title: Effect of a Herbal-Leucine mix on the IL-1β-induced cartilage degradation and inflammatory gene expression in human chondrocytes.
abstract: BACKGROUND
  Conventional treatments for the articular diseases are often effective for symptom relief, but can also cause significant side effects and do not slow the progression of the disease. Several natural substances have been shown to be effective at relieving the symptoms of osteoarthritis (OA), and preliminary evidence suggests that some of these compounds may exert a favorable influence on the course of the disease. The objective of this study was to investigate the anti-inflammatory/chondroprotective potential of a Herbal and amino acid mixture containing extract of the Uncaria tomentosa, Boswellia spp., Lepidium meyenii and L-Leucine on the IL-1β-induced production of nitric oxide (NO), glycosaminoglycan (GAG), matrix metalloproteinases (MMPs), aggrecan (ACAN) and type II collagen (COL2A1) in human OA chondrocytes and OA cartilage explants.
  
  
  METHODS
  Primary OA chondrocytes or OA cartilage explants were pretreated with Herbal-Leucine mixture (HLM, 1-10 μg/ml) and then stimulated with IL-1β (5 ng/ml). Effect of HLM on IL-1β-induced gene expression of iNOS, MMP-9, MMP-13, ACAN and COL2A1 was verified by real time-PCR. Estimation of NO and GAG release in culture supernatant was done using commercially available kits.
  
  
  RESULTS
  HLM tested in these in vitro studies was found to be an effective anti-inflammatory agent, as evidenced by strong inhibition of iNOS, MMP-9 and MMP-13 expression and NO production in IL-1β-stimulated OA chondrocytes (p < 0.05). Supporting these gene expression results, IL-1β-induced cartilage matrix breakdown, as evidenced by GAG release from cartilage explants, was also significantly blocked (p < 0.05). Moreover, in the presence of herbal-Leucine mixture (HLM) up-regulation of ACAN and COL2A1 expression in IL-1β-stimulated OA chondrocytes was also noted (p < 0.05). The inhibitory effects of HLM were mediated by inhibiting the activation of nuclear factor (NF)-kB in human OA chondrocytes in presence of IL-1β.
  
  
  CONCLUSIONS
  Our data suggests that HLM could be chondroprotective and anti-inflammatory agent in arthritis, switching chondrocyte gene expression from catabolic direction towards anabolic and regenerative, and consequently this approach may be potentially useful as a new adjunct therapeutic/preventive agent for OA or injury recovery.
authors.affiliation: Department of Medicine/Rheumatology, MetroHealth Medical Center, Case Western Reserve University, Cleveland, Ohio 44109, USA.
authors.forename: Nahid
authors.initials: N
authors.lastname: Akhtar
authors.forename: Mark Js
authors.initials: MJ
authors.lastname: Miller
authors.forename: Tariq M
authors.initials: TM
authors.lastname: Haqqi
mesh_terms: D029743:Boswellia / Q000737:chemistry; D002358:Cartilage, Articular / Q000187:drug effects / Q000276:immunology*; D020948:Cat's Claw / Q000737:chemistry; D002478:Cells, Cultured; D019902:Chondrocytes / Q000187:drug effects / Q000276:immunology*; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D053583:Interleukin-1beta / Q000009:adverse effects* / Q000276:immunology; D029686:Lepidium / Q000737:chemistry; D007930:Leucine / Q000494:pharmacology*; D008297:Male; D008875:Middle Aged; D010003:Osteoarthritis / Q000139:chemically induced / Q000188:drug therapy / Q000235:genetics* / Q000276:immunology; D010936:Plant Extracts / Q000494:pharmacology*
chemical_list: D053583:Interleukin-1beta; D010936:Plant Extracts; D007930:Leucine