Inhibitors of p38 and ERK1/2 MAPkinase and hydrogen sulphide block constitutive and IL-1β-induced IL-6 and IL-8 expression in the human chondrocyte cell line C-28/I2

Abstract

Mitogen-activated protein kinases (MAPKs) play a central role in inflammatory processes, and their blockage represents pharmacological approaches in the treatment of autoimmune diseases like rheumatoid arthritis (RA). Alternatively, H(2)S has long been used in sulphur bath therapy for patients suffering from different types of rheumatic disorders, but reports about the beneficial effects of this form of therapy are controversial, rare and of poor scientific quality. The human chondrocyte cell line C-28/I2 was treated with two different MAPK inhibitors (SB203580 and U0126) or with various concentrations of the H(2)S donor Natrium hydrogen sulphide (NaHS). Thereafter, the secretion of IL-6 and IL-8 was quantified by enzyme-linked immunosorbent assays (ELISAs). The impact of NaHS on the regulation of p38 and ERK1/2 MAPK was confirmed by Western blot experiments. Furthermore, IL-6 and IL-8 expression was quantified by real-time polymerase chain reaction (RT-PCR) and ELISAs from cells which were exposed to SB203580, U0126 and NaHS and stimulated by IL-1β. The C-28/I2 cells constitutively expressed large quantities of IL-6 and IL-8. The data provided prove that in these cells, constitutive as well as IL-1β-induced IL-6 and IL-8 expression was partially and transiently blocked by the treatment of cells with both MAPK inhibitors and NaHS. Presented data seem to be important in evaluating the beneficial functions of MAPK inhibitors and H(2)S in immune-pathophysiological processes.

Periodic Mechanical Stress Stimulates the FAK Mitogenic Signal in Rat Chondrocytes Through ERK1/2 Activity

“…qPCR analysis was performed with the LightCycler System (Roche Diagnostics) using SYBR Premix Ex Taq II, as described previously [22]. The reaction was performed in a 20 µL mixture containing 2 µL cDNA.…”

Hydrogen Sulfide in Biochemistry and Medicine

“…H 2 S has been shown to downregulate several pro-inflammatory cytokines including NF-jB, TNF-a, IL-1b, IL-6, and IL-8 (55,98,144,151), to modulate leukocyte adhesion and leukocyte-mediated inflammation (55,181), to mediate the cardioprotection induced by ischemic postconditioning (241) and to protect from NF-jB and TNF-a mediated endotoxic shock (113). The powerful reducing/antioxidant/free radical scavenging properties of H 2 S can explain its wide-ranging anti-inflammatory and cytoprotective effects, including protection against: ischemiareperfusion injury in heart, brain, retina, liver, and intestine; endothelial dysfunction; hydrogen peroxide-induced damage in rat gastric epithelial cells; hyperhomocysteinemia in rats; methionine-and homocysteine-induced oxidative stress; and hemin-mediated oxidation of low-density lipoprotein (112).…”

Periodic Mechanical Stress Activates Integrin�1-Dependent Src-Dependent PLC?1-Independent Rac1 Mitogenic Signal in Rat Chondrocytes through ERK1/2

“…qPCR analysis was performed with the LightCycler System (Roche Diagnostics) using SYBR Premix Ex Taq II as previously described [28]. The reaction was performed in a 20-µL mixture containing 2 µL of the above cDNA.…”