In vitro study of intracellular IL-1beta production and beta1 integrins expression in stimulated chondrocytes--effect of rhein.
The purpose of the present study was to investigate the intracellular IL-1beta production and beta1 integrins (alpha4/beta1 and alpha5/beta1) expression on chondrocytes. Chondroytes monolayer (human chondrosarcoma cell line HEM-C55) were incubated for 12, 24 and 48 hours in the presence of Tumor Necrosis Factor-alpha (TNF-alpha, Sigma, France) or recombinant human IL-1alpha (rh-IL1alpha, Becton Dickinson, France). After direct immunolabelling, cells were either analyzed on FACScan flow cytometer (Becton Dickinson, France), or observed under an epi-fluorescence inverted microscope equipped with the CellScan EPR optical scanning acquisition system (IPLab-Scanalytics, USA). We found that the IL-1beta mean fluorescence intensity in flow cytometry and in 3D microscopy was increased in the presence of TNF-alpha or rh-IL-1alpha, and alpha4/beta1 or alpha5/beta1 expression was higher on stimulated cells than on control cells. On the other hand, we have evaluated the in vitro effects of rhein (10(-5) M, Negma, France), an active metabolite of diacerein, on the intracellular IL-1beta and beta1 integrins expressed by stimulated or no-stimulated chondrocytes. The results indicated that rhein leads to a reduction of IL-1beta synthesis whereas a weak decrease of beta1 integrins receptors expression is observed. From this study, it seems that rhein partially reduce cytokine-induced intracellular IL-1beta production, and it has a weak action on alpha4/beta1 or alpha5/beta1 receptors.
Gigant-Huselstein
C
C
Mécanique et Ingénierie Cellulaire et Tissulaire, LEMTA UMR INPL CNRS 7563 et IFR 111 Bioingénierie, Faculté de Médecine, 54505 Vandoeuvre-lès-Nancy, France.
Dumas
D
D
Payan
E
E
Muller
S
S
Bensoussan
D
D
Netter
P
P
Stoltz
J F
JF
D000880:Anthraquinones / Q000494:pharmacology; D002358:Cartilage, Articular / Q000276:immunology*; D019902:Chondrocytes / Q000276:immunology*; D002813:Chondrosarcoma; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005434:Flow Cytometry; D006801:Humans; D039041:Integrin alpha4beta1; D019012:Integrin beta1 / Q000096:biosynthesis*; D016023:Integrins / Q000378:metabolism; D007375:Interleukin-1 / Q000096:biosynthesis* / Q000494:pharmacology; D008856:Microscopy, Fluorescence; D017469:Receptors, Fibronectin / Q000378:metabolism; D017472:Receptors, Interleukin-1 / Q000378:metabolism; D016201:Receptors, Lymphocyte Homing / Q000378:metabolism; D011994:Recombinant Proteins / Q000494:pharmacology; D013268:Stimulation, Chemical; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D000880:Anthraquinones; D004791:Enzyme Inhibitors; D039041:Integrin alpha4beta1; D019012:Integrin beta1; D016023:Integrins; D007375:Interleukin-1; D017469:Receptors, Fibronectin; D017472:Receptors, Interleukin-1; D016201:Receptors, Lymphocyte Homing; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; C020491:rhein