Guangdong Key Laboratory for Research and Development of Natural Drugs, School of Pharmacy, Guangdong Medical University, No. 1, Xincheng Road of Songshan Lake Science and Technology Industry Park, Dongguan, 523808, Guangdong, China.
Department of Pharmacy, Yuebei People's Hospital, Shaoguan, 512026, China.
Neurochem Res. 2020 May;45(5):1034-1044. doi: 10.1007/s11064-020-02978-4. Epub 2020 Feb 3.
Oxidative stress plays an important role in the pathogenesis of neurodegenerative diseases, including Alzheimer's disease and Parkinson's disease. Induction of endogenous antioxidants to act against oxidative stress-mediated neuronal damage seems to be a reasonable strategy for delaying the progression of such diseases. In this study, we investigated the neuroprotective effect of deuterium-depleted water (DDW) against HO-induced oxidative stress in differentiated PC12 cells and the possible signaling pathways involved. The differentiated PC12 cell line was pretreated with DDW containing different concentrations (50-100 ppm) of deuterium and then treated with HO to induce oxidative stress and neurotoxicity. We assessed cell survival, reactive oxygen species (ROS) generation, TUNEL assay, catalase (CAT), copper and zinc-containing superoxide dismutase (CuZn-SOD) and superoxide dismutase (SOD) activity and performed Western blot analysis to investigate the neuroprotective effect of DDW. The results indicated that DDW could attenuate HO-induced apoptosis, reduce ROS formation, and increase CAT, CuZn-SOD and SOD activity in HO-treated PC12 cells. Western blot analysis revealed that DDW treatment significantly increased the expression of p-Akt, Bcl-2 and GSK-3β. However, the protective effect of DDW on cell survival and the DDW-mediated increases in p-Akt, Bcl-2 and GSK-3β were abolished by pretreatment with the phosphatidylinositol-3-kinase (PI3K) inhibitor LY294002. In summary, DDW may protect differentiated PC12 cells against HO-induced oxidative stress through the PI3K/Akt signaling pathway.
氧化应激在神经退行性疾病（包括阿尔茨海默病和帕金森病）的发病机制中起着重要作用。诱导内源性抗氧化剂来对抗氧化应激介导的神经元损伤似乎是延缓这些疾病进展的合理策略。在这项研究中，我们研究了氘耗尽水（DDW）对分化的 PC12 细胞中 HO 诱导的氧化应激的神经保护作用及其可能涉及的信号通路。用含有不同浓度（50-100ppm）氘的 DDW 预处理分化的 PC12 细胞系，然后用 HO 处理以诱导氧化应激和神经毒性。我们评估了细胞存活率、活性氧（ROS）生成、TUNEL 测定、过氧化氢酶（CAT）、含铜和锌的超氧化物歧化酶（CuZn-SOD）和超氧化物歧化酶（SOD）活性，并进行 Western blot 分析以研究 DDW 的神经保护作用。结果表明，DDW 可减轻 HO 诱导的细胞凋亡，减少 ROS 形成，并增加 HO 处理的 PC12 细胞中的 CAT、CuZn-SOD 和 SOD 活性。Western blot 分析显示，DDW 处理可显著增加 p-Akt、Bcl-2 和 GSK-3β的表达。然而，PI3K 抑制剂 LY294002 预处理可消除 DDW 对细胞存活率的保护作用以及 DDW 介导的 p-Akt、Bcl-2 和 GSK-3β增加。综上所述，DDW 可能通过 PI3K/Akt 信号通路保护分化的 PC12 细胞免受 HO 诱导的氧化应激。
