Oxymatrine Inhibits Homocysteine-Mediated Autophagy via MIF/mTOR Signaling in Human Umbilical Vein Endothelial Cells
Oxymatrine Inhibits Homocysteine-Mediated Autophagy via MIF/mTOR Signaling in Human Umbilical Vein Endothelial Cells
复制标题
氧化苦参碱通过人脐静脉内皮细胞中的 MIF/mTOR 信号抑制同型半胱氨酸介导的自噬
DOI：
10.1159/000487912
复制
发表时间：
2018-01-01
影响因子：
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通讯作者：
Yi, Minhan
中科院分区：
文献类型：
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作者：
Zhang, Yanyan;Zhang, Yuan;Yi, Minhan
背景/目的：同型半胱氨酸（Hcy）代谢的遗传或营养缺陷导致Hcy及其代谢产物在血液中蓄积。这可能导致高同型半胱氨酸血症（HHcy），这是心血管疾病的独立危险因素。研究表明，高同型半胱氨酸导致内皮功能障碍，动脉粥样硬化的标志，这可能解释这种联系。确切的机制尚不清楚，但很有可能是过度的HHCy诱导的自噬。自噬在心肌缺血/再灌注损伤中的作用已得到较好的研究，氧化苦参碱（Oxymatrine，OMT）是一种喹嗪啶生物碱，可通过抑制自噬来保护心肌细胞免受I/R损伤。本研究的目的是确定OMT是否抑制HHcy中的自噬。研究方法：通过透射电镜观察Hcy处理的HUVEC细胞在存在和不存在OMT的情况下的自噬，并通过western blotting和qRT-PCR确定自噬程度。使用小干扰RNA（siRNA）来测定巨噬细胞移动抑制因子（MIF）抑制的效率。Western blotting和流式细胞仪检测细胞凋亡。结果：OMT抑制Hcy暴露过程中HUVECs的自噬、MIF和mTOR，这取决于剂量。siRNA介导的MIF敲低降低了Hcy诱导的自噬，而3-甲基腺苷和雷帕霉素的施用表明它们也诱导自噬。此外，OMT剂量依赖性地抑制Hcy诱导的HUVEC凋亡/死亡。结论：这些结果表明，Hcy可通过MIF/mTOR信号通路诱导自噬激活的HUVEC凋亡/死亡，OMT可逆转该信号通路。我们的研究结果提供了一个新的见解OMT在预防Hcy诱导的HUVEC损伤和死亡的功能作用。
Background/Aims: Genetic or nutritional deficiencies in homocysteine (Hcy)metabolism lead to the accumulation of Hcy and its metabolites in the blood. This can lead to hyperhomocysteinemia (HHcy), which is an independent risk factor for cardiovascular disease. Studies have shown that HHcy leads to endothelial dysfunction, a hallmark of atherosclerosis, which may explain this link. The precise mechanism remains unclear, but a strong possibility is excessive HHCy-induced autophagy. Autophagy has been better studied in ischemia/reperfusion (I/R) injuries, and previous work showed that Oxymatrine (OMT), a quinolizidine alkaloid, protects cells against myocardial I/R injury by inhibiting autophagy. The aim of this study was to determine whether OMT inhibits autophagy in HHcy. Methods: Autophagy in HUVEC cells treated with Hcy in the presence and absence of OMT was visualized bytransmission electron microscopy and the degree was determined by western blotting and qRT-PCR. Small interfering RNA (siRNA)was used to determine the efficiency of Macrophage migration inhibitory factor (MIF) inhibition. Cell apoptosis wasdetected by western blotting and flow cytometric analysis. Results: OMT inhibited autophagy, MIF, and mTOR in HUVECs during Hcy exposure, depending on the dose. siRNA-mediated MIF knockdown decreased Hcy-induced autophagy, while administration of 3-methyladenosine and rapamycin showed that they also induce autophagy. Furthermore, OMT dose-dependently inhibited the Hcy-induced HUVEC apoptosis/death. Conclusions: These results suggest that Hcy can evokeautophagy-activated HUVEC apoptosis/death via a MIF/mTOR signaling pathway, which can be reversed by OMT. Our results provide a new insight into a functional role of OMT in the prevention of Hcy-induced HUVEC injury and death.