H2S counteracts proinflammatory effects of LPS through modulation of multiple pathways in human cells

Abstract

Background

Hydrogen sulfide donors reduce inflammatory signaling in vitro and in vivo. The biological effect mediated by H2S donors depends on the kinetics of the gas release from the donor molecule. However, the molecular mechanisms of H2S-induced immunomodulation were poorly addressed. Here, we studied the effect of two different hydrogen sulfide (H2S)-producing agents on the generation of the LPS-induced inflammatory mediators. Importantly, we investigated the transcriptomic changes that take place in human cells after the LPS challenge, combined with the pretreatment with a slow-releasing H2S donor-GYY4137.

Methods

We investigated the effects of GYY4137 and sodium hydrosulfide on the release of proinflammatory molecules such as ROS, NO and TNF-α from LPS-treated human SH-SY5Y neuroblastoma and the THP-1 promonocytic cell lines. Transcriptomic and RT-qPCR studies using THP-1 cells were performed to monitor the effects of the GYY4137 on multiple signaling pathways, including various immune-related and proinflammatory genes after combined action of LPS and GYY4137.

Results

The GYY4137 and sodium hydrosulfide differed in the ability to reduce the production of the LPS-evoked proinflammatory mediators. The pre-treatment with GYY4137 resulted in a drastic down-regulation of many TNF-α effectors that are induced by LPS treatment in THP-1 cells. Furthermore, GYY4137 pretreatment of LPS-exposed cells ameliorates the LPS-mediated induction of multiple pro-inflammatory genes and decreases expression of immunoproteasome genes. Besides, in these experiments we detected the up-regulation of several important pathways that are inhibited by LPS.

Conclusion

Based on the obtained results we believe that our transcriptomic analysis significantly contributes to the understanding of the molecular mechanisms of anti-inflammatory and cytoprotective activity of hydrogen sulfide donors, and highlights their potential against LPS challenges and other forms of inflammation.

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Availability of data and materials

The datasets used and/or analysed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

RNA sequencing was performed using the equipment of the Engelhardt Institute of Molecular Biology RAS “Genome” center (https://www.eimb.ru/rus/ckp/ccu_genome_c.php). This work has been supported by Grant of Russian Science Foundation No 17-74-30030 and Russian grant Program for Basic Science No 19-04-00109.

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MY and MV performed all experiments with both human cell cultures; GK and AR bioinformatic analysis of the transcriptome data; DK performed RNA isolation and RTPCR studies; OZ and LC isolated RNA and prepared libraries for transcriptomic analysis; SF prepared libraries and performed RTPCR studies; AM—Western blot analysis and proteasome studies, wrote the paper; ME—designed the research, wrote the paper. All authors read and approved the final manuscript.

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Correspondence to M. B. Evgen’ev.

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Yurinskaya, M.M., Krasnov, G.S., Kulikova, D.A. et al. H2S counteracts proinflammatory effects of LPS through modulation of multiple pathways in human cells. Inflamm. Res. 69, 481–495 (2020). https://doi.org/10.1007/s00011-020-01329-x

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Keywords

  • H2S donors
  • LPS challenge
  • Pro-inflammatory pathways
  • Hsps genes
  • Transcriptome