Xiao Li, Liao Feng, Ide Ryoji, Horie Tetsuro, Fan Yumei, Saiki Chikako, Miwa Nobuhiko
Department of Pharmacology, The Nippon Dental University, School of Life Dentistry at Tokyo, Tokyo, Japan.
National Engineering Research Center for Gelatin-based Traditional Chinese Medicine, Dong-E-E-Jiao Co. Ltd., Liaocheng, Shandong Province, China.
Neural Regen Res. 2020 Dec;15(12):2270-2272. doi: 10.4103/1673-5374.285000.
As an aging-associated degenerative disease, Alzheimer's disease is characterized by the deposition of amyloid beta (Aβ), oxidative stress, inflammation, 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 evaluated antioxidant activity, cytoprotective effects, 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 fish, CCA exhibited the highest ORAC activity. The 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 fish skin, CCA manufactured in 2008 presented the smoothest surface structure. We further tested the protective effects of CCAD (manufactured in 2008) and enzyme-digested gelatin from cold water fish skin (FGD) on hydrogen peroxide (HO)-induced cell death in nerve growth factor-differentiated neuronal-like PC12 cells. Presto blue assay showed that both FGD and CCAD at 0.5 mg/mL increased cell viability in HO-treated neuronal-like PC12 cells. The protection of CCAD was significantly superior to that of FGD. Acetylcholinesterase (AchE) assay showed that both FGD and CCAD inhibited AchE activity in nerve growth factor-differentiated neuronal-like PC12 cells to 89.1% and 74.5% of that in non-treated cells, respectively. The data suggest that CCAD might be able to increase the neurotransmitter acetylcholine. Although CCAD inhibited AchE activity in neuronal-like PC12 cells, CCAD prevented HO-induced abnormal deterioration of AchE. ELISA and neprilysin activity assay results indicated that CCAD reduced amyloid beta accumulation and increased neprilysin activity in Aβ-treated neuronal-like PC12 cells, suggesting that CCAD can enhance Aβ clearance. Our results suggest that CCA might be useful for preventing and treating Alzheimer's disease.
作为一种与衰老相关的退行性疾病，阿尔茨海默病的特征在于β淀粉样蛋白（Aβ）沉积、氧化应激、炎症、胆碱能神经元功能障碍和丧失。阿胶是一种传统中药，已用于治疗虚弱相关疾病和抗衰老。阿胶可能会延缓衰老引起的神经元退行性变化。在本研究中，我们评估了酶解阿胶（CCAD）的抗氧化活性、细胞保护作用和Aβ清除能力。氧自由基吸收能力（ORAC）活性测定表明，与猪皮、牛皮和冷水鱼皮明胶相比，阿胶表现出最高的ORAC活性。阿胶和CCAD的ORAC活性随储存时间的延长而逐渐增加。扫描电子显微镜超微结构分析表明，在2008年、2013年、2017年生产的阿胶和冷水鱼皮明胶中，2008年生产的阿胶表面结构最光滑。我们进一步测试了2008年生产的CCAD和冷水鱼皮酶解明胶（FGD）对神经生长因子分化的神经元样PC12细胞中过氧化氢（HO）诱导的细胞死亡的保护作用。Presto blue测定表明，0.5mg/mL的FGD和CCAD均增加了HO处理的神经元样PC12细胞的活力。CCAD的保护作用明显优于FGD。乙酰胆碱酯酶（AchE）测定表明，FGD和CCAD分别将神经生长因子分化的神经元样PC12细胞中的AchE活性抑制至未处理细胞的89.1%和74.5%。数据表明CCAD可能能够增加神经递质乙酰胆碱。虽然CCAD抑制了神经元样PC12细胞中的AchE活性，但CCAD可防止HO诱导的AchE异常降解。ELISA和中性内肽酶活性测定结果表明，CCAD减少了Aβ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内肽酶活性，表明CCAD可增强Aβ清除。我们的结果表明阿胶可能对预防和治疗阿尔茨海默病有用。
