School of Pharmacy, Institute of Traditional Chinese Medicine and New Pharmacy Development, Guangdong Medical University, Dongguan, China.
Central Research Laboratory, Shenzhen Hospital of Integrated Traditional Chinese and Western Medicine, The Second People's Hospital of Bao'an Shenzhen (Group) , Shenzhen, China.
Neurotox Res. 2022 Aug;40(4):961-972. doi: 10.1007/s12640-022-00524-4. Epub 2022 Jun 14.
Alzheimer's disease (AD) causes progressive decline of memory and cognitive deficits. Because of its complicated pathogenesis, the prevention and therapy of AD remain an enormous challenge. It has been reported that catalpol possessed neuroprotective effects against AD. However, the involved mechanism still needs to be intensively studied. Therefore, the effects of catalpol on N2a/APP695swe cells and APP/PS1 mice were identified in the current study. Catalpol could improve cytotoxicity according to CCK-8 assay and ameliorate cellular morphological changes in N2a/APP695swe cells. Neuronal structural damage in the hippocampal CA1 region of APP/PS1 AD mice was improved according to HE staining and immunohistochemistry of NeuN. Meanwhile, catalpol administration ameliorated cognitive deficits confirmed by behavior performance of APP/PS1 mice. Hoechst 33,342 staining and Annexin V-FITC/PI double staining demonstrated that catalpol could reduce apoptosis in N2a/APP695swe cells. Likewise, TUNEL staining also manifested that catalpol significantly reduced apoptosis in hippocampal CA1 region of APP/PS1 mice. Catalpol administration also could improve mitochondrial functions indicated by the ameliorative mitochondrial morphology, the decreased ROS generation, and the increased MMP in N2a/APP695swe cells. Subsequently, catalpol restrained oligomerization of Aβ, verified by a reduced ThT fluorescence dose- and time-dependently. Additionally, both Aβ and Aβ aggregation were decreased in N2a/APP695swe cells and APP/PS1 mice administrated with catalpol confirmed by ELISA and western blot. Western blot also showed that catalpol facilitated the phosphorylation of AKT and GSK3β, and impeded the expression of BACE1 both in vivo and in vitro. Finally, a slight alteration in lactylation, 2-hydroxyisobutyrylation, and phosphorylation were found in N2a/APP695swe cells treated with catalpol. Together, these findings manifested that catalpol served a neuroprotective effect in AD and might be a novel and expecting prophylactic or curative candidate for AD or neurodegenerative diseases therapy.
阿尔茨海默病（AD）导致记忆和认知功能逐渐下降。由于其复杂的发病机制，AD 的预防和治疗仍然是一个巨大的挑战。据报道，梓醇对 AD 具有神经保护作用。然而，其涉及的机制仍需要深入研究。因此，本研究旨在确定梓醇对 N2a/APP695swe 细胞和 APP/PS1 小鼠的作用。CCK-8 检测结果表明，梓醇可以改善细胞毒性，改善 N2a/APP695swe 细胞的细胞形态变化。HE 染色和 NeuN 免疫组化结果表明，APP/PS1 AD 小鼠海马 CA1 区的神经元结构损伤得到改善。同时，行为学结果表明，梓醇可改善 APP/PS1 小鼠的认知功能障碍。Hoechst 33，342 染色和 Annexin V-FITC/PI 双染色结果表明，梓醇可减少 N2a/APP695swe 细胞的凋亡。同样，TUNEL 染色也表明，梓醇可显著减少 APP/PS1 小鼠海马 CA1 区的凋亡。梓醇给药还可以改善 N2a/APP695swe 细胞中线粒体的形态、减少 ROS 的产生和增加 MMP，从而改善线粒体功能。随后，梓醇通过减少 ThT 荧光剂量和时间依赖性地抑制 Aβ的寡聚化，验证了这一点。此外，ELISA 和 Western blot 结果表明，梓醇可减少 N2a/APP695swe 细胞和 APP/PS1 小鼠中的 Aβ和 Aβ 聚集。Western blot 结果还表明，梓醇可促进 AKT 和 GSK3β的磷酸化，同时减少体内和体外 BACE1 的表达。最后，在梓醇处理的 N2a/APP695swe 细胞中，发现了轻微的乳酰化、2-羟基异丁酰化和磷酸化改变。综上所述，这些发现表明梓醇在 AD 中具有神经保护作用，可能是 AD 或神经退行性疾病治疗的一种新型有前途的预防或治疗候选药物。
