Department of Comprehensive health care, Affiliated Hangzhou First People's Hospital, Zhejiang University School of Medicine.
PICC P&C (PICC Property and Casualty Company Limited) Zhejiang Branch, Hangzhou, China.
Neuroreport. 2022 Nov 2;33(16):714-722. doi: 10.1097/WNR.0000000000001837. Epub 2022 Sep 19.
Amyloid β-protein (Aβ)-induced apoptosis and oxidative stress of human brain microvascular endothelial cells(BMECs) are contributors to the development of Alzheimer's disease (AD). Acteoside has shown its therapeutic potential for AD treatment. Therefore, this study investigated the effect of acteoside on Aβ-induced blood-brain barrier damage, oxidative stress and apoptosis as well as to explore the underlying mechanisms through network pharmacology.
The study used Aβ to induce human BMECs to construct an in-vitro injury model. Following treatment with acteoside, transendothelial electrical resistance (TEER), RT-qPCR and Western blot were used to evaluate the permeability of BMECs. The apoptosis level was detected by TUNEL and Western blot, ROS assay kit was used for the detection of reactive oxygen species (ROS) expression. The let-7g-5p expression level was detected by RT-qPCR. After additional treatment with let-7g-5p inhibitor, corresponding assays were performed again. Finally, network pharmacology was used to verify the mechanism.
Acteoside decreased the permeability, oxidative stress and cell apoptosis of Aβ-stimulated cells. More importantly, acteoside-activated let-7g-5p and additional treatment with let-7g-5p inhibitor abated the effects of acteoside on Aβ-induced permeability, oxidative stress and apoptosis of Aβ-stimulated BMECs. According to network pharmacology, 233 targeted genes of acteoside and 122 potential targets of let-7g-5p were determined by screening several databases, and two targets called Casp-3 and ITGB3 were obtained after taking the intersection.
In conclusion, these results reveal that acteoside-activated let-7g-5p attenuating Aβ-induced increased permeability and apoptosis of human BMECs.
淀粉样蛋白β（Aβ）诱导的人脑血管内皮细胞（BMECs）凋亡和氧化应激是阿尔茨海默病（AD）发病的原因之一。獐牙菜苦苷已显示出治疗 AD 的潜力。因此，本研究通过网络药理学探讨了獐牙菜苦苷对 Aβ诱导的血脑屏障损伤、氧化应激和凋亡的影响及其作用机制。
采用 Aβ诱导人 BMECs 构建体外损伤模型，用獐牙菜苦苷处理后，通过跨内皮电阻（TEER）、实时定量 PCR（RT-qPCR）和 Western blot 检测 BMECs 的通透性。TUNEL 和 Western blot 检测细胞凋亡水平，活性氧（ROS）检测试剂盒检测 ROS 表达。RT-qPCR 检测 let-7g-5p 表达水平。用 let-7g-5p 抑制剂处理后，再次进行相应检测。最后，用网络药理学验证机制。
獐牙菜苦苷降低了 Aβ刺激细胞的通透性、氧化应激和细胞凋亡。更重要的是，獐牙菜苦苷激活了 let-7g-5p，并用 let-7g-5p 抑制剂处理后减弱了獐牙菜苦苷对 Aβ诱导的 BMECs 通透性、氧化应激和凋亡的作用。通过筛选几个数据库，网络药理学确定了獐牙菜苦苷的 233 个靶基因和 let-7g-5p 的 122 个潜在靶标，经过取交集后得到了两个名为 Casp-3 和 ITGB3 的靶标。
总之，这些结果表明，獐牙菜苦苷激活 let-7g-5p 可减轻 Aβ诱导的人 BMECs 通透性增加和凋亡。
