Chondroitin sulphate inhibits NF-κB activity induced by interaction of pathogenic and damage associated molecules.
To evaluate the anti-inflammatory mechanism of action of Chondroitin Sulphate (CS).



THP-1 macrophages were cultured with a range of sizes and concentrations of HA fragments with TLR4 (LPS in a physiologically relevant concentration determined by analyses of sera of a community clinic ascertained knee osteoarthritis (OA) cohort) or TLR2 (heat killed listeria bacteria) agonists and varying concentrations of CS in a physiologically relevant range (10-200 μg/ml). We measured IL-1β release, intracellular IL-1β, proIL-1β, caspase-1 and NF-κB activity and DNA binding activity of NF-κB transcription factors from nuclear and cytoplasmic extracts.



Serum LPS was significantly associated with radiographic knee joint space narrowing (JSN) (P = 0.02) in the OA cohort (n = 40). The priming dose of LPS used for these experiments (10 ng/ml) was below the lowest serum concentration of the OA cohort (median 47.09, range 14.43-81.36 ng/ml). Priming doses of LPS and HA fragments alone did not elicit an inflammatory response. However, primed with LPS, HA fragments produced large dose-dependent increases in IL-1β that were inhibitable by CS. CS did not inhibit caspase-1 activity but in physiologically achievable concentrations, attenuated NF-κB activity induced by either the TLR4 (LPS 1000 ng/ml) or TLR2 agonists alone or in combination with HA fragments. LPS induced and CS significantly reduced activity of canonical NF-κB transcription factors, p65, p50, c-Rel and RelB.



Subinflammatory concentrations of pathogenic (LPS, listeria) and damage associated (HA) molecules interact to induce macrophage-related inflammation. CS works upstream of the inflammasome by inhibiting activation of NF-κB transcription factors.
Duke Molecular Physiology Institute, Duke University School of Medicine, Durham, USA. Electronic address: stablert@hotmail.com.
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Stabler
Duke Molecular Physiology Institute, Duke University School of Medicine, Durham, USA; Department of Orthopedic Surgery, West China Hospital, West China Medical School, SiChuan University, ChengDu, SiChuan Province, People's Republic of China. Electronic address: Zey.huang@gmail.com.
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Huang
Pre-Clinical R&D Area, Bioibérica, S. A., Barcelona, Spain. Electronic address: lmontell@bioiberica.com.
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Montell
Pre-Clinical R&D Area, Bioibérica, S. A., Barcelona, Spain. Electronic address: jverges@bioiberica.com.
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Vergés
Duke Molecular Physiology Institute, Duke University School of Medicine, Durham, USA; Division of Rheumatology, Duke University School of Medicine, Durham, USA. Electronic address: kraus004@duke.edu.
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Kraus
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D002809:Chondroitin Sulfates / Q000494:pharmacology*; D005260:Female; D006801:Humans; D053583:Interleukin-1beta / Q000378:metabolism; D008070:Lipopolysaccharides / Q000097:blood; D008297:Male; D008875:Middle Aged; D016328:NF-kappa B / Q000037:antagonists & inhibitors*; D020370:Osteoarthritis, Knee / Q000378:metabolism; D000074084:THP-1 Cells
D000893:Anti-Inflammatory Agents; D053583:Interleukin-1beta; D008070:Lipopolysaccharides; D016328:NF-kappa B; D002809:Chondroitin Sulfates
Chondroitin sulphate; Hyaluronan; Inflammation; Macrophages; Osteoarthritis