Anagliptin prevents apoptosis of human umbilical vein endothelial cells by modulating NOX-4 signaling pathways
Anagliptin prevents apoptosis of human umbilical vein endothelial cells by modulating NOX-4 signaling pathways
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阿格列汀通过调节NOX-4信号通路预防人脐静脉内皮细胞凋亡
DOI：
10.1016/j.biopha.2018.04.187
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发表时间：
2018
影响因子：
7.5
通讯作者：
Chen Chang
中科院分区：
文献类型：
--
作者：
Li Qi;Li Jiade;Liu Yanli;Zhang Mingyu;Chen Chang
二肽基肽酶IV(DPP-IV)抑制剂是一种新型的口服降糖药物。本研究旨在探讨Anagliptin对人脐静脉内皮细胞(HUVECs)的抗凋亡作用。培养的人脐静脉内皮细胞与Anagliptin预先孵育，然后用过氧化氢(H_2O_2)诱导细胞凋亡。用活细胞、活/死染色和流式细胞仪检测细胞的凋亡率。将含人NADPH氧化酶(NOX)4的真核表达载体或空载体分别导入HUVECs。免疫荧光法测定细胞内活性氧自由基(ROS)的生成。免疫印迹法检测细胞凋亡和抗凋亡因子。Anagliptin预孵育可保护HUVECs免受过氧化氢诱导的细胞凋亡。转染实验还表明，Anagliptin预孵育可抑制NADPH氧化酶4(NOX-4)过表达诱导的HUVEC的凋亡。免疫荧光显示，Anagliptin预孵育可抑制凋亡的HUVECs中ROS的形成。Anagliptin可抑制NOX-4介导的Bax、caspase-3、裂解caspase-3和Cyto C的过度表达，但上调Bcl2的蛋白水平。这些数据有助于我们更好地了解Anagliptin对细胞凋亡的影响，并将对寻找新的靶点来防止损伤后的内皮细胞凋亡具有重要意义。
Dipeptidyl peptidase IV (DPP-IV) inhibitors are novel oral anti-hyperglycemic agents. Here, the anti-apoptotic effect of Anagliptin in human umbilical vein endothelial cells (HUVECs) was evaluated. Cultured HUVECs were pre-incubated with Anagliptin, and then treated hydrongen peroxide (H2O2) to induce apoptosis. The apoptosis of HUVECs were detected by viability, LIVE/DEAD staining assay and flow cytometry assays. HUVECs were transfected with plasmid harboring human NADPH oxidases (NOX) 4 or an empty vector. The formation of reactive oxygen species (ROS) was measured by immunofluorescence. Apoptotic and anti-apoptotic factor were detected by Western Blot. Pre-incubation with Anagliptin protected HUVECs from H2O2induced apoptosis. The transfection assay also indicated that pre-incubation with Anagliptin inhibited the apoptosis of HUVECs induced by NADPH oxidase 4 (NOX-4) overexpression. Immunofluorescence demonstrated that pre-incubation with Anagliptin suppressed the formation of ROS in apoptotic HUVECs. Pre-incubation with Anagliptin inhibited NOX-4 mediated the Bax, caspase-3, cleave caspase-3 and Cyto C overexpression, but up-regulated the protein level of Bcl-2 in HUVECs. The data help us to better understand the effect of Anagliptin on apoptosis, and will be valuable in identifying new targets to prevent the endothelial cell apoptosis after injury.