Expression and activity of hydrogen sulfide generating enzymes in murine macrophages stimulated with lipopolysaccharide and interferon-γ

Abstract

Murine macrophages of the J774A.1 line are hydrogen sulphide-producing cells with the primary role of γ-cystathionase (CTH) and secondary role of 3-mercaptopyruvate sulfurtransferase (limited by cysteine availability) and with a negligible role of cystathionine β-synthase (CBS) in H 2 S generation. J774A.1 cells stimulation with lipopolysaccharide (LPS) or interferongamma (IFNγ) resulted in decreased H 2 S levels after 24 h of incubation; however, they were restored to the control level after 48 h. Negligible CBS expression and activity in J774A.1 cells can result in homocysteine availability for CTH-catalyzed, H 2 S-generating reactions. This was supported by an increased CTH expression (IFNγ, 24 h and 48 h, and LPS, 48 h) and activity (24 h, LPS) in the stimulated cells. The results confirm the suggested feedback regulation between CBS and CTH.

The role of hydrogen sulphide signalling in macrophage activation

“…A decrease of H 2 S level in murine macrophages following 24 hr of LPS or IFN-c stimulation was observed (early phase), but the H 2 S content was restored to normal level after 48 hr of stimulation (late phase), which was associated with the feedback regulation between CBS and CSE. 54 It is worthy of note that H 2 S production was correlated with LPS-induced macrophage late-stage apoptosis, which could be blocked by the addition of H 2 S inhibitor. 55 Therefore, it is possible that sustained LPS stimulation renders macrophages that undergo apoptosis through the production of H 2 S (Fig.…”

“…In line with above observations, a time‐dependent change of H 2 S content in macrophages was found following activation. A decrease of H 2 S level in murine macrophages following 24 hr of LPS or IFN‐γ stimulation was observed (early phase), but the H 2 S content was restored to normal level after 48 hr of stimulation (late phase), which was associated with the feedback regulation between CBS and CSE 54 . It is worthy of note that H 2 S production was correlated with LPS‐induced macrophage late‐stage apoptosis, which could be blocked by the addition of H 2 S inhibitor 55 .…”

H2S, Polysulfides, and Enzymes: Physiological and Pathological Aspects

“…Thus, an argument may be put forward about homeostasis of sulfane sulfur levels in biological systems. Moreover, a negative feedback regulation between CBS and CTH was suggested by Nandi and Mishra [39] and confirmed by Bronowicka et al [38]. The adaptive cellular response to stimulation with both IFNγ and LPS caused a decreased level of H 2 S-associated low CBS expression and increased CTH expression.…”

“…To understand the physiological function of sulfane sulfur, its levels in biological samples (tissues, cell cultures) were determined using the reaction with cyanide, and subsequently investigating the thiocyanate yield of the complex with Fe 3+ , which is detectable by spectrophotometry [35]. Although the method is not very sensitive, it showed that sulfane sulfur levels were quite similar in various animal tissues [36,37] and in murine macrophages despite stimulation with lipopolysaccharide and interferon-γ [38]. Thus, an argument may be put forward about homeostasis of sulfane sulfur levels in biological systems.…”

Cystathionine γ-lyase and hydrogen sulfide modulates glucose transporter Glut1 expression via NF-κB and PI3k/Akt in macrophages during inflammation

“…CSE expression in macrophages is well known to play a significant role in H 2 S generation during the immune response [ 7 – 9 ]. However, the role of CBS for the generation of H 2 S in macrophages is reported to be negligible during an inflammatory response and therefore was not investigated here [ 38 – 40 ]. CSE expression in macrophages highlights these systems’ importance in macrophage biological activities.…”