Medical Hydrology and Balneology: Environmental Aspects

Hydrogen sulfide as an anti-inflamatory mediator
in osteoarthritis
F-Burguera E (1-2) , Vela A (3) , Blanco FJ (1-2) , Meijide R (3)
(1) Tissue Engineering and Cellular Therapy Group (CBBTC-CHUAC),
CIBER-BBN/ISCIII, Spain
(2) Reumatology service, INIBIC, University Hospital A Coruña, A Coruña, Spain
(3) Department of Medicine, INIBIC-A Coruña, University A Coruña, Spain
rmf@udc.es
Introduction and Objectives
Hydrogen sulfide (H2S) has in the past been associated with environmental toxicity;
however, balneotherapy has made use of sulfur waters as complementary
therapy in the treatment of rheumatic diseases such as osteoarthritis. Hydrogen
sulfide, the principal sulfur-containing compound of these medicinal waters has
been recently proposed as an endogenous mediator of inflammation and an antioxidant
in osteoarthritis (OA). The aim of this work was to study the effects of different
concentrations of two sulfur releasing compounds on human articular chondrocytes
from osteoarthritic (OA) tissue.
Materials and Methods
We analyzed the effects of the addition of different concentrations of either a
fast or slow release H2S donor (NaHS or GYY4137, respectively) on the levels of
nitric oxide (NO) production with Griess reagent and the expression of iNOS protein
through inmunocytochemistry (ICC). We also measured the effects on the
production of reactive oxygen species (ROS) by inmunofluorescence with the use
of dihydrorhodamine 123, and the expression of superoxide dismutase 2 (SOD2)
mRNA with qRT-PCR. For all studies cells were seeded with a known density and
kept in 5% CO2 in a humidified atmosphere at 37ºC for 48 hours. Prior to the
determinations cells were stimulated with 5 ng/ml of IL-1β and the different concentrations
of NaHS and GYY4137 (ranging from 50 µM to 1000 µM).
Results
None of the concentrations of hydrogen sulfide donors used were seen to cause
cell death. Stimulation of cells with IL-1β caused an increase in the production of
NO up to about 120 mM when compared with unstimulated cells (approximately 5
mM NO). Addition of GYY4137 led to a concentration-dependent reduction of up
to 33 mM for 1000 µM, whereas for NaHS the reduction was limited to 96 mM
with the highest concentration (1000 µM). When quantifying iNOS expression with
ICC a 2.5% positivity was detected in the basal conditions; this increased up to 9%
when cells were stimulated with IL-1β. Addition of the S-releasing compounds led
Balnea
2012, núm. 6, 200-201
200
ISBN: 978-84-669-1887-0
978-84-669-3482-4