Screening of immunomodulatory components in Yu-ping-feng-san using splenocyte binding and HPLC.
Yu-ping-feng-san (YPFS) is a widely used immunomodulatory herbal medication used in traditional Chinese medicine, but the active molecules remain obscure. To screen for bioactive components we combined splenocyte binding with high performance liquid chromatography (SB-HPLC). After enrichment by splenocyte binding, two YPFS components (C1 and C2) were analyzed by HPLC. Compound C2 was identified as linoleic acid (LA) based on UV absorption and mass spectrometry. Silica gel chromatography was used to purify compound C1 from Radix Saposhnikoviae, a major constituent of YPFS. This allowed identification of the molecule as panaxynol (PAN) based on EI-MS and NMR spectrometry. Bioassay in vitro demonstrated that PAN significantly inhibited splenocyte proliferation induced by concanavalin A (ConA) in a concentration-dependent manner, whereas LA had no significant effect on splenocyte proliferation. In vivo, PAN was found to attenuate allergic contact dermatitis in a mouse model of delayed-type hypersensitivity (DTH), a pharmacological activity not previously reported for this molecule. It is suggested that PAN contributes to the anti-DTH effects of YPFS. SB-HPLC provides a rapid and efficient method for the identification of potential immunomodulatory components in traditional Chinese medicines.
Hong
Min
M
Jiangsu Key Laboratory for Traditional Chinese Medical Formulae Research, Nanjing University of Chinese Medicine, Nanjing 210046, China.
Wang
Xin-zhi
XZ
Wang
Liang
L
Hua
Yong-qing
YQ
Wen
Hong-mei
HM
Duan
Jin-ao
JA
D000818:Animals; D049109:Cell Proliferation; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003208:Concanavalin A / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004365:Drugs, Chinese Herbal / Q000737:chemistry* / Q000494:pharmacology; D007155:Immunologic Factors / Q000627:therapeutic use; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008297:Male; D013058:Mass Spectrometry / Q000379:methods; D008516:Medicine, Chinese Traditional / Q000379:methods; D051379:Mice; D011485:Protein Binding; D013154:Spleen / Q000166:cytology* / Q000187:drug effects; D014466:Ultraviolet Rays
D004365:Drugs, Chinese Herbal; D007155:Immunologic Factors; C506362:yu ping feng san; D003208:Concanavalin A