中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2019, Vol. 14 ›› Issue (5): 794-804.doi: 10.4103/1673-5374.249227

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

开心散的神经保护作用可改善东莨菪碱诱导的认知功能障碍

  

  • 出版日期:2019-05-15 发布日期:2019-05-15
  • 基金资助:

    国家自然科学基金(81473740，81673627，81673717)；广州科技创新委员会技术研究项目(2018050100)；广东省教育委员会特色创新基金(2016KTSCX011)；广州中医药大学高水平大学建设开放式抚育工程(2017年第34、118号)；中国新中医药临床试验技术平台项目(2012ZX09309-003，2008ZX09312-021)

Kai Xin San ameliorates scopolamine-induced cognitive dysfunctio

Yu-Min Xu 1, 2 , Xin-Chen Wang 3 , Ting-Ting Xu 1 , Hong-Ying Li 1 , Shang-Yan Hei 1 , Na-Chuan Luo 1 , Hong Wang 1 , Wei Zhao 1 , Shu-Huan Fang 1 , Yun-Bo Chen 1 , Li Guan 1 , Yong-Qi Fang 4 , Shi-Jie Zhang 1, 5 , Qi Wang 1, 6, Wei-Xiong Liang 1, 6   

  1. 1 Institute of Clinical Pharmacology, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong Province, China
    2 First Affiliated Hospital of Henan University of Chinese Medicine, Zhengzhou, Henan Province, China
    3 School of Food and Pharmaceutical Engineering, Zhaoqing University, Zhaoqing, Guangdong Province, China
    4 First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, Guangdong Province, China
    5 Department of Neurology, The First Affiliated Hospital, Guangzhou Medical University, Guangzhou, China
    6 Guangdong Provincial Hospital of Chinese Medicine, Guangzhou, Guangdong Province, China
  • Online:2019-05-15 Published:2019-05-15
  • Contact: Shi-Jie Zhang, MD, PhD, zsj19891122@gmail.com; Qi Wang, MD, wangqi@gzucm.edu.cn; Wei-Xiong Liang, MD, liangwx@yeah.net.
  • Supported by:

    This study was supported by the National Natural Science Foundation of China, No. 81473740, 81673627, 81673717 (to QW); Guangzhou Science Technology and Innovation Commission Technology Research Projects, China, No. 2018050100 (to QW); the Foundation for Characteristic Innovation of Educational Commission of Guangdong Province, China, Grant No. 2016KTSCX011 (to SHF); the Open Tending Project for Construction of High-Level University, Guangzhou University of Chinese Medicine, China, No. 34 and 118, 2017 (to SHF); the Technology Platform of Clinical Trials on New Traditional Medicine, China, No. 2012ZX09303009-003 (to WXL); the Technology Platform of Clinical Evaluation on New Traditional Medicine, China, No. 2008ZX09312-021 (to WXL).

