title: Anti-inflammatory effects of the extract of Gnaphalium affine D. Don in vivo and in vitro.
abstract: BACKGROUND
  Gnaphalium affine D. Don (GA) has been traditionally used as a medicinal herb in China for the treatment of many ailments including rheumatoid arthritis. However, the anti-arthritic mechanism of GA has still not been demonstrated. This study aims to reveal the anti-inflammatory activity and anti-arthritic mechanism of ethanol extract of G. affine D. Don.
  
  
  METHODS
  Anti-inflammatory potential of GA was analyzed in vivo in carrageenan induced mice paw edema (acute study). Also, in vivo study was applied in collagen-induced arthritis (CIA) rats. In vitro experiments for analyzing the anti-inflammatory potential of GA were performed on rat alveolar macrophages cell line (NR8383). Analysis of nitric oxide release in NR8383 cells was done by Griess reaction. RT-PCR and western blotting experiment was performed to analyze the expression of phosphorylated p65 and IκBα/β-actin in NF-κB pathway. The production of TNF-α, IL-1β, and COX-2 in NR8383 cells were measured by enzyme-linked immunosorbent assay. The chemical profile of GA was analyzed by HPLC-VWD.
  
  
  RESULTS
  GA significantly reduced the paw volume in carrageenan induced rat paw edema rat at different doses (300 and 600 mg/kg), compared with the standard indomethacin treatment. In CIA, GA can obviously ameliorate the inflammatory symptom, including cytokine, histological symptom and paw swelling. In the vitro study, GA was able to reduce the nitric oxide (NO) levels in NR8383 cells that had been stimulated with lipopolysaccharide (LPS). The level of TNF-α, IL-1β, and COX-2 was also decreased with GA treatment in NR8383 cells that had been stimulated with lipopolysaccharide (LPS). Interestingly, GA was found to decrease the level of phosphorylated p65 and IκBα in NR8383 cells. Fifteen compounds were identified by HPLC-VWD with the reference substances and verified by LC-MS.
  
  
  CONCLUSIONS
  The results of the experiment scientifically validated its traditional use in inflammatory conditions.
authors.affiliation: Department of Identification of Traditional Chinese Medicine, School of Pharmacy, Second Military Medical University, Shanghai 200433, PR China.
authors.forename: Doudou
authors.initials: D
authors.lastname: Huang
authors.affiliation: Department of Identification of Traditional Chinese Medicine, School of Pharmacy, Second Military Medical University, Shanghai 200433, PR China.
authors.forename: Yanhong
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of Pharmacy, Changzheng Hospital, Shanghai 200003, PR China.
authors.forename: Wansheng
authors.initials: W
authors.lastname: Chen
authors.affiliation: Department of Pharmacology, China State Institute of Pharmaceutical Industry, Shanghai 200040, PR China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Identification of Traditional Chinese Medicine, School of Pharmacy, Second Military Medical University, Shanghai 200433, PR China.
authors.forename: Fengyan
authors.initials: F
authors.lastname: Yao
authors.affiliation: Department of Identification of Traditional Chinese Medicine, School of Pharmacy, Second Military Medical University, Shanghai 200433, PR China.
authors.forename: Dan
authors.initials: D
authors.lastname: Xue
authors.affiliation: Department of Identification of Traditional Chinese Medicine, School of Pharmacy, Second Military Medical University, Shanghai 200433, PR China. Electronic address: sssnmr@163.com.
authors.forename: Lianna
authors.initials: L
authors.lastname: Sun
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology* / Q000627:therapeutic use*; D001169:Arthritis, Experimental / Q000097:blood / Q000188:drug therapy / Q000473:pathology; D002351:Carrageenan; D002460:Cell Line; D004487:Edema / Q000188:drug therapy / Q000473:pathology; D005260:Female; D005528:Foot / Q000473:pathology; D036622:Gnaphalium; D053583:Interleukin-1beta / Q000097:blood / Q000378:metabolism; D007719:Knee Joint / Q000187:drug effects / Q000473:pathology; D008297:Male; D008813:Mice, Inbred ICR; D016328:NF-kappa B / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D008517:Phytotherapy; D010936:Plant Extracts / Q000494:pharmacology* / Q000627:therapeutic use*; D017208:Rats, Wistar; D014409:Tumor Necrosis Factor-alpha / Q000097:blood / Q000378:metabolism
chemical_list: D000893:Anti-Inflammatory Agents; D053583:Interleukin-1beta; D016328:NF-kappa B; D010936:Plant Extracts; D014409:Tumor Necrosis Factor-alpha; D009569:Nitric Oxide; D002351:Carrageenan
keywords: Carrageenan induced mice paw edema; Gnaphalium affine D. Don; Inflammation; Lipopolysaccharide