Chen Cong-Ai, Li Chang-Xiang, Zhang Ze-Han, Xu Wen-Xiu, Liu Shu-Ling, Ni Wen-Chao, Wang Xue-Qian, Cheng Fa-Feng, Wang Qing-Guo
Dongzhimen Hospital Beijing University of Chinese Medicine, Beijing, 100700, China; Beijing University of Chinese Medicine, Beijing, 100029, China.
Beijing University of Chinese Medicine, Beijing, 100029, China.
J Ethnopharmacol. 2024 Jan 10;318(Pt A):116769. doi: 10.1016/j.jep.2023.116769. Epub 2023 Jul 1.
Qinzhizhudan Formula (QZZD) is composed of Scutellaria baicalensis Georgi (Huang Qin) extract, Gardenia jasminoides (Zhizi) extract and Suis Fellis Pulvis (Zhudanfen) (ratio of 4:5:6). This formula is optimized from Qingkailing (QKL) injection. Regarding brain injury, QZZD is protective. However, the mechanism by which QZZD treats vascular dementia (VD) has not been elucidated.
To ascertain QZZD's effect on the treatment of VD and further investigate the molecular mechanisms.
In this study, we screened the possible components and targets of QZZD against VD and microglia polarization using network pharmacology (NP), then an animal model of bilateral common carotid artery ligation method (2VO) was induced. Afterward, The Morris water maze was employed to evaluate cognitive ability, and pathological alterations in the CA1 area of the hippocampus were detected using HE and Nissl staining. To confirm the affect of QZZD on VD and its molecular mechanism, the contents of inflammatory factors IL-1β, TNF-α, IL-4, and IL-10 were performed to detect by ELISA, the phenotype polarization of microglia cells was detected by immunofluorescence staining, and the expressions of MyD88, p-IκBα and p-NF-κB p65 in brain tissue were detected by western blot.
A total of 112 active compounds and 363 common targets of QZZD, microglia polarization, and VD were identified, according to the NP analysis. 38 hub targets were screened out from the PPI network. GO analysis and KEGG pathway analysis showed that QZZD may regulate microglia polarization through anti-inflammatory mechanism such as Toll-like receptor signaling pathway and NF-κB signaling pathway. The further results showed that QZZD can alleviate the memory impairment induced by 2VO. QZZD profoundly rescued brain hippocampus neuronal damage and increased the number of neurons. These advantageous outcomes were linked to the control of microglia polarization. QZZD decreased M1 phenotypic marker expression while increasing M2 phenotypic marker expression. QZZD may controll the polarization of the M1 microglia by blocking the core part of Toll-like receptor signaling pathway, that is the MyD88/NF-κB signaling pathway, which reduced the neurotoxic effects of the microglia.
Here, we explored the anti-VD microglial polarization characteristic of QZZD for the first time and clarified its mechanisms. These findings will provide valuable clues for the discovery of anti-VD agents.
芩栀珠丹方（QZZD）由黄芩提取物、栀子提取物和猪胆汁粉（比例为4:5:6）组成。该方剂是由清开灵（QKL）注射液优化而来。在脑损伤方面，QZZD具有保护作用。然而，QZZD治疗血管性痴呆（VD）的机制尚未阐明。
确定QZZD对VD的治疗作用，并进一步探讨其分子机制。
在本研究中，我们使用网络药理学（NP）筛选QZZD针对VD和小胶质细胞极化的可能成分和靶点，然后采用双侧颈总动脉结扎法（2VO）诱导建立动物模型。随后，采用Morris水迷宫评估认知能力，并用HE和Nissl染色检测海马CA1区的病理改变。为了证实QZZD对VD的影响及其分子机制，采用ELISA检测炎症因子IL-1β、TNF-α、IL-4和IL-10的含量，通过免疫荧光染色检测小胶质细胞的表型极化，并用蛋白质免疫印迹法检测脑组织中MyD88、p-IκBα和p-NF-κB p65的表达。
根据NP分析，共鉴定出QZZD、小胶质细胞极化和VD的112种活性化合物和363个共同靶点。从蛋白质-蛋白质相互作用（PPI）网络中筛选出38个核心靶点。基因本体（GO）分析和京都基因与基因组百科全书（KEGG）通路分析表明，QZZD可能通过Toll样受体信号通路和NF-κB信号通路等抗炎机制调节小胶质细胞极化。进一步的结果表明，QZZD可以减轻2VO诱导的记忆损伤。QZZD能显著挽救脑海马神经元损伤并增加神经元数量。这些有利结果与小胶质细胞极化的调控有关。QZZD降低M1表型标记物的表达，同时增加M2表型标记物的表达。QZZD可能通过阻断Toll样受体信号通路的核心部分，即MyD88/NF-κB信号通路来控制M1小胶质细胞的极化，从而降低小胶质细胞的神经毒性作用。
在此，我们首次探讨了QZZD抗VD的小胶质细胞极化特性并阐明了其机制。这些发现将为抗VD药物的发现提供有价值的线索。
