Neural Regeneration Research ›› 2020, Vol. 15 ›› Issue (12): 2270-2277.doi: 10.4103/1673-5374.285000

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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).

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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