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Celastrol,is an active compound extracted from the root bark of the traditional Chinese medicine Tripterygium wilfordii Hook F.In this study,we investigated the effect of celastrol on LPS-activated LP-1 human multiple myeloma cell-induced angiogenesis,and identified its molecular mechanism of action.Human umbilical vein endothelial (HUVEC) migration was tested using a wound-healing assay.HUVEC invasion was assayed using a Transwell chamber.Cell surface TLR4 expression was analyzed by flow cytometry.Angiogenic factor VEGF level was quantified by LUMINEX and protein expression was analyzed by Western blot.NF-κ B nuclear translocation was observed by fluorescence microscopy.Celastrol inhibited LPS-stimulated LP-1 human multiple myeloma-induced HUVECs cell migration and invasion in a concentration-dependent manner.Wound diameters increased by 72.9%,165.4% and 246.2% at 0.025,0.05 and 0.1 μ M,respectively,compared with LPS alone.A 45 to 74% inhibition of LPS-dependent cell invasion was achieved in the presence of 0.025 to O.1 μ M celastrol.Celastrol significantly downregulated LPS induced TLR4 expression and inhibited LPS-induced VEGF secretion in LP-1 cells.VEGF levels decreased by 64.8%,84.4% and 92.9% after co-exposure to celastrol at 0.025,0.05 and 0.1 μ M,compared with the LPS alone.Celastrol also inhibited the IKK/NF-kB pathway induced by LPS.Protein levels of NF-κ B p65,IKKa and I κ B-α decreased in a dose-dependent manner after co-exposure to celastrol.Celastrol also blocked nuclear translocation of the p65 subunit.These results suggest that celastrol inhibits lipopolysaccharide-induced angiogenesis by suppressing TLR4-triggered NF-kappa B activation.