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.
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.
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.
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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The datasets used and/or analysed during the current study are available from the corresponding author on reasonable request.
Munford RS. Endotoxemia-menace, marker, or mistake? J Leukoc Biol. 2016;100(4):687–98. https://doi.org/10.1189/jlb.3RU0316-151R.
Martins IJ. Overnutrition determines LPS regulation of mycotoxin induced neurotoxicity in neurodegenerative diseases. Int J Mol Sci. 2015;16(12):29554–73. https://doi.org/10.3390/ijms161226190.
Wang YW, Zhou Q, Zhang X, Qian QQ, Xu JW, Ni PF, et al. Mild endoplasmic reticulum stress ameliorates lipopolysaccharide-induced neuroinflammation and cognitive impairment via regulation of microglial polarization. J Neuroinflamm. 2017;14(1):233. https://doi.org/10.1186/s12974-017-1002-7.
Kimura H. Physiological roles of hydrogen sulfide and polysulfides. In: Moore P, Whiteman M, editors. Chemistry, biochemistry and pharmacology of hydrogen sulfide. Handbook of experimental pharmacology, vol. 230. Springer, Cham;2015. p. 61–81. https://doi.org/10.1007/978-3-319-18144-8_3.
Rose P, Dymock BW, Moore PK. GYY4137, a novel water-soluble, H2S-releasing molecule. Methods Enzymol. 2015;554:143–67. https://doi.org/10.1016/bs.mie.2014.11.014.
Whiteman M, Li L, Rose P, Tan CH, Parkinson DB, Moore PK. The effect of hydrogen sulfide donors on lipopolysaccharide-induced formation of inflammatory mediators in macrophages. Antioxid Redox Signal. 2010;12(10):1147–54. https://doi.org/10.1089/ars.2009.2899.
Li L, Fox B, Keeble J, Salto-Tellez M, Winyard PG, Wood ME, et al. The complex effects of the slow-releasing hydrogen sulfide donor GYY4137 in a model of acute joint inflammation and in human cartilage cells. J Cell Mol Med. 2013;17(3):365–76. https://doi.org/10.1111/jcmm.12016.
Kaya-Yasar Y, Karaman Y, Bozkurt TE, Onder SC, Sahin-Erdemli I. Effects of intranasal treatment with slow (GYY4137) and rapid (NaHS) donors of hydrogen sulfide in lipopolysaccharide-induced airway inflammation in mice. Pulm Pharmacol Ther. 2017;45:170–80. https://doi.org/10.1016/j.pupt.2017.06.006.
Meng G, Wang J, Xiao Y, Bai W, Xie L, Shan L, et al. GYY4137 protects against myocardial ischemia and reperfusion injury by attenuating oxidative stress and apoptosis in rats. J Biomed Res. 2015;29(3):203–13. https://doi.org/10.7555/jbr.28.20140037.
Bobkova NV, Garbuz DG, Nesterova I, Medvinskaya N, Samokhin A, Alexandrova I, et al. Therapeutic effect of exogenous hsp70 in mouse models of Alzheimer’s disease. J Alzheimer’s Dis. 2014;38(2):425–35. https://doi.org/10.3233/jad-130779.
Bobkova NV, Evgen'ev M, Garbuz DG, Kulikov AM, Morozov A, Samokhin A, et al. Exogenous Hsp70 delays senescence and improves cognitive function in aging mice. Proc Natl Acad Sci USA. 2015;112(52):16006–11. https://doi.org/10.1073/pnas.1516131112.
Lu Z, Zhao T, Tao L, Yu Q, Yang Y, Cheng J, et al. Cystathionine beta-synthase-derived hydrogen sulfide correlates with successful aging in mice. Rejuvenation Res. 2019. https://doi.org/10.1089/rej.2018.2166.
Gerasimova E, Lebedeva J, Yakovlev A, Zefirov A, Giniatullin R, Sitdikova G. Mechanisms of hydrogen sulfide (H2S) action on synaptic transmission at the mouse neuromuscular junction. Neuroscience. 2015;303:577–85. https://doi.org/10.1016/j.neuroscience.2015.07.036.
Lakshmikanth CL, Jacob SP, Chaithra VH, de Castro-Faria-Neto HC, Marathe GK. Sepsis: in search of cure. Inflamm Res. 2016;65(8):587–602. https://doi.org/10.1007/s00011-016-0937-y.
Petrushanko IY, Melnikova EV, Yurinskaya MM, Vinokurov MG, Suslikov AV, Mitkevich VA, et al. Influence of the donor of hydrogen sulfide GYY4137 on the activation of human neutrophils by E. coli lipopolysaccharides. Mol Biol. 2019;53(1):101–8. https://doi.org/10.1134/s0026898419010130.
Rozhkova E, Yurinskaya M, Zatsepina O, Garbuz D, Karpov V, Surkov S, et al. Exogenous mammalian extracellular HSP70 reduces endotoxin manifestations at the cellular and organism levels. Ann N Y Acad Sci. 2010;1197:94–107. https://doi.org/10.1111/j.1749-6632.2009.05375.x.
Yurinskaya MM, Kochetkova OY, Shabarchina LI, Antonova OY, Suslikov AV, Evgen'ev MB, et al. Encapsulated Hsp70 decreases endotoxin-induced production of ROS and TNFalpha in human phagocytes. Cell Stress Chaperones. 2017;22(1):163–71. https://doi.org/10.1007/s12192-016-0743-z.
Yurinskaya MM, Mit'kevich VA, Evgen'ev MB, Makarov AA, Vinokurov MG. Heat-shock protein HSP70 reduces the secretion of TNFalpha by neuroblastoma cells and human monocytes induced with beta-amyloid peptides. Mol Biol. 2016;50(6):1053–6. https://doi.org/10.7868/s0026898416060239.
