Department of Pharmacy, First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, National Clinical Research Center for Chinese Medicine Acupuncture and Moxibustion, Tianjin, China.
National Health Commission's Key Laboratory for Critical Care Medicine, Tianjin First Central Hospital, Tianjin, China.
Biomed Res. 2024;45(5):197-207. doi: 10.2220/biomedres.45.197.
Our study explored the therapeutic effect and the mechanism of quercetin against hypoxia/reoxygenation (H/R)-induced injury in human coronary artery endothelial cells (CAECs). Quercetin was selected as a potential component for the BuShenKangShuaiPian formula (BSKSP) treatment via the Network pharmacology analysis. Cell viability and reactive oxygen species (ROS) production were measured by CCK8 assay and immunofluorescence, respectively. The expression of Bax, Bcl-2, Cle-caspase-3, cytochrome c (Cyt-C), NF-E2-related factor 2 (Nrf2), and heme oxygenase-1 (HO-1) protein was quantified by western blotting. The superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA) activity, mtDNA copy number, and ATP production were measured via corresponding kits. Quercetin was selected from the BSKSP for its high degree value (Degree value: 22). Besides, quercetin protected CAECs against H/R-induced cytotoxicity and apoptosis. The H/R-induced increased ROS level, ATP production, Cyt-C release, and decreased mtDNA copy number were removed by the quercetin. Moreover, quercetin upregulated the Nrf2/ HO-1 axis, SOD, and CAT activity, and downregulated MDA levels in H/R treated CAECs, while knockdown Nrf2 reversed the protection of quercetin against H/R-induced oxidative stress, mitochondrial damage, and apoptosis. Quercetin protects CAECs against H/R-induced mitochondrial apoptosis via the Nrf2/HO-1 axis, which innovatively suggests the therapeutic potential of quercetin for coronary heart disease (CHD) treatment.
我们的研究探讨了槲皮素对人冠状动脉内皮细胞（CAEC）缺氧/复氧（H/R）损伤的治疗作用和机制。通过网络药理学分析，选择槲皮素作为补肾抗衰片（BSKSP）治疗的潜在成分。通过 CCK8 测定法和免疫荧光分别测定细胞活力和活性氧（ROS）的产生。通过 Western blot 定量测定 Bax、Bcl-2、Cle-caspase-3、细胞色素 c（Cyt-C）、核因子 E2 相关因子 2（Nrf2）和血红素加氧酶-1（HO-1）蛋白的表达。通过相应的试剂盒测定超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、丙二醛（MDA）活性、线粒体 DNA 拷贝数和 ATP 产生。选择槲皮素是因为它的程度值（Degree value：22）高。此外，槲皮素可保护 CAEC 免受 H/R 诱导的细胞毒性和细胞凋亡。H/R 诱导的 ROS 水平升高、ATP 产生、Cyt-C 释放和 mtDNA 拷贝数降低被槲皮素消除。此外，槲皮素上调了 Nrf2/HO-1 轴、SOD 和 CAT 活性，降低了 H/R 处理的 CAEC 中的 MDA 水平，而敲低 Nrf2 逆转了槲皮素对 H/R 诱导的氧化应激、线粒体损伤和细胞凋亡的保护作用。槲皮素通过 Nrf2/HO-1 轴保护 CAEC 免受 H/R 诱导的线粒体凋亡，这创新性地提示了槲皮素治疗冠心病（CHD）的潜力。
