Zhou Peijie, Yu Shangshang, Wang Xuan, Zhang Xiaofei, Guo Dongyan, Zhao Chongbo, Cheng Jiangxue, Wang Jing, Sun Jing
Department of Pharmaceutics, College of Pharmacy, The Key Laboratory of Basic and New Drug Research of Traditional Chinese Medicine, Shaanxi Provincial Engineering Technology Research Center for Traditional Chinese Medicine Decoction Pieces, Shaanxi University of Chinese Medicine, Xianyang, Shaanxi, People's Republic of China.
J Inflamm Res. 2024 Aug 29;17:5741-5762. doi: 10.2147/JIR.S473665. eCollection 2024.
Cerebral ischaemia-reperfusion injury (CIRI) could worsen the inflammatory response and oxidative stress in brain tissue. According to previous studies, ferulic acid methyl ester (FAME), as the extract with the strongest comprehensive activity in the traditional Chinese medicine Huang Hua oil dot herb, has significant anti-oxidative stress and neuroprotective functions, and can effectively alleviate CIRI, but its mechanism of action is still unclear.
Firstly, the pharmacological effects of FAME were investigated by in vitro oxidative stress and inflammatory experiments. Secondly, evaluate the therapeutic effects of FAME in the treatment of CIRI by brain histopathological staining and cerebral infarct area by replicating the in vivo MACO model. Thirdly, RNA-Seq and network pharmacology were utilized to predict the possible targets and mechanisms of FAME for CIRI at the molecular level. Finally, the expression of key target proteins, as well as the key regulatory relationships were verified by molecular docking visualization, Western Blotting and immunohistochemistry.
The results of in vitro experiments concluded that FAME could significantly reduce the content of TNF-α, IL-1β and ROS, inhibiting COX-2 and iNOS protein expression in cells(p<0.01). FAME was demonstrated to have anti-oxidative stress and anti-inflammatory effects. The results of in vivo experiments showed that after the administration of FAME, the area of cerebral infarction in rats with CIRI was reduced, the content of Bcl-2 and VEGF was increased(p<0.05). Network pharmacology and RNA-Seq showed that the alleviation of CIRI by FAME may be through PI3K-AKT and HIF-1 signaling pathway. Enhanced expression of HIF-1α, VEGF, p-PI3K, p-AKT proteins in the brain tissues of rats in the FAME group was verified by molecular docking and Western Blotting.
FAME possesses significant anti-inflammatory and anti-oxidative stress activities and alleviates CIRI through the PI3K/HIF-1α/VEGF signaling pathway.
脑缺血再灌注损伤（CIRI）可加重脑组织的炎症反应和氧化应激。根据以往研究，阿魏酸甲酯（FAME）作为中药黄花油点草中综合活性最强的提取物，具有显著的抗氧化应激和神经保护作用，能有效减轻CIRI，但其作用机制尚不清楚。
首先，通过体外氧化应激和炎症实验研究FAME的药理作用。其次，通过复制体内大脑中动脉闭塞（MACO）模型，采用脑组织病理染色和脑梗死面积评估FAME治疗CIRI的疗效。第三，利用RNA测序（RNA-Seq）和网络药理学在分子水平预测FAME治疗CIRI的潜在靶点和机制。最后，通过分子对接可视化、蛋白质免疫印迹法（Western Blotting）和免疫组化验证关键靶蛋白的表达以及关键调控关系。
体外实验结果表明，FAME可显著降低肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）和活性氧（ROS）含量，抑制细胞中环氧合酶-2（COX-2）和诱导型一氧化氮合酶（iNOS）蛋白表达（p<0.01）。证明FAME具有抗氧化应激和抗炎作用。体内实验结果显示，给予FAME后，CIRI大鼠的脑梗死面积减小，Bcl-2和血管内皮生长因子（VEGF）含量增加（p<0.05）。网络药理学和RNA-Seq显示，FAME减轻CIRI可能是通过磷脂酰肌醇-3激酶-蛋白激酶B（PI3K-AKT）和低氧诱导因子-1（HIF-1）信号通路。通过分子对接和Western Blotting验证了FAME组大鼠脑组织中HIF-1α、VEGF、磷酸化PI3K（p-PI3K）、磷酸化AKT（p-AKT）蛋白表达增强。
FAME具有显著的抗炎和抗氧化应激活性，并通过PI3K/HIF-1α/VEGF信号通路减轻CIRI。
