Luo Caiying, Zhang Li, Wang Aiyun, Wang Yuting, Cai Jing, Ni Ruoyan, Yang Renhua, He Sijin, Gu Lihua, Chen Peng
School of Pharmaceutical Sciences and Yunnan Key Laboratory of Pharmacology for Natural Products, Kunming Medical University Kunming 650500, Yunnan, China.
College of Modern Biomedical Industry, Kunming Medical University Kunming 650500, Yunnan, China.
Am J Transl Res. 2025 Jul 15;17(7):4894-4911. doi: 10.62347/MLZI7617. eCollection 2025.
This study aimed to investigate the effects of Shu-Tiao Qi-Ji Decoction (STQJD) on a rat model of cerebral ischemia-reperfusion injury and to elucidate its mechanism of action.
The therapeutic effects of STQJD on ischemic stroke (IS) were assessed using neurobehavioral scores, 2,3,5-Triphenyltetrazolium chloride (TTC), Nissl and Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining. Network pharmacology was used to investigate the potential mechanisms of STQJD. Reactive Oxygen Species (ROS) content and expression levels of inflammatory markers in rat serum were detected. The mRNA and protein expression levels of Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), NADPH oxidase 4 (NOX4), Tumor necrosis factor-alpha (TNF-α), NOD-like receptor family pyrin domain containing 3 (NLRP3), Apoptosis-associated speck-like protein containing a CARD (ASC), Toll-like receptor 4 (TLR4), and Cysteine-dependent aspartate-specific protease-1 (Caspase-1) in the brain tissue of rats were detected using RT-qPCR and Western blotting. Immunofluorescence was used to detect NF-κB and TNF-α protein expression.
STQJD markedly improved neurological function, reduced cerebral infarct volume, and mitigated neuronal damage and apoptosis in rats subjected to Middle Cerebral Artery Occlusion (MCAO). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses revealed that STQJD's anti-IS effects are linked to NOX4/TLR4/NF-κB/TNF-α signaling pathways. STQJD reduced serum levels of inflammatory markers, including TNF-α, Interleukin-1 beta (IL-1β), and Interleukin-18 (IL-18), as well as mRNA and protein expression of TLR4, NOX4, NLRP3, NF-κB, ASC, TNF-α, and Caspase-1 in the brain tissues of MCAO rat levels.
STQJD effectively ameliorates IS in MCAO/R rats by modulating the NOX4/TLR4/NF-κB/TNF-α signaling pathway, reducing inflammation and oxidative damage, and exerting neuroprotective effects.
本研究旨在探讨舒调气机汤（STQJD）对大鼠脑缺血再灌注损伤模型的影响，并阐明其作用机制。
采用神经行为学评分、2,3,5-氯化三苯基四氮唑（TTC）、尼氏染色和末端脱氧核苷酸转移酶介导的dUTP缺口末端标记（TUNEL）染色评估STQJD对缺血性中风（IS）的治疗效果。运用网络药理学研究STQJD的潜在作用机制。检测大鼠血清中活性氧（ROS）含量及炎症标志物的表达水平。采用逆转录定量聚合酶链反应（RT-qPCR）和蛋白质免疫印迹法检测大鼠脑组织中活化B细胞核因子κB（NF-κB）、烟酰胺腺嘌呤二核苷酸磷酸氧化酶4（NOX4）、肿瘤坏死因子-α（TNF-α）、含NOD样受体家族吡咯结构域蛋白3（NLRP3）、含半胱天冬酶激活和招募结构域的凋亡相关斑点样蛋白（ASC）、Toll样受体4（TLR4）和半胱氨酸天冬氨酸特异性蛋白酶-1（Caspase-1）的mRNA和蛋白表达水平。采用免疫荧光法检测NF-κB和TNF-α蛋白表达。
STQJD显著改善大脑中动脉闭塞（MCAO）大鼠的神经功能，减少脑梗死体积，减轻神经元损伤和凋亡。基因本体论（GO）和京都基因与基因组百科全书（KEGG）分析显示，STQJD的抗IS作用与NOX4/TLR4/NF-κB/TNF-α信号通路有关。STQJD降低了血清中炎症标志物的水平，包括TNF-α、白细胞介素-1β（IL-1β）和白细胞介素-18（IL-18），以及MCAO大鼠脑组织中TLR4、NOX4、NLRP3、NF-κB、ASC、TNF-α和Caspase-1的mRNA和蛋白表达水平。
STQJD通过调节NOX4/TLR4/NF-κB/TNF-α信号通路，减轻炎症反应和氧化损伤，发挥神经保护作用，有效改善MCAO/R大鼠的IS。
