Department of neurology, Changshu Hospital affiliated to Nanjing University of Chinese Medicine, 6 Huanghe Road, Changshu, Jiangsu 215500, China; Nanjing University of Chinese Medicine, Nanjing 210023, China.
Nanjing University of Chinese Medicine, Nanjing 210023, China.
Biomed Pharmacother. 2023 Dec;168:115598. doi: 10.1016/j.biopha.2023.115598. Epub 2023 Oct 9.
Cerebral infarction (CI) has become one of the leading causes of death and acquired disability worldwide. Astragaloside IV (AST IV), one of the basic components of Astragalus membranaceus, has a protective effect on CI. However, the underlying mechanism has not been conclusively elucidated. Therefore, this study aims to explore the underlying mechanism of AST IV improving brain injury after CI. Middle cerebral artery occlusion (MCAO) and oxygen-glucose deprivation/reoxygenation (OGD/R) were used to simulate cerebral infarction injury in SD rats and HUVECs cells. Neurologic score, Evans blue, TTC and HE staining were used to observe brain injury in rats. Cell viability and migration were measured in vitro. Angiogenesis was detected by immunofluorescence and tube formation assay, and cell cycle was detected by flow cytometry. Western blot was used to find the expression of related proteins. Molecular docking, virtual mutation, site-directed mutagenesis, MST, and lentivirus silencing were used for target validation. The results showed that AST IV alleviated neurological impairment and promoted angiogenesis after CI. Moreover, AST IV greatly increased the transcription levels of SIRT6 and SIRT7, but had no effect on SIRT1-SIRT5, and promoted cell viability, migration, angiogenesis and S phase ratio in OGD/R-induced HUVECs. Furthermore, AST IV up-regulated the protein expressions of CDK4, cyclin D1, VEGFA and VEGF2R. Interestingly, AST IV not only bound to SIRT7, but also increased the expression of SIRT7. Silencing SIRT7 by lentivirus neutralizes the positive effects of AST IV. Taken together, the present study revealed that AST IV may improve brain tissue damage after CI by targeting SIRT7/VEGFA signaling pathway to promote angiogenesis.
脑梗死(CI)已成为全球范围内导致死亡和获得性残疾的主要原因之一。黄芪甲苷(AST IV)是黄芪的基本成分之一,对 CI 具有保护作用。然而,其潜在机制尚未得到明确阐明。因此,本研究旨在探讨 AST IV 改善 CI 后脑损伤的潜在机制。采用大脑中动脉闭塞(MCAO)和氧葡萄糖剥夺/再氧合(OGD/R)模拟 SD 大鼠和 HUVECs 细胞的脑梗死损伤。神经功能评分、伊文思蓝、TTC 和 HE 染色用于观察大鼠脑损伤。体外测量细胞活力和迁移。免疫荧光和管形成试验检测血管生成,流式细胞术检测细胞周期。Western blot 用于检测相关蛋白的表达。分子对接、虚拟突变、定点突变、MST 和慢病毒沉默用于验证靶点。结果表明,AST IV 减轻了 CI 后的神经功能损伤并促进了血管生成。此外,AST IV 极大地增加了 SIRT6 和 SIRT7 的转录水平,但对 SIRT1-SIRT5 没有影响,并促进了 OGD/R 诱导的 HUVECs 中的细胞活力、迁移、血管生成和 S 期比例。此外,AST IV 上调了 CDK4、cyclin D1、VEGFA 和 VEGF2R 的蛋白表达。有趣的是,AST IV 不仅与 SIRT7 结合,还增加了 SIRT7 的表达。慢病毒沉默 SIRT7 中和了 AST IV 的阳性作用。总之,本研究表明,AST IV 可能通过靶向 SIRT7/VEGFA 信号通路促进血管生成来改善 CI 后的脑组织损伤。