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 (12): 2270-2277.doi: 10.4103/1673-5374.285000

• 原著:退行性病与再生 • 上一篇    下一篇

酶消化阿胶可减少过氧化氢诱导神经元样PC12细胞死亡并加速淀粉样β蛋白的清除

  

  • 出版日期:2020-12-15 发布日期:2020-08-05
  • 基金资助:
    这项工作部分得到了日本科学促进会（JSPS）科学研究补助金(17K11813)和中国泰山工业专家计划(TSCY20170233）的资助

Enzyme-digested Colla Corii Asini (E’jiao) prevents hydrogen peroxide-induced cell death and accelerates amyloid beta clearance in neuronal-like PC12 cells

Li Xiao1, Feng Liao2, Ryoji Ide3, Tetsuro Horie4 , Yumei Fan2 , Chikako Saiki3 , Nobuhiko Miwa5   

  1. 1 Department of Pharmacology, Te Nippon Dental University, School of Life Dentistry at Tokyo, Tokyo, Japan
    2 National Engineering Research Center for Gelatin-based Traditional Chinese Medicine, Dong-E-E-Jiao Co. Ltd., Liaocheng, Shandong Province, China 
    3 Department of Physiology, Te Nippon Dental University, School of Life Dentistry at Tokyo, Tokyo, Japan
    4 Research Center, Te Nippon Dental University, School of Life Dentistry at Tokyo, Tokyo, Japan
    5 Department of Life Sciences, Prefectural University of Hiroshima, Hiroshima, Japan
  • Online:2020-12-15 Published:2020-08-05
  • Contact: Li Xiao, MD, PhD, xiaoli@tky.ndu.ac.jp.
  • Supported by:
    Tis work was supported in part by the Japan Society for the Promotion of Science (JSPS) Grant-in-Aid for Scientifc Research, No. 17K11813 (to LX) and TaiShan Industrial Experts Program of China No. TSCY20170233 (to FL).

PDF

177

摘要：

作为与衰老相关的退行性疾病，阿尔茨海默病的特征是淀粉样β蛋白（Aβ）沉积、氧化应激、炎症、功能障碍和胆碱能神经元丧失。中药阿胶可延缓衰老引起的神经细胞退行性变化，用于治疗与衰弱相关的疾病和抗衰老。实验进一步评估了酶消化的阿胶的抗氧化活性，神经细胞保护作用和对淀粉样β蛋白清除作用。氧自由基吸收能力活性测定表明，与来自猪、牛和冷水鱼皮肤的明胶相比，阿胶表现出最高的氧自由基吸收能力活性。扫描电镜观察的超微结构显示，2008，2013，2017年生产的阿胶和冷水鱼皮制成的明胶中，2008年产的阿胶呈现出最光滑的表面结构。实验进一步测试了2008年产的酶消化的阿胶和冷水鱼皮的酶消化明胶对过氧化氢处理的神经生长因子诱导 PC12分化的神经元样细胞死亡的保护作用。Presto blue分析显示，0.5 mg/mL的冷水鱼皮的酶消化明胶和酶消化的阿胶均可提高H2O2处理的神经元样PC12细胞活力。酶消化的阿胶的神经保护明显优于冷水鱼皮的酶消化明胶。乙酰胆碱酯酶分析表明，冷水鱼皮的酶消化明胶和酶消化的阿胶均可抑制神经元样PC12细胞的乙酰胆碱酯酶活性，分别达到未处理细胞的89.1％和74.5％。酶联免疫吸附反应和中性溶酶活性测定结果显示，酶消化的阿胶减少了淀粉样β蛋白1-42处理的神经元样PC12细胞中淀粉样β蛋白的积累，并增加了中性溶酶活性。这些结果表明，阿胶在阿尔茨海默病的预防和治疗方面有潜力。

orcid: 0000-0002-3053-4930 (Li Xiao)

关键词: 阿胶, 胶原蛋白, 抗氧化剂, PC12细胞, 阿尔茨海默病, 乙酰胆碱酯酶活性, 淀粉样β清除率, 神经保护

Abstract: As an aging-associated degenerative disease, Alzheimer’s disease is characterized by the deposition of am- yloid beta (Aβ), oxidative stress, infammation, dysfunction and loss of cholinergic neurons. Colla Corii Asini (CCA) is a traditional Chinese medicine which has been used for feebleness-related diseases and anti-aging. CCA might delay aging-induced degenerative changes in neurons. In the present study, we eval- uated antioxidant activity, cytoprotective efects, and Aβ removability of enzyme-digested Colla Corii Asini (CCAD). Oxygen radical absorbance capacity (ORAC) activity assay showed that, as compared to gelatins from the skin of porcine, bovine and cold water fsh, CCA exhibited the highest ORAC activity. Te ORAC activity of CCA and CCAD was increased gradually by the length of time in storage. Ultrastructure analysis by scanning electron microscopy showed that among CCA manufactured in 2008, 2013, 2017 and gelatin from cold water fsh skin, CCA manufactured in 2008 presented the smoothest surface structure. We fur- ther tested the protective efects of CCAD (manufactured in 2008) and enzyme-digested gelatin from cold water fsh skin (FGD) on hydrogen peroxide (H2O2)-induced cell death in nerve growth factor-diferenti- ated neuronal-like PC12 cells. Presto blue assay showed that both FGD and CCAD at 0.5 mg/mL increased cell viability in H2O2-treated neuronal-like PC12 cells. Te protection of CCAD was signifcantly superior to that of FGD. Acetylcholinesterase (AchE) assay showed that both FGD and CCAD inhibited AchE activ- ity in nerve growth factor-diferentiated neuronal-like PC12 cells to 89.1% and 74.5% of that in non-treated cells, respectively. Te data suggest that CCAD might be able to increase the neurotransmitter acetylcho- line. Although CCAD inhibited AchE activity in neuronal-like PC12 cells, CCAD prevented H2O2-induced abnormal deterioration of AchE. ELISA and neprilysin activity assay results indicated that CCAD reduced amyloid beta accumulation and increased neprilysin activity in Aβ1–42-treated neuronal-like PC12 cells, sug- gesting that CCAD can enhance Aβ clearance. Our results suggest that CCA might be useful for preventing and treating Alzheimer’s disease.

Key words: acetylcholinesterase activity, Alzheimer’s disease, amyloid beta clearance, antioxidant, Colla Corii Asini, collagen, neuroprotection, PC12 cells