RK002

Ginsenoside Rh 2 , a protopanaxadiol saponin from ginseng, has been reported to have strong anti-inflammatory activity. However, the concentration of ginsenoside Rh 2 is very low (>0.001%) in the total ginseng extracted, which is not enough for production despite its high pharmacological effects. Thus, in this study, we evaluated the anti-inflammatory effect of ginsenoside Rh 2 -mix (GRh 2 -mix) on lipopolysaccharide (LPS)-stimulated RAW 264.7 murine macrophage cells. From the high-performance liquid chromatography analysis, it was confirmed that the GRh 2 -mix was mainly composed of 20( S )-Rh 2 , 20( R )-Rh 2 , Rk 2 , and Rh 3 . The LPS-stimulated RAW 264.7 cells were treated with different concentrations of GRh 2 -mix (100, 200, 400, 500  μ g/mL). The cell counting kit-8 assay showed that the GRh 2 -mix treatment increased cell proliferation in LPS-stimulated RAW 264.7 murine macrophage cells. The GRh 2 -mix inhibited nitric oxide production in a dose-dependent manner, suggesting an anti-inflammatory effect. Furthermore, reverse transcription polymerase chain reaction and Western blot results also indicated that the GRh 2 -mix suppressed inflammatory genes such as iNOS , TNF - α , COX-2 , IL-1β , IL-6 , and NF - κB . In summary, these results suggest that the GRh 2 -mix exhibits anti-inflammatory activity via the downregulation of the NF- κ B pathway and has high efficiency with a simple production procedure.

Angiogenesis, a multi-step process which involves endothelial cell proliferation, adhesion, migration, and basement membrane (BM) degradation, is essential for tumor metastasis. Here we show that recombinant human prothrombin kringle-2 (rk-2) inhibited bovine capillary endothelial cell migration with an IC(50) (concentration for half maximal inhibition) of 38 nM and inhibited adhesion to extracellular matrix (ECM) proteins. Because tumor metastasis requires angiogenesis, we examined whether rk-2 could inhibit metastases induced by injection of B16F10 melanoma cells into mice. The results revealed that the metastatic tumors in mouse lung were markedly decreased in a dose-dependent manner and acute lung injury induced by B16F10 melanoma metastasis was diminished by systemic rk-2 treatment. In immunohistochemical analysis, rk-2 reduced expression of vascular endothelial growth factor, which is a potent angiogenic activator and neovascularization in the mouse lung. Also, rk-2 diminished the expression of matrix metalloproteinase-2 and -9 in the mouse lung which induces tumor metastasis and angiogenesis. These data suggest that inhibition of B16F10 melanoma metastasis by rk-2 was caused by inhibition of neovascularization and reduction of matrix metalloproteinase expression.