TY - JOUR
T1 - Monosodium urate crystals induce oxidative stress in human synoviocytes
AU - Zamudio-Cuevas, Yessica
AU - Martínez-Flores, Karina
AU - Fernández-Torres, Javier
AU - Loissell-Baltazar, Yahir A.
AU - Medina-Luna, Daniel
AU - López-Macay, Ambar
AU - Camacho-Galindo, Javier
AU - Hernández-Díaz, Cristina
AU - Santamaría-Olmedo, Mónica G.
AU - López-Villegas, Edgar Oliver
AU - Oliviero, Francesca
AU - Scanu, Anna
AU - Cerna-Cortés, Jorge Francisco
AU - Gutierrez, Marwin
AU - Pineda, Carlos
AU - López-Reyes, Alberto
N1 - Publisher Copyright:
© 2016 Zamudio-Cuevas et al.
PY - 2016/5/21
Y1 - 2016/5/21
N2 - Background: Gout is the most common inflammatory arthropathy of metabolic origin and it is characterized by intense inflammation, the underlying mechanisms of which are unknown. The aim of this study was to evaluate the oxidative stress in human fibroblast-like synoviocytes (FLS) exposed to monosodium urate (MSU) crystals, which trigger an inflammatory process. Methods: Human FLS isolated from synovial tissue explants were stimulated with MSU crystals (75 μg/mL) for 24 h. Cellular viability was evaluated by crystal violet staining, apoptosis was assessed using Annexin V, and the cellular content of reactive oxygen species (ROS) and nitrogen species (RNS) (O2 -, H2O2, NO) was assessed with image-based cytometry and fluorometric methods. In order to determine protein oxidation levels, protein carbonyls were detected through oxyblot analysis, and cell ultrastructural changes were assessed by transmission electron microscopy. Results: The viability of FLS exposed to MSU crystals decreased by 30% (P<0.05), while apoptosis increased by 42% (P=0.01). FLS stimulated with MSU crystals exhibited a 2.1-fold increase in H2O2 content and a 1.5-fold increase in O2 - and NO levels. Oxyblots revealed that the spots obtained from FLS protein lysates exposed to MSU crystals exhibited protein carbonyl immunoreactivity, which reflects the presence of oxidatively modified proteins. Concomitantly, MSU crystals triggered the induction of changes in the morphostructure of FLS, such as the thickening and discontinuity of the endoplasmic reticulum, and the formation of vacuoles and misfolded glycoproteins. Conclusions: Our results prove that MSU crystals induce the release of ROS and RNS in FLS, subsequently oxidizing proteins and altering the cellular oxidative state of the endoplasmic reticulum, which results in FLS apoptosis.
AB - Background: Gout is the most common inflammatory arthropathy of metabolic origin and it is characterized by intense inflammation, the underlying mechanisms of which are unknown. The aim of this study was to evaluate the oxidative stress in human fibroblast-like synoviocytes (FLS) exposed to monosodium urate (MSU) crystals, which trigger an inflammatory process. Methods: Human FLS isolated from synovial tissue explants were stimulated with MSU crystals (75 μg/mL) for 24 h. Cellular viability was evaluated by crystal violet staining, apoptosis was assessed using Annexin V, and the cellular content of reactive oxygen species (ROS) and nitrogen species (RNS) (O2 -, H2O2, NO) was assessed with image-based cytometry and fluorometric methods. In order to determine protein oxidation levels, protein carbonyls were detected through oxyblot analysis, and cell ultrastructural changes were assessed by transmission electron microscopy. Results: The viability of FLS exposed to MSU crystals decreased by 30% (P<0.05), while apoptosis increased by 42% (P=0.01). FLS stimulated with MSU crystals exhibited a 2.1-fold increase in H2O2 content and a 1.5-fold increase in O2 - and NO levels. Oxyblots revealed that the spots obtained from FLS protein lysates exposed to MSU crystals exhibited protein carbonyl immunoreactivity, which reflects the presence of oxidatively modified proteins. Concomitantly, MSU crystals triggered the induction of changes in the morphostructure of FLS, such as the thickening and discontinuity of the endoplasmic reticulum, and the formation of vacuoles and misfolded glycoproteins. Conclusions: Our results prove that MSU crystals induce the release of ROS and RNS in FLS, subsequently oxidizing proteins and altering the cellular oxidative state of the endoplasmic reticulum, which results in FLS apoptosis.
KW - Gout
KW - Monosodium urate crystals
KW - Oxidative stress
KW - Synoviocytes
UR - http://www.scopus.com/inward/record.url?scp=84969535035&partnerID=8YFLogxK
U2 - 10.1186/s13075-016-1012-3
DO - 10.1186/s13075-016-1012-3
M3 - Artículo
C2 - 27209322
SN - 1478-6354
VL - 18
JO - Arthritis Research and Therapy
JF - Arthritis Research and Therapy
IS - 1
M1 - 117
ER -