Genipin inhibits endothelial exocytosis via nitric oxide in cultured human umbilical vein endothelial cells

Abstract

Aim: Exocytosis of endothelial Weibel-Palade bodies, which contain von Willebrand factor (VWF), P-selectin and other modulators, plays an important role in both inflammation and thrombosis. The present study investigates whether genipin, an aglycon of geniposide, inhibits endothelial exocytosis. Methods: Human umbilical vein endothelial cells (HUVECs) were isolated from umbilical cords and cultured. The concentration of VWF in cell supernatants was measured using an ELISA Kit. P-selectin translocation on the cell surface was analyzed by cell surface ELISA. Cell viability was measured using a Cell Counting Kit-8. Mouse bleeding times were measured by amputating the tail tip. Western blot analysis was used to determine the amount of endothelial nitric oxide synthase (eNOS) and phospho-eNOS present. Nitric oxide (NO) was measured in the cell supernatants as nitrite using an NO Colorimetric Assay. Results: Genipin inhibited thrombin-induced VWF release and P-selectin translocation in HUVECs in a dose-and timedependent manner. The drug had no cytotoxic effect on the cells at the same doses that were able to inhibit exocytosis. The functional study that demonstrated that genipin inhibited exocytosis in vivo also showed that genipin prolonged the mouse bleeding time. Furthermore, genipin activated eNOS phosphorylation, promoted enzyme activation and increased NO production. L-NAME, an inhibitor of NOS, reversed the inhibitory effects of genipin on endothelial exocytosis. Conclusion: Genipin inhibits endothelial exocytosis in HUVECs. The mechanism by which this compound inhibits exocytosis may be related to its ability to stimulate eNOS activation and NO production. Our findings suggest a novel antiinflammatory mechanism for genipin. This compound may represent a new treatment for inflammation and/or thrombosis in which excess endothelial exocytosis plays a pathophysiological role.

Geniposide inhibits high glucose-induced cell adhesion through the NF-κB signaling pathway in human umbilical vein endothelial cells

“…We also found that genipin, the aglycone of geniposide, inhibits endothelial exocytosis in HUVECs by stimulating nitric oxide production [15] . However, whether geniposide prevents high glucose-induced vascular injury was not investigated.…”

Genipin, a constituent of Gardenia jasminoides Ellis, induces apoptosis and inhibits invasion in MDA-MB-231 breast cancer cells

“…Genipin exerts anti-inflammatory, anti-angiogenic, antiproliferative and hepatoprotective activities in several cell lines including murine macrophage cells, human umbilical vein endothelial cells, human prostate cancer cells, and human hepatocarcinoma cells (2)(3)(4)(5). Genipin has been shown to inhibit growth of human leukemia and prostate cancer cells (5,6).…”

Apoptosis induced by genipin in human leukemia K562 cells: involvement of c-Jun N-terminal kinase in G2/M arrest

“…www.nature.com/aps Feng Q et al Acta Pharmacologica Sinica npg cells (HUVECs), suppresses vascular and endothelial cell infl ammatory activation, and may target acute infl ammatory events [12] . Notably, genipin has been shown to induce dosedependent apoptosis in FaO rat hepatoma cells, human hepatocarcinoma Hep3B cells [13] and PC3 human prostate cancer cells [14] .…”