Xu Wenxiu, Ren Beida, Zhang Zehan, Chen Congai, Xu Tian, Liu Shuling, Ma Chongyang, Wang Xueqian, Wang Qingguo, Cheng Fafeng
School of Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing, China.
Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing, China.
Front Neurosci. 2022 Oct 20;16:943400. doi: 10.3389/fnins.2022.943400. eCollection 2022.
There is yet no effective drug for Alzheimer's disease (AD) which is one of the world's most common neurodegenerative diseases. The Qin-Zhi-Zhu-Dan Formula (QZZD) is derived from a widely used Chinese patent drug-Qing-Kai-Ling Injection. It consists of , , and . Recent study showed that QZZD and its effective components played important roles in anti-inflammation, antioxidative stress and preventing brain injury. It was noted that QZZD had protective effects on the brain, but the mechanism remained unclear. This study aims to investigate the mechanism of QZZD in the treatment of AD combining network pharmacology approach with experimental validation. In the network pharmacology analysis, a total of 15 active compounds of QZZD and 135 putative targets against AD were first obtained. Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were then applied to clarify the biological mechanism. The anti-inflammatory mechanism of QZZD was proved, and a synthetic pathway-TNFR1-ERK1/2-NF-κBp65 signaling pathway was obtained. On the basis of the above discoveries, we further validated the protective effects QZZD on neurons with an APP/PS1 double transgenic mouse model. Weight change of the mice was monitored to assess QZZD's influence on the digestive system; water maze experiment was used for evaluating the effects on spatial learning and memory; Western blotting and immunohistochemistry analysis were used to detect the predicted key proteins in network pharmacology analysis, including Aβ, IL-6, NF-κBp65, TNFR1, p-ERK1/2, and ERK1/2. We proved that QZZD could improve neuroinflammation and attenuate neuronal death without influencing the digestive system in APP/PS1 double transgenic mice with dementia. Combining animal pharmacodynamic experiments with network pharmacology analysis, we confirmed the importance of inflammation in pathogenesis of AD, clarified the pharmacodynamic characteristics of QZZD in treating AD, and proved its neuroprotective effects through the regulation of TNFR1-ERK1/2-NF-κBp65 signaling pathway, which might provide reference for studies on treatment of AD in the future.
阿尔茨海默病（AD）是世界上最常见的神经退行性疾病之一，目前尚无治疗该病的有效药物。芩栀珠丹方（QZZD）源自一种广泛使用的中成药——清开灵注射液。它由[此处原文缺失具体成分]组成。最近的研究表明，QZZD及其有效成分在抗炎、抗氧化应激和预防脑损伤方面发挥着重要作用。值得注意的是，QZZD对大脑有保护作用，但其机制尚不清楚。本研究旨在结合网络药理学方法和实验验证，探讨QZZD治疗AD的机制。在网络药理学分析中，首先获得了QZZD的15种活性化合物和135个针对AD的潜在靶点。然后应用基因本体（GO）富集分析和京都基因与基因组百科全书（KEGG）通路分析来阐明其生物学机制。证实了QZZD的抗炎机制，并获得了一条合成途径——TNFR1-ERK1/2-NF-κBp65信号通路。基于上述发现，我们进一步用APP/PS1双转基因小鼠模型验证了QZZD对神经元的保护作用。监测小鼠体重变化以评估QZZD对消化系统的影响；水迷宫实验用于评估对空间学习和记忆的影响；蛋白质免疫印迹法和免疫组织化学分析用于检测网络药理学分析中预测的关键蛋白，包括Aβ、IL-6、NF-κBp65、TNFR1、p-ERK1/2和ERK1/2。我们证明，在患有痴呆症的APP/PS1双转基因小鼠中，QZZD可以改善神经炎症并减轻神经元死亡，而不影响消化系统。将动物药效学实验与网络药理学分析相结合，我们证实了炎症在AD发病机制中的重要性，阐明了QZZD治疗AD的药效学特征，并证明了其通过调节TNFR1-ERK1/2-NF-κBp65信号通路发挥神经保护作用，这可能为未来AD治疗的研究提供参考。
