7-Difluoromethyl-5, 4′-Dimethoxygenistein Reverses LPC-Induced Apoptosis of HUVE-12 Cells Through Regulating Mitochondrial Apoptosis Pathway
7-Difluoromethyl-5, 4′-Dimethoxygenistein Reverses LPC-Induced Apoptosis of HUVE-12 Cells Through Regulating Mitochondrial Apoptosis Pathway
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DOI：
10.2174/157436240901140924105409
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发表时间：
2014-01-01
影响因子：
--
通讯作者：
Zhang, Yong
中科院分区：
文献类型：
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作者：
Liu, Shiwei;Li, Lesai;Zhang, Yong
7-二氟甲基-5，4 '-二甲氧基异黄酮（DFMG）是由染料木黄酮经二氟甲基化和烷基化反应制得的一种新的活性化合物。本研究探讨了DFMG对溶血磷脂酰胆碱（LPC）诱导的人脐静脉内皮细胞-12（HUVEC-12）线粒体凋亡途径的抑制作用。用溶血磷脂酰胆碱（LPC）建立HUVEC-12细胞凋亡模型。ANNEXIN V/PI双染流式细胞仪检测各组细胞凋亡率。用流式细胞仪测定线粒体膜电位（MMP）的平均荧光强度（MFI）。Western blotting检测细胞色素C（CytC）、p-JNK和PARP的表达水平。caspase-3和caspase-9活性采用caspase比色法测定。与对照组相比，DFMG可显著降低LPC诱导的细胞凋亡率、MMP的MFI和活性氧（ROS）的产生。此外，DFMG对LPC处理的HUVEC-12细胞中CytC、P-JNK、PARP的表达及caspase-3、caspase-9的活性均有抑制作用。DFMG可拮抗LPC诱导的HUVE-12细胞凋亡。其机制可能与抑制线粒体凋亡途径有关。
7-Difluoromethyl- 5, 4'-dimethoxygenistein (DFMG), prepared by the difluoromethylation and alkylation of Genistein, is a new active chemical entity. In this study, we investigated the inhibitory effect of DFMG on Lysophosphatidyl choline (LPC)-induced apoptosis of human umbilical vein endothelial cells-12 (HUVEC-12) through mitochondrial apoptosis pathway. Lysophosphatidyl choline (LPC) was used to build apoptosis model in HUVEC-12 cells. The apoptosis rate in each group was determined by using flow cytometry of ANNEXIN V/PI staining. The mean ffluorescence intensity (MFI) of mitochondria mmembrane ppotential (MMP) was determined by using flow cytometry. The expression levels of cytochrome C (CytC), p-JNK and PARP were detected by performing western blotting assay. The activities of caspase-3 and caspase-9 were detected by a colorimetric method with caspase colorimetric assay kits. As compared with the control group, DFMG notably decreased the apoptosis rate, the MFI of the MMP, and the generation of Reactive Oxygen Species (ROS) induced by LPC. In addition, DFMG had suppressive effects on the expressions of CytC, P-JNK, and PARP, as well as on the activities of caspase-3 and caspase-9 in LPC-treated HUVEC-12 cells. DFMG can antagonize LPC-induced apoptosis of HUVE-12 cells. The underlying mechanism may be involved in the inhibition of mitochondrial apoptosis pathway.