Preparative isolation and purification of neuroprotective compounds from Rhus verniciflua by high speed counter-current chromatography

Title
Preparative isolation and purification of neuroprotective compounds from Rhus verniciflua by high speed counter-current chromatography
Authors
최순정이민영조형임순성정상훈
Keywords
Rhus verniciflua; neuroprotection; HSCCC; RGC-5; high-speed counter-current chromatography; retinal ganglion cell-5
Issue Date
2012-04
Publisher
Biological & pharmaceutical bulletin
Citation
VOL 35, NO 4, 559-567
Abstract
In the present study, extracts from Rhus verniciflua were demonstrated to significantly attenuate the negative effects of hydrogen peroxide (H2O2) on transformed retinal ganglion cell line (RGC-5 cells), indicating that they may be protective against oxidative stress-induced retinal degeneration. The inclusion of R. verniciflua in the culture was found to both reduce the levels of reactive oxygen species (ROS) present and lessen the up-regulation of apoptotic proteins such as cleaved poly(ADP-ribose) polymerase, cleaved caspase-3, and cleaved caspase-9. Active compounds were also successfully isolated from R. verniciflua using high-speed counter-current chromatography (HSCCC) with a two-phase solvent system composed of n-hexane–ethyl acetate– methanol–water (3.5 : 5 : 3.5 : 5, v/v). Using this method, we successfully separated 252.1 mg of fustin at a purity of over 93.09%, 51.2 mg of fisetin at a purity of over 95.45%, 39.7 mg of sulfuretin at a purity of over 95.17%, and 10.7 mg of butein at a purity of over 95.01% from 1.5 g of R. verniciflua extract. The chemical structures of these compounds were elucidated by chemical and spectral analyses. There isolated compounds also significantly attenuated the negative effects of H2O2 on RGC-5 cells. Results therefore suggest that, due to its anti-oxidative and anti-apoptotic effects, R. verniciflua could be used as a lead substance for the treatment of retinal diseases such as glaucoma.
URI
http://pubs.kist.re.kr/handle/201004/43730
ISSN
09186158
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