Human umbilical vein endothelial cell proliferation and migration of Kot Chulalumpa in Phikud Navakot and preliminary chemical profile

Abstract

Phikud Navakot has been recognized as a part of Yahom Navakot for the treatment of circulatory disorder. The present study aimed to evaluate the effects of the ethanol and water extracts of crude drugs in Phikud Navakot on proliferation and migration of human umbilical vein endothelial ECV304 cells. Quality control parameters of nine crude drugs were examined according to Thai Herbal Pharmacopoeia before extraction with either 50% ethanol or water. Treatment of the cells with the ethanol extract of Kot Chulalumpa (Artemisia pallens, APES) for 24 h significantly increased cell proliferation using MTT reduction assay. Among nine herbal extracts, APES exhibited the highest activity. An in vitro scratch assay showed that treatment with APES for 6 h under serum-free condition significantly promoted endothelial wound closure in a concentration-dependent manner via enhancing cell migration. Furthermore, DCFH-DA assay revealed that APES significantly decreased intracellular reactive oxygen species level via activating the activity of superoxide dismutase (SOD) but not glutathione peroxidase and catalase. APES significantly stimulated both cyclooxygenase (COX) -1 and COX-2 activities without affecting intracellular nitric oxide level using 2,3-diaminonaphthalene assay. High performance liquid chromatography profile revealed the presence of flavonoids such as apigenin, eriodictyol, luteolin, quercetin and rutin, which might be used as chemical markers for identification of APES. Thus, the ethanol extract of A. pallens enhanced endothelial cell proliferation and migration via activating the activities of SOD and COX in a nitric oxide-independent pathway, leading to its potential role in angiogenesis for the treatment of circulatory disorder of Phikud Navakot.