中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2020, Vol. 15 ›› Issue (4): 682-689.doi: 10.4103/1673-5374.266918

• 原著:退行性病与再生 • 上一篇    下一篇

传统中药益智提取物对过氧化氢诱导的PC12细胞损伤的保护作用

  


  • 出版日期:2020-04-15 发布日期:2020-05-28
  • 基金资助:
    国家自然科学基金（81574038）；中国广东省自然科学基金（2017A030313842，2017A050506007）; 深圳市基础研究项目技术研究基金（JCYJ20170412161254416）

Protective effects of organic extracts of Alpinia oxyphylla against hydrogen peroxide-induced cytotoxicity in PC12 cells

Li-Hong Duan1, 2, 3, 4, 5, Meng Li1, Chun-Bao Wang1, 2, 5, 6, 7, 8, Qing-Mei Wang3, Quan-Quan Liu1, 2, 5, 8, Wan-Feng Shang1, 2, 5, Ya-Jin Shen4, Zhuo-Hua Lin8, Tong-Yang Sun6, Zheng-Zhi Wu1, 2, 5, Ying-Hong Li1, 2, 5, Yu-Long Wang2, 5, Xun Luo9#br#   

  1. 1 Shenzhen Institute of Geriatrics, Shenzhen, Guangdong Province, China
    2 Department of Rehabilitation, the Second People’s Hospital of Shenzhen, Shenzhen, Guangdong Province, China
    3 Stroke Biological Recovery Laboratory, Spaulding Rehabilitation Hospital, Harvard Medical School, Charlestown, MA, USA
    4 Department of Mechanical and Biomedical Engineering, City University of Hong Kong, Hong Kong Special Administrative Region, China
    5 Department of Rehabilitation, the First Affiliated Hospital of Shenzhen University, Shenzhen, Guangdong Province, China
    6 School of Mechanical and Automotive Engineering, South China University of Technology, Guangzhou, Guangdong Province, China
    7 School of Mechanical Engineering, Guangxi University of Science and Technology, Liuzhou, Guangxi Zhuang Autonomous Region, China
    8 Mingkai Smart Medical Robot Co., Ltd., Shenzhen, Guangdong Province, China
    9 Kerry Rehabilitation Medicine Research Institute, Shenzhen, Guangdong Province, China
  • Online:2020-04-15 Published:2020-05-28
  • Contact: Chun-Bao Wang, PhD,chunbaowang@szu.edu.cn; Zheng-Zhi Wu, PhD,szwzz001@szu.edu.cn.
  • Supported by:
    This work was financially supported by the National Natural Science Foundation of China, No. 81574038 (to ZZW); the Natural Science Foundation of Guangdong Province of China, No. 2017A030313842 (to LHD); the Science and Technology Foundation of Guangdong Province of China, No. 2017A050506007 (to YHL); the Technology Research Foundation of Basic Research Project of Shenzhen City of China, No. JCYJ20170412161254416 (to ZZW).

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摘要：

益智是一种中国传统的草本植物，已广泛应用于神经保护、抗氧化和记忆改善等方面。然而，其活性成分的神经保护作用还不完全清楚。为了研究益智乙醇粗提物及其萃取部位是否具有神经保护作用。本研究将样本分为6组。将益智经95%的乙醇提取3次，获得第1，2，3次的3种提取液备用，作为第1，2，3组。并将第3次提取液的浓缩液悬浮于水后，采用不同的萃取剂(石油醚萃取部位，乙酸乙酯萃取部位，正丁醇萃取部位)萃取后所获得的3种萃取液作为另外3组进行实验。实验利用CCK-8检测和倒置显微镜观察细胞形态来测定上述各组分对过氧化氢诱导大鼠PC12细胞凋亡的神经保护作用，以确定上述6种组分中最具神经保护作用化合物的最佳剂量。结果发现：(1)益智由95%的乙醇提取3次的提取液(50，100，200 mg/mL)对H2O2引起大鼠PC12细胞氧化损伤的保护作用均呈剂量依赖性增加，最佳保护剂量为200 mg/mL；(2)益智萃取部位的乙酸乙酯部分(25，50，100 μg/mL)、石油醚部分(25，50，75 μg/mL)和正丁醇部分(100，150，200 μg/mL)对H2O2引起大鼠PC12细胞氧化损伤具有较好的保护作用，最佳有效浓度分别为50，25，200 mg/mL，其中乙酸乙酯萃取部位部分的保护作用尤为显著；(3) 上述结果证实益智的乙醇粗提物及其萃取部位对实验过氧化氢诱导的PC12细胞损伤有保护作用，为未来益智有效活性成分纯化研究提供了数据。

orcid: 0000-0002-8795-0142 (Chun-Bao Wang) 

         0000-0002-8819-609X (Zheng-Zhi Wu)

关键词: 益智, 传统草药, 乙醇粗提物, 萃取部位, 神经保护剂, 神经保护作用, 过氧化氢, 细胞凋亡, PC12细胞, 活性成分

Abstract: Alpinia oxyphylla, a traditional herb, is widely used for its neuroprotective, antioxidant and memory-improving effects. However, the neuroprotective mechanisms of action of its active ingredients are unclear. In this study, we investigated the neuroprotective effects of various organic extracts of Alpinia oxyphylla on PC12 cells exposed to hydrogen peroxide-induced oxidative injury in vitro. Alpinia oxyphylla was extracted three times with 95% ethanol (representing extracts 1–3). The third 95% ethanol extract was dried and resuspended in water, and then extracted successively with petroleum ether, ethyl acetate and n-butanol (representing extracts 4–6). The cell counting kit-8 assay and microscopy were used to evaluate cell viability and observe the morphology of PC12 cells. The protective effect of the three ethanol extracts (at tested concentrations of 50, 100 and 200 μg/mL) against cytotoxicity to PC12 cells increased in a concentration-dependent manner. The ethyl acetate, petroleum ether and n-butanol extracts (each tested at 100, 150 and 200 μg/mL) had neuroprotective effects as well. The optimum effective concentration ranged from 50–200 μg/mL, and the protective effect of the ethyl acetate extract was comparatively robust. These results demonstrate that organic extracts of Alpinia oxyphylla protect PC12 cells against apoptosis induced by hydrogen peroxide. Our findings should help identify the bioactive neuroprotective components in Alpinia oxyphylla.

Key words: active ingredients, Alpinia oxyphylla, apoptosis, ethanol crude extract, fraction, hydrogen peroxide, nerve regeneration, neuroprotective agent, neuroprotective effects, PC12 cells, traditional herb