamate （Glu） were investigated in vitro. So were the effects of serum with BS on the calcium signal transduction system.Methods:The antagonistic action of the BS on neurotoxicity induced by Glu was observed with MTT method.The fluorescence microscope technique was employed to observe the effect of BS on apoptosis of PC12 cells induced by Glu. Its intracellular free calcium concentration was determined by flow cytometry.The Western-blotting technique was applied for observing the effect of BS on phosphorylation of CaMK Ⅱ in apoptotic PC12 cells.Results:The serum with BS in 20% concentration was discovered to be able to exert antagonistic effect on Glu-induced excitatory neurotoxicity and to inhibit the apoptosis of PC12 cells induced by Glu. Moreover

it was discovered that the increase in intracellular free calcium concentration and the excessive phosphorylation of CaMK Ⅱ during apoptosis of PC12 cells induced by Glu could be inhibited by serum with BS in 20% concentration.Conclusion:BS exert an antagonistic action on excitatory neurotoxicity induced by Glu.The mechanism of neuroprotective effect afforded by serum with BS might be related with relieving calcium overload and adjusting phosphorylation of the key signal molecules in the downstream of calcium ion signal transduction system.

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HE Wen-bin1

ZHANG Jun-long1

CHEN Nai-hong2

ZHANG Ling2

ZHU Hai-bo2

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