Sulfur compounds block MCP-1 production by Mycoplasma fermentans-infected macrophages through NF-κB inhibition

J Transl Med. 2014 May 24:12:145. doi: 10.1186/1479-5876-12-145.

Abstract

Background and aims: Hydrogen sulfide (H2S), together with nitric oxide (NO) and carbon monoxide (CO), belongs to a family of endogenous signaling mediators termed "gasotransmitters". Recent studies suggest that H2S modulates many cellular processes and it has been recognized to play a central role in inflammation, in the cardiovascular and nervous systems. By infecting monocytes/macrophages with Mycoplasma fermentans (M.F.), a well-known pro-inflammatory agent, we evaluated the effects of H2S.

Methods: M.F.-infected cells were analyzed by ELISA and real time RT-PCR to detect the M.F. effects on MCP-1 and on MMP-12 expression. The role of two different H2S donors (NaHS and GYY4137) on MF-infected cells was determined by treating infected cells with H2S and then testing the culture supernatants for MCP-1 and on MMP-12 production by ELISA assay. In order to identify the pathway/s mediating H2S- anti-inflammatory activity, cells were also treated with specific pharmaceutical inhibitors. Cytoplasmic and nuclear accumulation of NF-κB heterodimers was analyzed.

Results: We show that H2S was able to reduce the production of pro-inflammatory cytokine MCP-1, that was induced in monocytes/macrophages during M.F. infection. Moreover, MCP-1 was induced by M.F. through Toll-like receptor (TLR)-mediated nuclear factor-κB (NF-κB) activation, as demonstrated by the fact that TLR inhibitors TIRAP and MyD88 and NF-κB inhibitor IKK were able to block the cytokine production. In contrast H2S treatment of M.F. infected macrophages reduced nuclear accumulation of NF-κB heterodimer p65/p52.

Conclusions: Our data demonstrate that under the present conditions H2S is effective in reducing Mycoplasma-induced inflammation by targeting the NF-κB pathway. This supports further studies for possible clinical applications.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Chemokine CCL2 / biosynthesis*
  • DNA Primers
  • Enzyme Induction
  • Enzyme-Linked Immunosorbent Assay
  • Humans
  • Hydrogen Sulfide / pharmacology*
  • Macrophages / microbiology*
  • Matrix Metalloproteinase 12 / biosynthesis
  • Mycoplasma fermentans / drug effects*
  • Mycoplasma fermentans / metabolism
  • NF-kappa B / antagonists & inhibitors*
  • Real-Time Polymerase Chain Reaction
  • U937 Cells

Substances

  • Chemokine CCL2
  • DNA Primers
  • NF-kappa B
  • Matrix Metalloproteinase 12
  • Hydrogen Sulfide