Background: 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.
Methods: 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.
Results: 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.
Conclusion: FAME possesses significant anti-inflammatory and anti-oxidative stress activities and alleviates CIRI through the PI3K/HIF-1α/VEGF signaling pathway.



中文翻译：

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阿魏酸甲酯通过调节 PI3K/HIF-1α/VEGF 信号通路减轻大鼠脑缺血再灌注损伤


背景：脑缺血再灌注损伤（CIRI）可能恶化脑组织的炎症反应和氧化应激。根据既往研究，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，具有显着的抗氧化应激和神经保护功能，可有效缓解CIRI，但其作用机制尚不清楚。

方法：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MACO模型，通过脑组织病理学染色和脑梗死面积评估FAME治疗CIRI的治疗效果。第三，利用RNA-Seq和网络药理学在分子水平预测FAME可能的靶点和机制。最后，通过分子对接可视化、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。

结果：体外实验结果表明，FAME可显着降低细胞内TNF-α、IL-1β和ROS的含量，抑制细胞内COX-2和iNOS蛋白表达（p< 0.01）。 FAME 被证明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大鼠脑梗死面积减少，Bcl-2和VEGF含量增加(p<<0.05)。网络药理学和RNA-Seq表明FAME减轻CIRI可能是通过PI3K-AKT和HIF-1信号通路。通过分子对接和Western Blotting验证FAME组大鼠脑组织中HIF-1α、VEGF、p-PI3K、p-AKT蛋白表达增强。

结论： FAME具有显着的抗炎和抗氧化应激活性，并通过PI3K/HIF-1α/VEGF信号通路缓解CIRI。