Preparative isolation and validation of compounds from Rhus verniciflua by high-speed counter-current chromatography, and their relevance for treatment of eye-related diseases
- Preparative isolation and validation of compounds from Rhus verniciflua by high-speed counter-current chromatography, and their relevance for treatment of eye-related diseases
- 최순정; 이민영; 조형; 임순성; 정상훈
- Rhus verniciflua; Neuroprotection; High-speed counter-current chromatography (HSCCC); Retinal ganglion cell-5 (RGC-5)
- Issue Date
- 2012 한국응용생명화학회 천연물분과 심포지엄
- In the present study, we isolated four flavonoids from Rhus verniciflua by using high-speed counter-current chromatography (HSCCC). Their bioactivities were assessed using cell viability assays, reactive oxygen species assays, western blot analysis and lipid peroxidation assays.
Four flavonoids from R. verniciflua were successfully isolated using HSCCC with a two-phase solvent system composed of n-hexane/ethyl acetate/methanol/water (3.5:5:3.5:5, v/v). From 1.5 g of R. verniciflua extract, we successfully used this method to separate 252.1 mg of fustin at a purity of > 93.09%, 51.2 mg of fisetin at a purity of > 95.45%, 39.7 mg of sulfuretin at a purity of > 95.17%, and 10.7 mg of butein at a purity of > 95.01%. Their structures were identified by ESI-MS, 1H-NMR, 13C-NMR, and 2D NMR, and were confirmed by direct comparison of spectral data from literature.
The extract and compounds from R. verniciflua were found to significantly attenuate the toxic effects of hydrogen peroxide (H2O2) on a transformed retinal ganglion cell line (RGC-5 cells), and to both reduce the levels of reactive oxygen species present and lessen the up-regulation of apoptotic proteins such as cleaved PAPR, cleaved caspase-3, and cleaved caspase-9.
These results demonstrate that R. verniciflua has protective effects on oxidative stress-induced retinal degeneration.
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