Inhibition of Lipopolysaccharide-Induced Apoptosis by Cilostazol in Human Umbilical Vein Endothelial Cells

Abstract

This work describes the pharmacological inhibition by cilostazol and its metabolites, OPC-13015 and OPC-13213, of the apoptosis in the human umbilical vein endothelial cells (HUVECs) damaged by lipopolysaccharide (LPS) in comparison with its analog, cilostamide. Cilostazol and OPC-31213 caused a significant suppression of cell death induced by LPS (1 g/ml) in a concentration-dependent manner but a modest suppression by cilostamide and OPC-13015. These compounds potently inhibited the 5,5-dimethyl-1-pyrroline-1-oxide (DMPO)/ ⅐ OH adduct formation and significantly reduced the increased intracellular reactive oxygen species (ROS) and tumor necrosis factor-␣ (TNF-␣) production induced by LPS (1 g/ml). An apoptotic death of HUVECs by 1 g/ml LPS (DNA ladders on electrophoresis) was strongly suppressed by all these compounds. Incubation with LPS caused a marked decrease in Bcl-2 protein, which was significantly reversed by cilostazol and its analogs. The greatly increased Bax protein expression and cytochrome c release by LPS were, in contrast, suppressed by cilostazol and, to a lesser degree, by others. In conclusion, cilostazol and its analogs exert a strong protection against apoptotic cell death by scavenging hydroxyl radicals and intracellular ROS with reduction in TNF-␣ formation and by increasing Bcl-2 protein expression and decreasing Bax protein and cytochrome c release.

Cilostazol protects rat chondrocytes against nitric oxide–induced apoptosis in vitro and prevents cartilage destruction in a rat model of osteoarthritis

“…Previous studies (20,21) have shown that cilostazol reverses decreases in Bcl-2 protein and increases in Bax protein production and cytochrome c release. The maxi-K channel opening-coupled upregulation and down-regulation of PTEN phosphorylation with resultant increases in Akt and CREB phosphorylation and increased Bcl-2 protein levels have also been demonstrated (12).…”

“…The recent observation that cilostazol inhibits apoptosis under several circumstances (12,(20)(21)(22) raises the interesting possibility that it can be considered as a therapy for diseases associated with excess apoptosis. Although apoptotic chondrocyte death is not always a widespread phenomenon in cartilage aging or OA cartilage degeneration (38), obviously any kind of cell death is detrimental to the tissue.…”

“…In addition, it seems to reduce inflammation and inhibit MMP expression by increasing AMP and inhibiting the p38/ JNK pathway (19). Previous studies have demonstrated that cilostazol inhibits apoptosis under several circumstances (12,(20)(21)(22). In those studies, cilostazol was shown to prevent lipoprotein particle-induced apoptosis in human umbilical vein endothelial cells (HUVECs) (22), lipopolysaccharide-induced apoptosis in HUVECs (20), and ischemic injury-induced apoptosis in neural cells (12,21).…”

“…Previous studies have demonstrated that cilostazol inhibits apoptosis under several circumstances (12,(20)(21)(22). In those studies, cilostazol was shown to prevent lipoprotein particle-induced apoptosis in human umbilical vein endothelial cells (HUVECs) (22), lipopolysaccharide-induced apoptosis in HUVECs (20), and ischemic injury-induced apoptosis in neural cells (12,21). Notably, protein kinase CK2 (PKCK2) was reported to be involved in the prevention of apoptosis by cilostazol (12,23).…”

Cilostazol, um inibidor da fosfodiesterase III: perspectivas futuras na aterosclerose

“…Cilostazol increased the NO expression in cell cultures [67][68][69][70] and rats 71 , positively altered oxidative stress 53,55 , inhibited apoptosis induced by lipopolysaccharides; it diminished BAX gene levels and increased Bcl-2 gene levels, in cultures of endothelial cells taken from a human umbilical cord 72 . It also diminished cerebral stroke in association with apoptosis inhibition and oxidative cellular death in rats submitted to focal cerebral ischemia 73 .…”