Wang Lele, Liu Canwen, Wang Linlin, Tang Biao
Basic Research Center of Integrated Chinese & Western Medicine for Prevention & Treatment of Vascular Diseases, Hunan University of Chinese Medicine, Changsha, Hunan, 410208, China.
Eur J Pharmacol. 2023 Apr 5;944:175516. doi: 10.1016/j.ejphar.2023.175516. Epub 2023 Feb 7.
Cerebral ischaemia-reperfusion injury (CIRI) is a critical component of ischaemic stroke pathogenesis. Ferroptosis contributes to and aggravates CIRI, whereas the P62/Kelch-like ECH-associated protein 1 (Keap1)/NF-E2-related factor 2 (Nrf2) pathway exerts neuroprotective effects. Astragaloside IV (AST IV) is the primary active ingredient of Astragalus, an herb with anti-CIRI properties used in traditional Chinese medicine. However, the mechanism of its anti-CIRI action is unclear. This study examined the mechanisms underlying the anti-CIRI action of AST IV using a combination of in vitro and in vivo approaches. We established an erastin-induced ferroptosis model, oxygen and glucose deprivation/reoxygenation (OGD/R)-induced model in SH-SY5Y cells, and middle cerebral artery occlusion-reperfusion (MCAO/R) model using Sprague-Dawley rats. The extent of cell damage and brain damage in rats, ferroptosis indicator changes, and expression of P62, Keap1, and Nrf2 were investigated. AST IV inhibited erastin-induced ferroptosis, attenuated OGD/R-induced cell damage, and ameliorated sensorimotor dysfunction and injury in the MCAO/R model. Further, AST IV promoted Nrf2 activation, inhibited ferroptosis, and reduced cell damage. Notably, these effects were inhibited by ML385, an Nrf2 inhibitor. AST IV increased the P62 and Nrf2 levels and decreased the Keap1 levels. P62 silencing reduced the effects of AST IV on the P62/Keap1/Nrf2 pathway and ferroptosis. Our findings suggest that AST IV mitigates CIRI by inhibiting ferroptosis via activation of the P62/Keap1/Nrf2 pathway. This study provides an important scientific basis and direction for the application and research of AST IV and provides new potential targets and ideas for the study of the pathological mechanism of CIRI.
脑缺血再灌注损伤（CIRI）是缺血性脑卒中发病机制的关键组成部分。铁死亡促进并加重CIRI，而P62/ Kelch样ECH相关蛋白1（Keap1）/核因子E2相关因子2（Nrf2）通路发挥神经保护作用。黄芪甲苷（AST IV）是黄芪的主要活性成分，黄芪是一种用于传统中药中具有抗CIRI特性的草药。然而，其抗CIRI作用的机制尚不清楚。本研究采用体外和体内相结合的方法，研究了AST IV抗CIRI作用的机制。我们建立了erastin诱导的铁死亡模型、氧糖剥夺/复氧（OGD/R）诱导的SH-SY5Y细胞模型以及使用Sprague-Dawley大鼠的大脑中动脉闭塞/再灌注（MCAO/R）模型。研究了大鼠的细胞损伤和脑损伤程度、铁死亡指标变化以及P62、Keap1和Nrf2的表达。AST IV抑制erastin诱导的铁死亡，减轻OGD/R诱导的细胞损伤，并改善MCAO/R模型中的感觉运动功能障碍和损伤程度。此外，AST IV促进Nrf2激活，抑制铁死亡，并减少细胞损伤。值得注意的是，这些作用被Nrf2抑制剂ML385所抑制。AST IV增加了P62和Nrf2水平，降低了Keap1水平。P62基因沉默降低了AST IV对P62/Keap1/Nrf2通路和铁死亡的影响。我们的研究结果表明，AST IV通过激活P62/Keap1/Nrf2通路抑制铁死亡来减轻CIRI。本研究为AST IV的应用和研究提供了重要的科学依据和方向，并为CIRI病理机制的研究提供了新的潜在靶点和思路。
