title: Combined prescription (OAH19T) of Aralia cordata Thunb and Cimicifuga heracleifolia Komar and its major compounds inhibit matrix proteinases and vascular endothelial growth factor through the regulation of mitogen-activated protein kinase pathway.
abstract: BACKGROUND
  OAH19T, a new herbal extract from a mixture of Aralia cordata Thunb and Cimicifuga heracleifolia, is traditionally used for the treatment of arthritis in far East Asia. To investigate the chondroprotective effects of OAH19T on osteoarthritis was examined and compared with its major compounds pimaradienoic acid (PA) and ferulic acid (FA) of human osteoarthritis (OA) chondrocytes.
  
  
  METHODS
  Chondrocytes, alone or in the presence of IL-1β, were cultured with or without OAH19T, PA or FA (10, 20, 40 μg/ml). The release of sulfated glycosaminoglycan (GAG) was measured by colorimetric assay using 1,9-dimethylmethylene blue (DMB) reagent from the cultured media. The level of aggrecanases and VEGF was measured by reverse transcription polymerase chain reaction (RT-PCR). The expression of MMP-1 and MMP-3 analyzed by real time RT-PCR and enzyme-linked immunosorbent assay (ELISA). The phosphorylation of mitogen-activated protein kinases was performed by immunoblotting in OA chondrocytes. The proliferation was examined by the BrdU assay.
  
  
  RESULTS
  OAH19T markedly inhibited the release of proteoglycan and the degradation of aggrecan, in a dose-dependent manner in OA chondrocytes. OAH19T also inhibited the level of aggrecanase-1, aggrecanase-2, MMP-1, MMP-3, and VEGF in OA chondrocytes. PA and FA also inhibited the level of aggrecanase-2, MMP-3 and VEGF, while did not significantly affect the levels of aggrecanase-1, MMP-3 in OA chondrocytes. OAH19T exhibited the down-regulation of p38 MAP kinase unlike PA and FA in OA chondrocytes without cytotoxicity. In addition, p38 inhibitor SB203580 abolished the antiproliferative activity and proteoglycan degradation by OAH19T, while had no effect by PA or FA.
  
  
  CONCLUSIONS
  OAH19T have shown the chondroprotective effect by inhibiting cell proliferation, expression of cartilage-specific matrix proteinases and release of VEGF, but bigger than PA or FA, through down-regulation of p38 MAP kinase in human OA chondocyte. These results provide pharmacological basis for use in treatment of OA.
authors.affiliation: Oriental Medicine Research Center for Bone & Joint Disease, Kyung Hee University, 149, Sangil-dong, Gangdong-gu, Seoul 134-727, Republic of Korea.
authors.forename: Jeong-Eun
authors.initials: JE
authors.lastname: Huh
authors.forename: Yae-Ji
authors.initials: YJ
authors.lastname: Shin
authors.forename: Yong-Hyeon
authors.initials: YH
authors.lastname: Baek
authors.forename: Jae-Dong
authors.initials: JD
authors.lastname: Lee
authors.forename: Do-Young
authors.initials: DY
authors.lastname: Choi
authors.forename: Dong-Suk
authors.initials: DS
authors.lastname: Park
mesh_terms: D027983:Aralia; D001483:Base Sequence; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000187:drug effects; D031885:Cimicifuga; D017931:DNA Primers; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D061965:Matrix Metalloproteinase Inhibitors; D011480:Protease Inhibitors / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D042461:Vascular Endothelial Growth Factor A / Q000037:antagonists & inhibitors*
chemical_list: D017931:DNA Primers; D061965:Matrix Metalloproteinase Inhibitors; D011480:Protease Inhibitors; D042461:Vascular Endothelial Growth Factor A