title: Characterization and immunostimulating effects on murine peritoneal macrophages of a novel protein isolated from Panax quinquefolius L.
abstract: BACKGROUND
  Panax quinquefolius L. has been used as a proverbial tonic in oriental countries for hundreds of years. It is used as a traditional medicinal herb to nourish vitality.
  
  
  OBJECTIVE
  The purpose of our study was to inquiry the activation effects on murine peritoneal macrophages of a novel protein separated from the roots of Panax quinquefolius L.
  
  
  METHODS
  In our work, a novel protein of the roots of American ginseng (AGNP) was separated and purified from the roots of Panax quinquefolius L. The characteristic was investigated with SDS-PAGE, high pressure gel filtration chromatography (HPGFC) and matrix-assisted laser desorption ionization/time-of-flight mass (MALDI-TOF-MS) spectrometry method. The method of neutral red was carried out to investigate the phagocytosis of peritoneal macrophages. And Griess method and colorimetry were executed to detect the level of nitric oxide and iNOS activity respectively. Tumor necrosis factor-α and interleukin-6 were analyzed by enzyme linked immunosorbent assay (ELISA).
  
  
  RESULTS
  Our results demonstrated that the subunit molecular weight of AGNP determined by SDS-PAGE was 15kD and the content of proteins determined by Bradford assay was 2.31mg/mL. The molecular weight of the AGNP was15, 114Da both of electrophoresis and MS purity. And the result of HPGFC showed that the molecular weight of AGNP was 31,086Da, Immunological studied indicated that AGNP could conspicuously increase phagocytosis of macrophages, facilitate the nitric oxide production, Tumor necrosis factor-α and interleukin-6 production. What is more, AGNP dose-dependently stimulated NO formation through the up-regulation of iNOS activity.
  
  
  CONCLUSIONS
  In conclusion, AGNP had good immunoregulatory effects supporting the traditional claims and may provide a valuable therapeutic strategy to promoting immune function and metabolism.
authors.affiliation: College of Pharmacy，Changchun University of Chinese Medicine, Changchun, Jilin, 130117, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Qi
authors.affiliation: College of Pharmacy，Changchun University of Chinese Medicine, Changchun, Jilin, 130117, China.
authors.forename: Shan
authors.initials: S
authors.lastname: Wang
authors.affiliation: College of Pharmacy，Changchun University of Chinese Medicine, Changchun, Jilin, 130117, China.
authors.forename: Qiu
authors.initials: Q
authors.lastname: Wang
authors.affiliation: The Affiliated Hospital to Changchun University of Chinese Medicine, Changchun, Jilin 130021, China.
authors.forename: He
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Center of Chinese Medicine and Bio-Engineering Research, Changchun University of Chinese Medicine, Changchun, Jilin, 130117, China.
authors.forename: Xue-Yuan
authors.initials: XY
authors.lastname: Bai
authors.affiliation: College of Pharmacy，Changchun University of Chinese Medicine, Changchun, Jilin, 130117, China.
authors.forename: Hui-Nan
authors.initials: HN
authors.lastname: He
authors.affiliation: College of Pharmacy，Changchun University of Chinese Medicine, Changchun, Jilin, 130117, China.
authors.forename: Wei-Jie
authors.initials: WJ
authors.lastname: Sun
authors.affiliation: College of Pharmacy，Changchun University of Chinese Medicine, Changchun, Jilin, 130117, China.
authors.forename: Li
authors.initials: L
authors.lastname: Liu
authors.affiliation: Center of Chinese Medicine and Bio-Engineering Research, Changchun University of Chinese Medicine, Changchun, Jilin, 130117, China. Electronic address: cnzhaodaqing@126.com.
authors.forename: Da-Qing
authors.initials: DQ
authors.lastname: Zhao
mesh_terms: D000276:Adjuvants, Immunologic / Q000302:isolation & purification / Q000494:pharmacology*; D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D016207:Cytokines / Q000096:biosynthesis; D017737:Macrophages, Peritoneal / Q000187:drug effects* / Q000276:immunology; D008297:Male; D008813:Mice, Inbred ICR; D008970:Molecular Weight; D009569:Nitric Oxide / Q000096:biosynthesis; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism; D005894:Panax / Q000737:chemistry*; D010587:Phagocytosis / Q000187:drug effects* / Q000276:immunology; D010940:Plant Proteins / Q000302:isolation & purification / Q000494:pharmacology*; D018517:Plant Roots / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D000276:Adjuvants, Immunologic; D016207:Cytokines; D010940:Plant Proteins; D009569:Nitric Oxide; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse
keywords: Immunostimulation; Macrophages; Novel protein; Panax quinquefolium