Zhu Meixia, Li Jiankuan, Wang Ke, Hao Xuliang, Ge Rui, Li Qingshan
Department of Pharmacy Shanxi, College of Traditional Chinese Medicine, No. 121 University Avenue, Taiyuan 030619, China.
Shanxi Province Academy of Traditional Chinese Medicine, No. 46 Bingzhou West Road, Taiyuan 030012, China.
Molecules. 2016 Mar 21;21(3):356. doi: 10.3390/molecules21030356.
Oxidative stress plays a critical role in endothelial injury and the pathogenesis of diverse cardiovascular diseases, including atherosclerosis. Isoquercitrin (quercetin-3-glucoside), a flavonoid distributed widely in plants, exhibits many biological activities, including anti-allergic, anti-viral, anti-inflammatory, and anti-oxidative effects. In the present study, the inhibitory effect of isoquercitrin on H2O2-induced apoptosis of EA.hy926 cells was evaluated. MTT assays showed that isoquercitrin significantly inhibited H2O2-induced loss of viability in EA.hy926 cells. Hoechst33342/PI and Annexin V-FITC/PI fluorescent double staining indicated that isoquercitrin inhibited H2O2-induced apoptosis of EA.hy926 cells. Western blotting demonstrated that isoquercitrin prevented H2O2-induced increases in cleaved caspase-9 and cleaved caspase-3 expression, while increasing expression of anti-apoptotic protein Mcl-1. Additionally, isoquercitrin significantly increased the expression of p-Akt and p-GSK3β in a dose-dependent manner in EA.hy926 cells. LY294002, a PI3K/Akt inhibitor, inhibited isoquercitrin-induced GSK3β phosphorylation and increase of Mcl-1 expression, which indicated that regulation of isoquercitrin on Mcl-1 expression was likely related to the modulation of Akt activation. These results demonstrated that the anti-apoptotic effect of isoquercitrin on H2O2-induced EA.hy926 cells was likely associated with the regulation of isoquercitrin on Akt/GSK3β signaling pathway and that isoquercitrin could be used clinically to interfere with the progression of endothelial injury-associated cardiovascular disease.
氧化应激在血管内皮损伤及包括动脉粥样硬化在内的多种心血管疾病的发病机制中起关键作用。异槲皮苷（槲皮素 - 3 - 葡萄糖苷）是一种广泛分布于植物中的黄酮类化合物，具有多种生物学活性，包括抗过敏、抗病毒、抗炎和抗氧化作用。在本研究中，评估了异槲皮苷对H2O2诱导的EA.hy926细胞凋亡的抑制作用。MTT分析表明，异槲皮苷显著抑制H2O2诱导的EA.hy926细胞活力丧失。Hoechst33342/PI和Annexin V - FITC/PI荧光双染色表明，异槲皮苷抑制H2O2诱导的EA.hy926细胞凋亡。蛋白质免疫印迹法显示，异槲皮苷可阻止H2O2诱导的裂解型半胱天冬酶 - 9和裂解型半胱天冬酶 - 3表达增加，同时增加抗凋亡蛋白Mcl - 1的表达。此外，异槲皮苷在EA.hy926细胞中以剂量依赖性方式显著增加p - Akt和p - GSK3β的表达。PI3K/Akt抑制剂LY294002抑制了异槲皮苷诱导的GSK3β磷酸化和Mcl - 1表达增加，这表明异槲皮苷对Mcl - 1表达的调节可能与Akt激活的调节有关。这些结果表明，异槲皮苷对H2O2诱导的EA.hy926细胞的抗凋亡作用可能与异槲皮苷对Akt/GSK3β信号通路的调节有关，并且异槲皮苷可在临床上用于干预与血管内皮损伤相关的心血管疾病的进展。
