The purpose of our study is to develop age-related phosphorylated tau (p-tau) inhibitors, for Alzheimer’s disease (AD). There are wide-ranging therapeutic molecules available in the market and tested for age-related p-tau inhibition to enhance phosphatase activity and microtubule stability in AD neurons. Until now there are no such small molecules claimed to show promising results to delay the disease process of AD. However, a recently developed molecule, DDQ, has been shown to reduce abnormal protein–protein interactions and protect neurons from mutant protein-induced toxicities in the disease process. In addition, DDQ reduced age- and Aβ-induced oxidative stress, mitochondrial dysfunction, and synaptic toxicity. To date, there are no published reports on the p-tau interaction of DDQ and Sirt3 upregulation with CREB-mediated mitophagy activation in AD neurons. In the current study, HT22 cells were transfected with mutant Tau (mTau) cDNA and treated with the novel molecule DDQ. Cell survival, immunoblotting, and immunofluorescence analysis were conducted to assess cell viability and synaptic and mitophagy proteins in treated and untreated cell groups. As expected, we found cell survival was decreased in mTau-HT22 cells when compared with control HT22 cells. However, cell survival was increased in DDQ-treated mTau-HT22 cells when compared with mTau HT22 cells. P-tau and total tau proteins were significantly reduced in DDQ-treated mTau-HT22 cells, and MAP2 levels were increased. Anti-aging proteins like Sirt3, and CREB levels were increased in DDQ-treated HT22 cells and also in mTau-HT22 cells treated DDQ. Mitophagy proteins were decreased in mTau-HT22 cells and these were increased in DDQ-treated mTau-HT22 cells. These observations strongly suggest that DDQ has anti-p-tau and anti-aging properties, via Sirt3 overexpression and increased mitophagy proteins. Our study findings may have implications for healthy aging to the development of p-tau targeted therapeutics in AD and tauopathies.

中文翻译：

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DDQ 抗衰老特性通过突变 tau HT22 神经元细胞线粒体自噬的改善而表达

我们研究的目的是开发与年龄相关的磷酸化 tau (p-tau) 抑制剂，用于治疗阿尔茨海默病 (AD)。市场上有多种治疗分子，并测试了与年龄相关的 p-tau 抑制作用，以增强 AD 神经元的磷酸酶活性和微管稳定性。到目前为止，还没有这样的小分子声称能够显示出延缓 AD 疾病进程的有希望的结果。然而，最近开发的一种分子 DDQ 已被证明可以减少异常的蛋白质-蛋白质相互作用，并保护神经元免受疾病过程中突变蛋白诱导的毒性。此外，DDQ 还可以减少年龄和 Aβ 诱导的氧化应激、线粒体功能障碍和突触毒性。迄今为止，尚无关于 AD 神经元中 DDQ 和 Sirt3 上调与 CREB ​​介导的线粒体自噬激活之间 p-tau 相互作用的报道。在目前的研究中，HT22 细胞被突变 Tau (mTau) cDNA 转染，并用新型分子 DDQ 处理。进行细胞存活、免疫印迹和免疫荧光分析，以评估处理和未处理的细胞组中的细胞活力以及突触和线粒体自噬蛋白。正如预期的那样，我们发现与对照 HT22 细胞相比，mTau-HT22 细胞的细胞存活率降低。然而，与 mTau HT22 细胞相比，DDQ 处理的 mTau-HT22 细胞的细胞存活率有所增加。DDQ 处理的 mTau-HT22 细胞中 P-tau 和总 tau 蛋白显着减少，MAP2 水平增加。DDQ 处理的 HT22 细胞以及 DDQ 处理的 mTau-HT22 细胞中抗衰老蛋白（如 Sirt3 和 CREB）水平升高。mTau-HT22 细胞中线粒体自噬蛋白减少，而 DDQ 处理的 mTau-HT22 细胞中线粒体自噬蛋白增加。这些观察结果强烈表明，DDQ 通过 Sirt3 过度表达和增加线粒体自噬蛋白而具有抗 p-tau 和抗衰老特性。我们的研究结果可能对健康老龄化以及 AD 和 tau 病的 p-tau 靶向疗法的开发产生影响。