Lai Bixuan, Wu Dan, Xiao Qidan, Wang Zhengyu, Niu Qixuan, Chen Qingjie, Long Qinghua, He Liling
Health Medical Center, Hubei Minzu University, Enshi, Hubei, 445000, PR China.
Health Medical Center, Hubei Minzu University, Enshi, Hubei, 445000, PR China; Hubei Key Laboratory of Diabetes and Angiopathy, Hubei University of Science and Technology, Xianning, Hubei, 437100, PR China.
J Ethnopharmacol. 2025 Feb 27;342:119424. doi: 10.1016/j.jep.2025.119424. Epub 2025 Jan 29.
Qiangji Decoction (QJD), a Chinese medicine, is widely used in Traditional Chinese Medicine to treat amnesia and Alzheimer's disease (AD), showing significant anti-AD effects. However, the precise mechanisms behind these effects are not well understood and require more research.
This study aims to elucidate the mechanisms by which QJD ameliorates neuronal damage, synaptic dysfunction, and mitochondrial impairment in AD through the regulation of ROCK2/Drp1-mediated mitochondrial dynamics.
UPLC-Q-TOF-MS/MS was used to identify active components in QJD extract. The study used SAMP8 mice for AD modeling and SAMR1 mice as controls. Cognitive function in SAMP8 mice was assessed with the Morris Water Maze after following treatment with QJD and the mitochondrial fission inhibitor Mdivi-1. Nissl and FJB staining evaluated QJD's effect on hippocampal injury. Synaptic integrity was examined with Golgi-Cox staining, transmission electron microscopy, and immunofluorescence. Mitochondrial function in hippocampal neurons was assessed using electron microscopy, JC-1 staining, and reagent kits. Western blot analyzed expression of proteins related to mitochondrial fission (ROCK2, Drp1, Fis1, Mff) and fusion (Mfn1, Mfn2, OPA1).
The analysis of QJD extract via UPLC-Q-TOF-MS/MS led to the identification of 46 active compounds. In SAMP8 mice, administration of QJD resulted in decreased escape latency, increased platform crossings, and extended duration in the target quadrant. Additionally, QJD exhibited neuroprotective effects on the hippocampus of SAMP8 mice, effectively preventing neuronal loss and damage. QJD also facilitated the extension and thickening of dendritic spines, enhanced the ultrastructure of hippocampal synapses, and upregulated synaptic function-related proteins, including PSD95 and SYN1. Furthermore, QJD ameliorated mitochondrial damage, improved mitochondrial membrane potential and ATP content, and reduced ROS expression in hippocampal neurons of SAMP8 mice. These effects were mediated through the downregulation of ROCK2, phosphorylated Drp1 (Ser616), Fis1, and Mff, as well as the upregulation of Mfn1, Mfn2, and OPA1.
QJD may reduce neuronal damage, synaptic dysfunction, and mitochondrial impairment in SAMP8 mice by regulating mitochondrial dynamics through the ROCK2/Drp1 pathway.
中药强肌汤（QJD）在传统中医中被广泛用于治疗失忆和阿尔茨海默病（AD），显示出显著的抗AD作用。然而，这些作用背后的确切机制尚不清楚，需要更多研究。
本研究旨在阐明QJD通过调节ROCK2/Drp1介导的线粒体动力学改善AD中神经元损伤、突触功能障碍和线粒体损伤的机制。
采用超高效液相色谱-四极杆飞行时间串联质谱（UPLC-Q-TOF-MS/MS）鉴定QJD提取物中的活性成分。本研究使用SAMP8小鼠建立AD模型，以SAMR1小鼠作为对照。用QJD和线粒体分裂抑制剂Mdivi-1处理后，通过莫里斯水迷宫评估SAMP8小鼠的认知功能。尼氏染色和FJB染色评估QJD对海马损伤的影响。用高尔基-考克斯染色、透射电子显微镜和免疫荧光检测突触完整性。使用电子显微镜、JC-1染色和试剂盒评估海马神经元的线粒体功能。蛋白质印迹法分析与线粒体分裂（ROCK2、Drp1、Fis1、Mff）和融合（Mfn1、Mfn2、OPA1）相关的蛋白质表达。
通过UPLC-Q-TOF-MS/MS对QJD提取物进行分析，鉴定出46种活性化合物。在SAMP8小鼠中，给予QJD可导致逃避潜伏期缩短、穿越平台次数增加以及在目标象限的停留时间延长。此外，QJD对SAMP8小鼠的海马体具有神经保护作用，有效预防神经元丢失和损伤。QJD还促进树突棘的延伸和增粗，增强海马突触的超微结构，并上调与突触功能相关的蛋白质，包括PSD95和SYN1。此外，QJD改善了线粒体损伤，提高了线粒体膜电位和ATP含量，并降低了SAMP8小鼠海马神经元中的活性氧表达。这些作用是通过下调ROCK2、磷酸化的Drp1（Ser616）、Fis1和Mff，以及上调Mfn1、Mfn2和OPA1介导的。
QJD可能通过ROCK2/Drp1途径调节线粒体动力学，减少SAMP8小鼠的神经元损伤、突触功能障碍和线粒体损伤。
