Lan Rui, Zhang Yong, Fu Xue-Qin, Wang Man-Man, Zou Xu-Huan, Wang Wei-Wei, Shen Xiao-Ming, Zhang Zhen-Qiang
The Department of Encephalopathy, The First Affiliated Hospital of Henan University of Chinese Medicine, Henan, 450000, China.
The Department of Traditional Chinese Medicine, The Third Affiliated Hospital of Zhengzhou University, Henan, 450000, China.
J Ethnopharmacol. 2025 Jan 31;340:119261. doi: 10.1016/j.jep.2024.119261. Epub 2024 Dec 17.
Ischemic stroke is an important cause of death and disability worldwide. Xiao-xu-ming Decoction (XXMD) is a classic prescription for the treatment of stroke patients, which has been widely used in China and has significant therapeutic effect, but the therapeutic target and mechanism are still unclear.
The current study aimed to investigate temporal alternation of synaptic damage and the protective effects of XXMD on synaptic damage following cerebral ischemia and reperfusion in vivo.
Adult male Sprague-Dawley (SD) rats subjected to 90 min of middle cerebral artery occlusion (MCAO) and subsequent reperfusion at various time points. Neurobehavioral function was assessed using modified neurological severity scores (mNSS), while pro-inflammatory cytokine levels were measured using ELISA kits. Histological assessment involved silver staining and Luxol Fast Blue (LFB) staining, and the ultrastructural alterations in neurons and synapses were examined using a transmission electron microscope (TEM). Golgi-cox staining was used to evaluate the density of dendritic spines. Levels of synapse-related proteins were quantified via immunofluorescence staining and Western blotting. Additionally, JAK2/STAT3 pathway related protein levels were assessed using Western blotting. In the second part, the rats were randomly divided into sham operation group, 24 h of reperfusion group, and XXMD group. The ultrastructural alterations and dendrite spine density of synapses were observed by TEM and Golgi-Cox staining respectively, and the expression levels of SYN, PSD95, GAP43, p-JAK2 and p-STAT3 were evaluated by WB.
Findings included deteriorated neurobehavioral function, increased release of IL-6, IL-1β, and TNFα, and time-dependent neuronal and synaptic damages during the initial phase of ischemia and reperfusion. At the ultrastructural level, neurons and synapses exhibited structural failure in the peri-infarct cortex. In addition, golgi-cox staining showed dendritic density in ischemic cortex significantly reduced after cerebral ischemia and reperfusion. Moreover, significant reductions in SYN, PSD95, and GAP43 expression levels, along with increases in p-JAK2 and p-STAT3 expression levels, were observed after cerebral ischemia and reperfusion. Meantime, XXMD significantly reduced synaptic impairment and up-regulated SYN, GAP43, PSD95 expression and down-regulated phosphorylation expression of JAK2 and STAT3 following MCAO and 24 h of reperfusion.
Collectively, these results indicates XXMD may play a neuroprotective role in reducing synaptic damage via JAK2/STAT3 signaling pathway.
缺血性中风是全球范围内死亡和残疾的重要原因。小续命汤（XXMD）是治疗中风患者的经典方剂，在中国已被广泛应用且具有显著的治疗效果，但其治疗靶点和机制仍不清楚。
本研究旨在探讨体内脑缺血再灌注后突触损伤的时间变化以及XXMD对突触损伤的保护作用。
成年雄性Sprague-Dawley（SD）大鼠接受90分钟的大脑中动脉闭塞（MCAO），随后在不同时间点进行再灌注。使用改良神经功能缺损评分（mNSS）评估神经行为功能，同时使用ELISA试剂盒测量促炎细胞因子水平。组织学评估包括银染色和Luxol Fast Blue（LFB）染色，并使用透射电子显微镜（TEM）检查神经元和突触的超微结构变化。使用高尔基-考克斯染色评估树突棘的密度。通过免疫荧光染色和蛋白质印迹法定量突触相关蛋白的水平。此外，使用蛋白质印迹法评估JAK2/STAT3通路相关蛋白的水平。在第二部分中，将大鼠随机分为假手术组、再灌注24小时组和XXMD组。分别通过TEM和高尔基-考克斯染色观察突触的超微结构变化和树突棘密度，并通过WB评估SYN、PSD95、GAP43、p-JAK2和p-STAT3的表达水平。
研究结果包括神经行为功能恶化、IL-6、IL-1β和TNFα释放增加，以及缺血再灌注初期神经元和突触的时间依赖性损伤。在超微结构水平上，梗死灶周围皮质的神经元和突触出现结构破坏。此外，高尔基-考克斯染色显示脑缺血再灌注后缺血皮质的树突密度显著降低。此外，脑缺血再灌注后观察到SYN、PSD95和GAP43表达水平显著降低，同时p-JAK2和p-STAT3表达水平升高。同时，XXMD在MCAO和再灌注24小时后显著减少突触损伤，上调SYN、GAP43、PSD95表达，并下调JAK2和STAT3的磷酸化表达。
总体而言，这些结果表明XXMD可能通过JAK2/STAT3信号通路在减少突触损伤方面发挥神经保护作用。
