Effect of hydrogen sulfide sources on inflammation and catabolic markers on interleukin 1β-stimulated human articular chondrocytes.
OBJECTIVE
Hydrogen sulfide (H2S), the third gasotransmitter together with NO and CO, is emerging as a regulator of inflammation. To test if it might offer therapeutic value in the treatment of osteoarthritis (OA) we evaluated the effects of two exogenous sources of H2S, NaSH and GYY4137, on inflammation and catabolic markers that characterize OA.


METHODS
Human chondrocytes (CHs) were isolated from OA tissue. Cells were stimulated with a pro-inflammatory cytokine (interleukin-1β, IL1β, 5 ng/ml) and the ability of the two H2S sources to ameliorate its effects on the cells was tested. Nitric oxide (NO) production was quantified through the Griess reaction. Protein levels of inducible NO synthase (NOS2) and matrix metalloproteinase 13 (MMP13) were visualized through immunocytochemistry (ICC). Relative mRNA expression was quantified with qRT-PCR. Prostaglandin-2 (PGE-2), interleukin 6 (IL6) and MMP13 levels were measured with specific EIAs. NFκB nuclear translocation was visualized with immunofluorescence.


RESULTS
Both H2S sources led to significant reductions in NO, PGE-2, IL6 and MMP13 released by the cells and at the protein level. This was achieved by downregulation of relevant genes involved in the synthesis routes of these molecules, namely NOS2, cyclooxigenase-2 (COX2), prostaglandin E synthase (PTGES), IL6 and MMP13. NFκB nuclear translocation was also reduced.


CONCLUSIONS
NaSH and GYY4137 show anti-inflammatory and anti-catabolic properties when added to IL1β activated osteoarthritic CHs. Supplementation with exogenous H2S sources can regulate the expression of relevant genes in OA pathogenesis and progression, counteracting IL1β pro-inflammatory signals that lead to cartilage destruction in part by reducing NFκB activation.
CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), Spain; Grupo de Bioingieneria Tisular y Terapia Celular (CBTTC), Servicio de Reumatología, Instituto de, Investigacion Biomédica (INIBIC), Complexo Hospitalario Universitario A Coruna (CHUAC), A Coruña, Spain. Electronic address: Elena.fernandez.burguera@sergas.es.
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EF
Burguera
CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), Spain; Departmento de Medicina, INIBIC-Universidad de A Coruña, A Coruña, Spain. Electronic address: angela.vela.anero@sergas.es.
A
A
Vela-Anero
CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), Spain; Grupo de Bioingieneria Tisular y Terapia Celular (CBTTC), Servicio de Reumatología, Instituto de, Investigacion Biomédica (INIBIC), Complexo Hospitalario Universitario A Coruna (CHUAC), A Coruña, Spain. Electronic address: joana.cristina.silva.magalhaes@sergas.es.
J
J
Magalhães
Departmento de Medicina, INIBIC-Universidad de A Coruña, A Coruña, Spain. Electronic address: rmf@udc.es.
R
R
Meijide-Faílde
CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), Spain; Grupo de Bioingieneria Tisular y Terapia Celular (CBTTC), Servicio de Reumatología, Instituto de, Investigacion Biomédica (INIBIC), Complexo Hospitalario Universitario A Coruna (CHUAC), A Coruña, Spain. Electronic address: fblagar@sergas.es.
F J
FJ
Blanco
D000368:Aged; D000369:Aged, 80 and over; D015415:Biomarkers / Q000378:metabolism; D002358:Cartilage, Articular / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D002478:Cells, Cultured; D019902:Chondrocytes / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D015232:Dinoprostone / Q000378:metabolism; D005260:Female; D006801:Humans; D006862:Hydrogen Sulfide / Q000378:metabolism*; D007249:Inflammation / Q000378:metabolism / Q000473:pathology*; D053583:Interleukin-1beta / Q000494:pharmacology*; D015850:Interleukin-6 / Q000378:metabolism; D008297:Male; D053509:Matrix Metalloproteinase 13 / Q000378:metabolism; D008660:Metabolism / Q000187:drug effects*; D009025:Morpholines / Q000494:pharmacology; D016328:NF-kappa B / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D019001:Nitric Oxide Synthase / Q000378:metabolism; D009946:Organothiophosphorus Compounds / Q000494:pharmacology; D010003:Osteoarthritis / Q000378:metabolism / Q000473:pathology*; D013440:Sulfides / Q000494:pharmacology*
D015415:Biomarkers; C529376:GYY 4137; D053583:Interleukin-1beta; D015850:Interleukin-6; D009025:Morpholines; D016328:NF-kappa B; D009946:Organothiophosphorus Compounds; D013440:Sulfides; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D053509:Matrix Metalloproteinase 13; C025451:sodium bisulfide; D015232:Dinoprostone; D006862:Hydrogen Sulfide
Human articular chondrocytes; Hydrogen sulfide; Inflammation; Metalloproteinase; Osteoarthritis