Screening anti-inflammatory components from Chinese traditional medicines using a peritoneal macrophage/cell membrane chromatography-offline-GC/MS method.
We report the development of an analytical method combining cell membrane chromatography (CMC) with gas chromatography/mass spectrometry (GC/MS). This was applied to the purification and identification of anti-inflammatory components from traditional Chinese medicines. The stationary phase of the CMC employed mouse peritoneal macrophage (PM) cell membranes. We investigated the performance of the PM/CMC-offline-GC/MS method using hydrocortisone (HC) and dexamethasone (DM) as standards. The method was then applied to the identification of anti-inflammatory components in extracts of Rhizoma Atractylodes macrocephala (RAM) and Rhizoma Atractylodes lancea Thunb DC (RALD). The major components from both species retained by CMC were identified as atractylenolide I (AO-I) by GC/MS. Competition experiments' results showed that AO-I and lipopolysaccharide (LPS) bound competitively to cell surface receptors while AO-I and HC had only partly overlapping binding sites on the PM membrane. In vitro experiments revealed that AO-I was able to inhibit LPS-induction of TNF-alpha, IL-1beta and NO production in a dose-dependent manner. IC50 values were 5.3microg/mL, 5.1microg/mL and 7.5microg/mL, respectively. The PM/CMC-offline-GC/MS method is an effective screening system for the rapid detection, enrichment, and identification of target components from complex samples.
Wang
Changhe
C
School of Medicine, Xi'an Jiaotong University, Xi'an, 710061, PR China.
He
Langchong
L
Wang
Nan
N
Liu
Fang
F
D000818:Animals; D031174:Atractylodes / Q000737:chemistry; D002462:Cell Membrane / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D002845:Chromatography / Q000379:methods*; D004365:Drugs, Chinese Herbal / Q000032:analysis*; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D006854:Hydrocortisone / Q000494:pharmacology; D053583:Interleukin-1beta / Q000378:metabolism; D007783:Lactones / Q000494:pharmacology; D017737:Macrophages, Peritoneal / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D009569:Nitric Oxide / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000378:metabolism; D015203:Reproducibility of Results; D027343:Rhizome / Q000737:chemistry; D012717:Sesquiterpenes / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D004365:Drugs, Chinese Herbal; D053583:Interleukin-1beta; D007783:Lactones; D011956:Receptors, Cell Surface; D012717:Sesquiterpenes; D014409:Tumor Necrosis Factor-alpha; C424804:atractylenolide I; D009569:Nitric Oxide; D006854:Hydrocortisone