title: Preparative isolation and purification of neuroprotective compounds from Rhus verniciflua by high speed counter-current chromatography.
abstract: In the present study, extracts from Rhus verniciflua were demonstrated to significantly attenuate the negative effects of hydrogen peroxide (H(2)O(2)) 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 H(2)O(2) 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.
authors.affiliation: Functional Food Center, Korea Institute of Science and Technology, Gangneung Institute, Republic of Korea.
authors.forename: Soon Jung
authors.initials: SJ
authors.lastname: Choi
authors.forename: Min Young
authors.initials: MY
authors.lastname: Lee
authors.forename: Hyoung
authors.initials: H
authors.lastname: Jo
authors.forename: Soon Sung
authors.initials: SS
authors.lastname: Lim
authors.forename: Sang Hoon
authors.initials: SH
authors.lastname: Jung
mesh_terms: D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D003377:Countercurrent Distribution; D006861:Hydrogen Peroxide / Q000009:adverse effects; D015227:Lipid Peroxidation / Q000187:drug effects; D008297:Male; D018696:Neuroprotective Agents / Q000302:isolation & purification / Q000494:pharmacology*; D016877:Oxidants / Q000009:adverse effects; D024301:Plant Bark / Q000737:chemistry; D010936:Plant Extracts / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000378:metabolism; D012165:Retinal Ganglion Cells / Q000187:drug effects* / Q000378:metabolism; D029045:Rhus / Q000737:chemistry*
chemical_list: D018696:Neuroprotective Agents; D016877:Oxidants; D010936:Plant Extracts; D017382:Reactive Oxygen Species; D006861:Hydrogen Peroxide