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摘要：

中国传统中医药复方制剂开心散，主要成分包括人参、茯苓、石菖蒲及远志，调节认知功能的作用机制一直不甚明确。为此，实验设计选择广州中医药大学实验动物中心提供的8周龄健康 SPF 级雄性昆明小鼠72只，随机分为空白对照组、模型组、开心散低、中、高剂量组和阳性药对照组。除空白对照组和模型组予生理盐水外，开心散组分别以 0.7，1.4，2.8 g/kg/d开心散（中国广州中医药大学第一附属医院中药房）灌胃给药，阳性药对照组给予 3 mg/kg/d 的多奈哌齐（中国卫材药业有限公司），每日1次，治疗14 d。给药结束后第 8 日开始行为学测试，测试前 1 h灌胃给药，测试前半小时造模干预，除正常组用生理盐水代替外，其他各组小鼠予以东莨菪碱 3 mg/kg/d 腹腔注射。观察给药后测试开心散对模型小鼠空间学习记忆的影响，脑组织病理形态学变化，细胞凋亡因子、氧化应激损伤因子、突触相关蛋白的表达及胆碱能神经递质的表达变化。结果可见：(1) Morris水迷宫实验显示开心散可缩短模型小鼠的逃避潜伏期，增加目标象限停留时间和穿越平台次数；(2) Y迷宫实验中显示开心散可提高模型小鼠在迷宫臂之间的交替百分比；(3) Nissl和TUNEL染色显示开心散增加神经元内尼氏小体生成，抑制凋亡小体产生；(4) Western blot检测法发现开心散可上调抗凋亡蛋白Bcl-2的表达，抑制了促凋亡蛋白Bax的表达，可增加模型小鼠脑皮质和海马内突触后致密物95、突触素、脑源性神经营养因子蛋白的表达；(5)氧化应激相关因子检测显示开心散通过调节胆碱系统功能和改善氧化应激损伤以提高模型小鼠胆碱乙酰转移酶、乙酰胆碱、超氧化物歧化酶、谷胱甘肽过氧化物酶的活性和水平，抑制了乙酰胆碱酯酶、活性氧和丙二醛的活性和水平；(6)上述结果从不同途径证实了开心散通过抗细胞凋亡而减轻氧化应激损伤，调节突触相关蛋白和胆碱能神经递质从而发挥了神经保护和改善认知功能的作用。

orcid: 0000-0001-5399-7260 (Shi-Jie Zhang)
           0000-0002-3389-8529 (Qi Wang)
           0000-0002-9737-8549 (Wei-Xiong Liang)

关键词: 开心散, 认知功能障碍, 东莨菪碱氢溴酸盐, 神经保护, 氧化应激, 突触功能障碍, 细胞凋亡, 胆碱能系统功能障碍, 神经再生

Abstract:

Kai Xin San (KXS, containing ginseng, hoelen, polygala, and acorus), a traditional Chinese herbal compound, has been found to regulate cognitive dysfunction; however, its mechanism of action is still unclear. In this study, 72 specific-pathogen-free male Kunming mice aged 8 weeks were randomly divided into a vehicle control group, scopolamine group, low-dose KXS group, moderate-dose KXS group, high-dose KXS group, and positive control group. Except for the vehicle control group and scopolamine groups (which received physiological saline), the doses of KXS (0.7, 1.4 and 2.8 g/kg per day) and donepezil (3 mg/kg per day) were gastrointestinally administered once daily for 2 weeks. On day 8 after intragastric treatment, the behavioral tests were carried out. Scopolamine group and intervention groups received scopol¬amine 3 mg/kg per day through intraperitoneal injection. The effects of KXS on spatial learning and memory, pathological changes of brain tissue, expression of apoptosis factors, oxidative stress injury factors, synapse-associated protein, and cholinergic neurotransmitter were mea¬sured. The results confirmed the following. (1) KXS shortened the escape latency and increased residence time in the target quadrant and the number of platform crossings in the Morris water maze. (2) KXS increased the percentage of alternations between the labyrinth arms in the mice of KXS groups in the Y-maze. (3) Nissl and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling staining revealed that KXS promoted the production of Nissl bodies and inhibited the formation of apoptotic bodies. (4) Western blot assay showed that KXS up-regulated the expression of anti-apoptotic protein Bcl-2 and inhibited the expression of pro-apoptotic protein Bax. KXS up-regulated the expression of postsynaptic density 95, synaptophysin, and brain-derived neurotrophic factor in the cerebral cortex and hippocampus. (5) KXS increased the level and activity of choline acetyltransferase, acetylcholine, superoxide dismutase, and glutathione peroxidase, and re¬duced the level and activity of acetyl cholinesterase, reactive oxygen species, and malondialdehyde through acting on the cholinergic system and reducing oxidative stress damage. These results indicate that KXS plays a neuroprotective role and improves cognitive function through reducing apoptosis and oxidative stress, and regulating synapse-associated protein and cholinergic neurotransmitters.

Key words: Kai Xin San, cognitive dysfunction, scopolamine hydrobromide, neuroprotection, oxidative stress, synaptic dysfunction, apoptosis, cholinergic system dysfunction, donepezil, neural regeneration