Yang Tong, Liu Xiaolu, Zhou Yue, Du Lipeng, Fu Yang, Luo Yanan, Zhang Wenli, Feng Zhitao, Ge Jinwen, Mei Zhigang
Key Laboratory of Hunan Province for Integrated Traditional Chinese and Western Medicine on Prevention and Treatment of Cardio-Cerebral Diseases, College of Integrated Traditional Chinese and Western Medicine, Hunan University of Chinese Medicine, Changsha, 410208, Hunan, China.
Third-Grade Pharmacological Laboratory on Chinese Medicine Approved by State Administration of Traditional Chinese Medicine, College of Medicine and Health Sciences, China Three Gorges University, Yichang, 443002, Hubei, China; State Key Laboratory of Natural Medicines and School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing, 210009, Jiangsu, China.
J Ethnopharmacol. 2024 Jan 10;318(Pt A):116898. doi: 10.1016/j.jep.2023.116898. Epub 2023 Jul 17.
Cerebral ischemia-reperfusion injury (CIRI) is a complex pathophysiological process involving multiple factors, and becomes the footstone of rehabilitation after ischemic stroke. Sanpian decoction (SPD) has exhibited protective effects against CIRI, migraine, and other cerebral vascular diseases. However, the underlying mechanisms have not been completely elucidated.
This study sought to explore the potential mechanisms underlying the effect of SPD against CIRI.
High-performance liquid chromatography (HPLC) and ultra-high-performance liquid chromatography (UPLC) were carried out to determine the chemical constituents of SPD. A network pharmacology approach combined with experimental verification was conducted to elucidate SPD's multi-component, multi-target, and multi-pathway mechanisms in CIRI occurrence. The pharmacodynamics of the decoction was evaluated by establishing the rat model of middle cerebral artery occlusion/reperfusion (MCAO/R). In vivo and in vitro experiments were carried out, and the therapeutic effects of SPD were performed using 2,3,5-triphenyltetrazolium chloride (TTC) staining, hematoxylin-eosin (HE) staining, and Nissl staining. We used terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining and flow cytometry to evaluate cortex apoptosis. The quantification of mRNA and corresponding proteins were performed using real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) and Western blot respectively.
Our research showed that pretreatment with SPD improved neurological function and inhibited CIRI. Network pharmacology revealed that the hypoxia-inducible factor-1 (HIF-1) signaling pathway and mitogen-activated protein kinase (MAPK) signaling pathway-mediated apoptosis may be associated with CIRI. In vivo and in vitro experiments, we confirmed that SPD increased cerebral blood flow, improved neural function, and reduced neural apoptosis via up-regulating the expression of sirtuin 1 (SIRT1) and down-regulating phospho-extracellular regulated protein kinases (p-ERK)/ERK and HIF-1α levels in CIRI rats.
Taken together, the present study systematically revealed the potential targets and signaling pathways of SPD in the treatment of CIRI using in silico prediction and verified the therapeutic effects of SPD against CIRI via ameliorating apoptosis by regulating SIRT1/ERK/HIF-1α.
脑缺血再灌注损伤（CIRI）是一个涉及多种因素的复杂病理生理过程，是缺血性中风后康复的基石。三偏瘫复方可对CIRI、偏头痛及其他脑血管疾病发挥保护作用。然而，其潜在机制尚未完全阐明。
本研究旨在探索三偏瘫复方抗CIRI作用的潜在机制。
采用高效液相色谱（HPLC）和超高效液相色谱（UPLC）法测定三偏瘫复方的化学成分。采用网络药理学方法结合实验验证，阐明三偏瘫复方在CIRI发生中的多成分、多靶点、多途径作用机制。通过建立大鼠大脑中动脉闭塞/再灌注（MCAO/R）模型评估该复方汤剂的药效学。进行体内和体外实验，采用2,3,5-三苯基氯化四氮唑（TTC）染色、苏木精-伊红（HE）染色和尼氏染色观察三偏瘫复方的治疗效果。采用末端脱氧核苷酸转移酶dUTP缺口末端标记（TUNEL）染色和流式细胞术评估皮质细胞凋亡情况。分别采用实时定量逆转录聚合酶链反应（RT-qPCR）和蛋白质免疫印迹法对mRNA和相应蛋白进行定量分析。
我们的研究表明，三偏瘫复方预处理可改善神经功能并抑制CIRI。网络药理学研究显示，缺氧诱导因子-1（HIF-1）信号通路和丝裂原活化蛋白激酶（MAPK）信号通路介导的细胞凋亡可能与CIRI有关。体内和体外实验均证实，三偏瘫复方可通过上调沉默调节蛋白1（SIRT1）的表达、下调磷酸化细胞外调节蛋白激酶（p-ERK）/ERK和HIF-1α水平，增加脑血流量、改善神经功能并减少神经细胞凋亡。
综上所述，本研究通过计算机预测系统揭示了三偏瘫复方治疗CIRI的潜在靶点和信号通路，并通过调节SIRT1/ERK/HIF-1α改善细胞凋亡，验证了三偏瘫复方对CIRI的治疗作用。
