ISSN 1674-3865  CN 21-1569/R
主管：国家卫生健康委员会
主办：中国医师协会
　　　辽宁省基础医学研究所
　　　辽宁中医药大学附属医院
中国中西医结合儿科学

中国中西医结合儿科学 ›› 2018, Vol. 10 ›› Issue (1): 1-5.doi: 10.3969/j.issn.1674-3865.2018.01.001

• 实验论著 •    下一篇

茸菖胶囊对无镁癫痫细胞模型海马神经元形态及存活状态的影响

戎萍,赵伟,张喜莲,王伟,张雪荣,马融   

  1. 300193 天津，天津中医药大学第一附属医院儿科（戎萍，张喜莲，马融），针灸研究所（王伟）；天津中医药大学2013级中医儿科学专业研究生（赵伟）；430061 武汉，湖北省中医院儿科（张雪荣）
  • 出版日期:2018-02-25 发布日期:2018-03-23
  • 通讯作者: 马融，E-mail：mr1974@163.com
  • 作者简介:戎萍（1978-），女，医学博士，副主任医师。研究方向：中医药防治小儿神经系统疾病
  • 基金资助:
    国家自然科学基金项目（81373691）

Impact of Rongchang capsule on hippocampal neuron morphology and survival status in magnesium-free epilepsy cell model

RONG Ping，ZHAO Wei，ZHANG Xilian，WANG Wei，ZHANG Xuerong，MA Rong   

  1. Department of Pediatrics,the First Teaching Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin 300193，China
  • Online:2018-02-25 Published:2018-03-23

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摘要：
目的
观察以益肾填精法为主的茸菖胶囊对无镁诱导的癫痫细胞模型海马神经元形态及存活状态的影响，探讨其对损伤神经元的保护作用及机制。
方法
原代培养新生SD大鼠海马神经元，建立无镁诱导的癫痫细胞模型，随机分为正常组（正常细胞外液）、模型组（无镁细胞外液）、对照组（无镁+3-甲基腺嘌呤）和5%、10%、20%茸菖胶囊组（无镁+不同浓度茸菖胶囊），采用倒置相差显微镜观察各组神经元形态学变化及凋亡情况，检测海马神经元活性及海马神经元损伤程度。
结果
（1）形态学变化：各时间点不同浓度茸菖胶囊组细胞较模型组海马神经元形态损伤减轻，胞体较完整，突起较清楚，存活细胞量多；（2）海马神经元活性：与正常组比较，模型组海马神经元细胞活力显著降低，差异有统计学意义（P<0.05）；与模型组比较，5%、10%、20%茸菖胶囊组显著提高无镁诱导的海马神经元的细胞活力，差异有统计学意义（P<0.05）；（3）海马神经元损伤程度：与模型组比较，5%、10%、20%茸菖胶囊组可显著减少无镁诱导所致的海马神经元内乳酸脱氢酶释放量，差异有统计学意义（P<0.05），并具有浓度依赖性；与正常组比较，模型组海马神经元乳酸脱氢酶释放量显著升高，差异有统计学意义（P<0.05）；5%、10%、20%茸菖胶囊组海马神经元内乳酸脱氢酶释放量随其浓度升高而逐渐降低。
结论
（1）茸菖胶囊能降低无镁诱导的癫痫细胞模型海马神经元乳酸脱氢酶释放量，维持细胞膜的完整性及线粒体功能，提高细胞活力并降低海马神经元凋亡率，起到保护海马神经元的作用。（2）茸菖胶囊提高对无镁诱导癫痫细胞模型神经元的活性，抑制海马神经元的凋亡，其神经保护作用可能与调控神经元自噬有关。

关键词: 癫痫, 无镁诱导, 海马神经元, 凋亡, 自噬, 茸菖胶囊, 大鼠

Abstract:
Objective
To observe the effect of Rongchang capsule characterized by tonifying kidney and replenishing essence on the morphology and survival status of magnesium-free epileptic models of hippocampal neuron， and discuss its protection effect on injured neuron and the probable mechanism.
Methods
Establish the magnesium-free epilepsy cell model by culturing the hippocampal neuron of newborn SD rats, which were randomly divided into normal group(normal extracellular fluid)， model group(magnesium-free extracellular fluid)， control group(magnesium-free+3-MA)， and the Rongchang capsule group of 5%,10% and 20% concentration(magnesium-free+Rongchang capsule of different concentration).The morphological changes and apoptosis of neutron cell of each group were observed through inverted phase contrast microscope; the activities of neutron cell were identified， and the injury degrees of neuron cell were determined.
Results
(1)Morphological changes： compared to the model group,the morphological damage of Chinese medicine cerebrospinal fluid group of different concentration were alleviated at each time point; the cell bodies were comparatively complete, the outgrowth was clear and there were more surviving cells.(2) Hippocampal neuron activity:compared with normal group， the activity of hippocampal neuron in model group was reduced remarkably(P<0.05); the activity of magnesium-free hippocampal neuron was greatly increased in the group processed with different concentration of Rongchang capsule compared with model group(P<0.05).(3)Hippocampus neuron damage degree：compared with model group， Rongchang capsule (low， medium， and high dose) group could significantly reduce the LDH release volume in magnesium-free hippocampal neuron， being concentration-dependent(P<0.05);  compared with normal group， the LDH release volume in hippocampal neuron of model group was significantly higher(P<0.05); the  LDH release volume in hippocampal neuronof Rongchang capsule(low， medium，and high dose) group decreased with the increase of concentration.
Conclusion
（1）Rongchang capsule can reduce the LDH release volume of magnesium-free epileptic model hippocampal neuron， keep the integrity of cell membrane and the function of mitochondria， increase the activity of neuron cells and decrease the rate of apoptosis， playing the role of protecting hippocampal neuron.（2）Rongchang capsule can increase the activity of neuron cells of magnesium-free epileptic models and depress the apoptosis of hippocampal neuron， and the protection mechanism might be related to regulating neuron's autophagy.

Key words: Magnesium-free induction, Epilepsy, Hippocampal neurons, Apoptosis, Autophagy, Rongchang capsule, Rat