The Artemisinin Derivative Artesunate Inhibits Corneal Neovascularization by Inducing ROS-Dependent Apoptosis in Vascular Endothelial Cells
The Artemisinin Derivative Artesunate Inhibits Corneal Neovascularization by Inducing ROS-Dependent Apoptosis in Vascular Endothelial Cells
复制标题
青蒿素衍生物青蒿琥酯通过诱导血管内皮细胞中ROS依赖性细胞凋亡来抑制角膜新生血管形成。
DOI：
10.1167/iovs.12-11068
复制
发表时间：
2013-05-01
影响因子：
4.4
通讯作者：
Gao, Guoquan
中科院分区：
文献类型：
--
作者：
Cheng, Rui;Li, Cen;Gao, Guoquan
目的。无需治疗干预，角膜新血管形成迅速损害了视力，并且是失明的主要原因。据报道，尚未研究青霉素会抑制肿瘤中的血管生成，尽管尚未研究青霉素对非肿瘤血管生成的作用。这项研究旨在研究临床酸磷藻对角膜新血管形成的影响，并描述其基本的作用机理。用碱角膜烧伤的大鼠用赤臂治疗11天。通过测量大鼠角膜血管的长度和面积来评估角膜新血管形成。将凋亡细胞用膜剂和碘化丙氨酸（PI）染色，并通过流式细胞仪分析进行测量。通过Western blot Analysis.ults评估了与凋亡相关和p38促丝分裂原激活蛋白激酶（p38mapk）信号传导。血氧酸盐通过特异性诱导血管内皮细胞的凋亡来显着抑制角膜新血管形成和炎症。在血管内皮细胞中，敏捷酸盐提高了BAX/BCL-2的比率，线粒体膜电位降低，细胞色素C的刺激释放以及caspase 9和3的裂解，这表明涉及线粒体的凋亡途径。血氧酸盐激活的p38mapk和特定的p38MAPK抑制剂抑制了固醇诱导的内皮细胞凋亡。活性氧（ROS）水平通过丙酸苯酚升高。 N-乙酰基L-半胱氨酸阻塞P38MAPK激活，并保护内皮细胞免受炎症诱导的细胞凋亡。亚铁盐提高了ROS水平，并升高了氯吡酯对内皮细胞的细胞毒性作用，而铁螯合剂脱铁胺降低了ROS水平和氯酸锰酸盐诱导的凋亡。青蒿酯对FAS，FAS配体或caspase 8裂解的表达没有影响。这些结果表明，白蚁酸会通过铁/ROS依赖性p38MAPK-MITOCHRIAL途径诱导内皮细胞的凋亡。
PURPOSE
Without therapeutic intervention, corneal neovascularization rapidly compromises visual acuity, and is a leading cause of blindness. Artesunate was reported to inhibit angiogenesis in tumors, although, the effects of artesunate on nontumor angiogenesis have not been investigated. This study was designed to investigate the effect of artesunate on corneal neovascularization and delineate its underlying mechanism of action.
METHODS
Rats with alkali-burned corneas were treated with artesunate for 11 days. Corneal neovascularization was evaluated by measuring the length and area of corneal vasculature in the rats. Apoptotic cells were stained with AnnexinV and propidine iodide (PI), and measured with flow cytometry analysis. Apoptosis-related and p38 mitogen-activated protein kinases (p38MAPK) signaling were evaluated by Western blot analysis.
RESULTS
Artesunate significantly inhibited corneal neovascularization and inflammation via specifically inducing apoptosis of vascular endothelial cells. In vascular endothelial cells, artesunate increased the Bax/Bcl-2 ratio, reduced mitochondrial membrane potential, stimulated release of cytochrome C, and cleavage of caspase 9 and 3, suggesting that the mitochondrial apoptotic pathway was involved. Artesunate activated p38MAPK, and specific p38MAPK inhibitors suppressed artesunate-induced apoptosis in endothelial cells. Reactive oxygen species (ROS) levels were increased by artesunate. N-acetyl-L-cysteine blocked p38MAPK activation and protected endothelial cells from artesunate-induced apoptosis. Ferrous salt increased ROS levels and elevated the cytotoxic effect of artesunate on endothelial cells, while the iron chelating agent deferoxamine decreased ROS levels and artesunate-induced apoptosis. Artesunate had no effect on expression of Fas, Fas Ligand, or caspase 8 cleavage.
CONCLUSIONS
These results suggest that artesunate induces apoptosis of endothelial cells via an iron/ROS-dependent p38MAPK-mitochondrial pathway.