Ling Xiaoying, Zhou Jie, Jin Tianzi, Xu Weichen, Sun Xun, Li Weifeng, Ding Yali, Liang Miaomiao, Zhu Chenbi, Zhao Peipei, Hu Chanchan, Yuan Bin, Xie Tong, Tao Jialei
Department of Pediatrics, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, China.
Jiangsu Key Laboratory of Pediatric Respiratory Disease, Institute of Pediatrics, Medical Metabolomics Center, Nanjing University of Chinese Medicine, Nanjing, China.
Front Pharmacol. 2022 Aug 19;13:870928. doi: 10.3389/fphar.2022.870928. eCollection 2022.
Necroptosis and inflammation are closely related to the pathogenesis of respiratory syncytial virus (RSV). Acteoside (AC), a natural phenylpropanoid glycoside from Kuding Tea, has significant anti-RSV effect. However, the roles of AC on RSV-induced lung necroptosis and inflammation are yet to be elucidated. The effects of AC were investigated in BALB/c mice and A549 cells. Lung histopathology was observed through H&E staining. The viral titer was assessed via plaque assay. The RSV-F expression was determined by RT-qPCR and immunohistochemistry assay. The levels of cytokines were detected by ELISA and RT-qPCR. The necroptosis rate and mitochondrial membrane potential were evaluated via flow cytometry. The expressions of HMGB1/NF-κB and RIP1/RIP3/MLKL/PGAM5/DRP1 were detected by western blot. Additionally, untargeted metabolomics was conducted to investigate the metabolic profiles and related metabolic pathways via Gas Chromatography-Mass Spectrometry. The results showed that compared with the RSV-infected group, AC treatment significantly attenuated lung pathological damage, virus replication, and cytokines levels. AC also alleviated RSV-induced necroptosis and mitochondrial dysfunction . Moreover, AC treatment down-regulated the expression of HMGB1, p-Iκbα/Iκbα, p-p65/p65, RIP1, RIP3, MLKL, PGAM5, and DRP1. Furthermore, metabolomic analyses suggested that the perturbations in major metabolites of AC therapy were related to variations in amino acid and energy metabolism. Our findings validated the beneficial effects of AC in suppressing necroptosis and regulating metabolism, suggesting AC may be a new drug candidate for RSV infection.
坏死性凋亡与炎症与呼吸道合胞病毒（RSV）的发病机制密切相关。毛蕊花糖苷（AC）是一种从苦丁茶中提取的天然苯丙素苷，具有显著的抗RSV作用。然而，AC对RSV诱导的肺坏死性凋亡和炎症的作用尚待阐明。本研究在BALB/c小鼠和A549细胞中研究了AC的作用。通过苏木精-伊红（H&E）染色观察肺组织病理学变化。通过空斑试验评估病毒滴度。通过逆转录定量聚合酶链反应（RT-qPCR）和免疫组织化学测定法测定RSV-F表达。通过酶联免疫吸附测定（ELISA）和RT-qPCR检测细胞因子水平。通过流式细胞术评估坏死性凋亡率和线粒体膜电位。通过蛋白质免疫印迹法检测高迁移率族蛋白B1（HMGB1）/核因子κB（NF-κB）以及受体相互作用蛋白1（RIP1）/RIP3/混合谱系激酶结构域样蛋白（MLKL）/磷酸甘油酸变位酶5（PGAM5）/动力相关蛋白1（DRP1）的表达。此外，采用非靶向代谢组学通过气相色谱-质谱联用技术研究代谢谱和相关代谢途径。结果表明，与RSV感染组相比，AC治疗显著减轻了肺组织病理损伤、病毒复制和细胞因子水平。AC还减轻了RSV诱导的坏死性凋亡和线粒体功能障碍。此外，AC治疗下调了HMGB1、磷酸化IκBα/IκBα、磷酸化p65/p65、RIP1、RIP3、MLKL、PGAM5和DRP1的表达。此外，代谢组学分析表明，AC治疗中主要代谢物的扰动与氨基酸和能量代谢的变化有关。我们的研究结果证实了AC在抑制坏死性凋亡和调节代谢方面的有益作用，表明AC可能是一种用于RSV感染的新型候选药物。
