Prevention of Lipopolysaccharide-Induced Apoptosis by (2 S,3 S,4 R)-N″-Cyano-N-(6-amino-3,4-dihydro-3-hydroxy-2-methyl-2-dimethoxymethyl-2 H-benzopyran-4-yl)-N′-benzylguanidine, a Benzopyran Analog, in Endothelial Cells

• Published in: The Journal of pharmacology and experimental therapeutics 2002-02, Vol.300 (2), p.535-542
• Main Authors: Kim, Ki Young, Kim, Byeong Gee, Kim, Sun-Ok, Yoo, Sung-Eun, Kwak, Yong-Geun, Chae, Soo-Wan, Hong, Ki Whan
• Format: Article
• Language: English
• Subjects: alpha-Tocopherol - pharmacology; Apoptosis - drug effects; bcl-2-Associated X Protein; Blotting, Western; Cell Survival - drug effects; Cells, Cultured; Cytochrome c Group - biosynthesis; Cytochrome c Group - genetics; DNA Fragmentation - drug effects; Endothelium, Vascular - cytology; Endothelium, Vascular - drug effects; Free Radical Scavengers - pharmacology; Gene Expression Regulation - drug effects; Genes, bcl-2 - drug effects; Humans; Hydrogen Peroxide - metabolism; Lipopolysaccharides - antagonists & inhibitors; Lipopolysaccharides - toxicity; Mitochondria - drug effects; Mitochondria - enzymology; Proto-Oncogene Proteins - biosynthesis; Proto-Oncogene Proteins - genetics; Proto-Oncogene Proteins c-bcl-2; Reactive Oxygen Species - metabolism; Tumor Necrosis Factor-alpha - metabolism
• ISSN: 0022-3565
• DOI: 10.1124/jpet.300.2.535

This study describes the antiapoptotic action of (2 S,3 S,4 R)-N"-cyano-N-(6-amino-3,4-dihydro-3-hydroxy-2-methyl-2-dimethoxymethyl-2 H-benzopyran-4-yl)-N′-benzylguanidine (KR-31378), a novel benzopyran analog, in human umbilical vein endothelial cells (HUVECs) in comparison with its acetylated metabolite, (2 S,3 S,4 R)-N"-cyano-N-(6-acetylamino-3,4-dihydro-3-hydroxy-2-methyl-2-dimethoxymethyl-2 H-benzopyran-4-yl)-N′-benzylguanidine (KR-31612), and with α-tocopherol. Exposure of HUVECs to lipopolysaccharide (LPS) (1 μg/ml) induced time- and concentration-dependent cytotoxicity and oligonucleosomal DNA fragmentation. KR-31378, KR-31612, and α-tocopherol potently suppressed LPS-induced cell death in association with significant reduction in the intracellular reactive oxygen species (ROS) and tumor necrosis factor-α (TNF-α) that are stimulated by LPS. KR-31378 more effectively protected HUVECs from LPS-induced DNA fragmentation and was more effective in peroxyl radical scavenging than α-tocopherol. Incubation with LPS markedly decreased the Bcl-2 level, which was totally reversed by KR-31378 and to a lesser degree by KR-31612 and by α-tocopherol. In contrast, the greatly increased Bax protein and cytochrome c release stimulated by LPS were markedly suppressed by KR-31378 and by KR-31612, and to a lesser degree by α-tocopherol. Taken together, KR-31378 strongly inhibited cell death in HUVECs in association with antiapoptotic effects, which were accompanied by up-regulation of Bcl-2 protein expression and down-regulation of Bax protein and suppression of cytochrome c release. KR-31378 also showed the properties to scavenge the intracellular ROS and peroxyl radicals, and to reduce the TNF-α production induced by LPS.