Bolger AM, Lohse M, Usadel B. Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics. 2014;30(15):2114–200. https://doi.org/10.1093/bioinformatics/btu170.
Dobin A, Davis CA, Schlesinger F, Drenkow J, Zaleski C, Jha S, et al. STAR: ultrafast universal RNA-seq aligner. Bioinformatics. 2013;29(1):15–211. https://doi.org/10.1093/bioinformatics/bts635.
Liao Y, Smyth GK, Shi W. FeatureCounts: an efficient general purpose program for assigning sequence reads to genomic features. Bioinformatics. 2014;30(7):923–30. https://doi.org/10.1093/bioinformatics/btt656.
Wang L, Wang S, Li W. RSeQC: quality control of RNA-seq experiments. Bioinformatics. 2012;28(16):2184–5. https://doi.org/10.1093/bioinformatics/bts356.
Krasnov GS, Dmitriev AA, Kudryavtseva AV, Shargunov AV, Karpov DS, Uroshlev LA, et al. PPLine: an automated pipeline for SNP, SAP, and splice variant detection in the context of proteogenomics. J Proteom Res. 2015;14(9):3729–37. https://doi.org/10.1021/acs.jproteome.5b00490.
Robinson MD, McCarthy DJ, Smyth GK. edgeR: a Bioconductor package for differential expression analysis of digital gene expression data. Bioinformatics. 2010;26(1):139–40. https://doi.org/10.1093/bioinformatics/btp616.
Yu G, Wang LG, Han Y, He QY. clusterProfiler: an R package for comparing biological themes among gene clusters. OMICS. 2012;16(5):284–7. https://doi.org/10.1089/omi.2011.0118.
Ferrington DA, Gregerson DS. Immunoproteasomes: structure, function, and antigen presentation. Prog Mol Biol Transl Sci. 2012;109:75–112. https://doi.org/10.1016/b978-0-12-397863-9.00003-1.
Pan W, Kang Y. Gut microbiota and chronic kidney disease: implications for novel mechanistic insights and therapeutic strategies. Int Urol Nephrol. 2018;50(2):289–99. https://doi.org/10.1007/s11255-017-1689-5.
Kuzmich NN, Sivak KV, Chubarev VN, Porozov YB, Savateeva-Lyubimova TN, Peri F. TLR4 signaling pathway modulators as potential therapeutics in inflammation and sepsis. Vaccines. 2017;5(4):34–75. https://doi.org/10.3390/vaccines5040034.
Shapouri-Moghaddam A, Mohammadian S, Vazini H, Taghadosi M, Esmaeili SA, Mardani F, et al. Macrophage plasticity, polarization, and function in health and disease. J Cell Physiol. 2018;233(9):6425–40. https://doi.org/10.1002/jcp.26429.
Chen LS, Singh SP, Schuster M, Grinenko T, Bornstein SR, Kanczkowski W. RNA-seq analysis of LPS-induced transcriptional changes and its possible implications for the adrenal gland dysregulation during sepsis. J Steroid Biochem Mol Biol. 2019;191:105360. https://doi.org/10.1016/j.jsbmb.2019.04.009.
Kim SJ, Kim HM. Dynamic lipopolysaccharide transfer cascade to TLR4/MD2 complex via LBP and CD14. BMB Rep. 2017;50(2):55–7. https://doi.org/10.5483/bmbrep.2017.50.2.011.
Pfeiffer JR, McAvoy BL, Fecteau RE, Deleault KM, Brooks SA. CARHSP1 is required for effective tumor necrosis factor alpha mRNA stabilization and localizes to processing bodies and exosomes. Mol Cell Biol. 2011;31(2):277–86. https://doi.org/10.1128/mcb.00775-10.
Mouzaoui S, Rahim I, Djerdjouri B. Aminoguanidine and curcumin attenuated tumor necrosis factor (TNF)-alpha-induced oxidative stress, colitis and hepatotoxicity in mice. Int Immunopharmacol. 2012;12(1):302–11. https://doi.org/10.1016/j.intimp.2011.10.010.
Blaser H, Dostert C, Mak TW, Brenner D. TNF and ROS crosstalk in inflammation. Trends Cell Biol. 2016;26(4):249–61. https://doi.org/10.1016/j.tcb.2015.12.002.
Sakuma S, Minamino S, Takase M, Ishiyama Y, Hosokura H, Kohda T, et al. Hydrogen sulfide donor GYY4137 suppresses proliferation of human colorectal cancer Caco-2 cells by inducing both cell cycle arrest and cell death. Heliyon. 2019;5(8):e02244. https://doi.org/10.1016/j.heliyon.2019.e02244.
Dauphinee SM, Karsan A. Lipopolysaccharide signaling in endothelial cells. Lab Investig. 2006;86(1):9–22. https://doi.org/10.1038/labinvest.3700366.
Reis J, Guan XQ, Kisselev AF, Papasian CJ, Qureshi AA, Morrison DC, et al. LPS-induced formation of immunoproteasomes: TNF-alpha and nitric oxide production are regulated by altered composition of proteasome-active sites. Cell Biochem Biophys. 2011;60(1–2):77–88. https://doi.org/10.1007/s12013-011-9182-8.
Shen J, Reis J, Morrison DC, Papasian C, Raghavakaimal S, Kolbert C, et al. Key inflammatory signaling pathways are regulated by the proteasome. Shock. 2006;25(5):472–84. https://doi.org/10.1097/01.shk.0000209554.46704.64.
Kustanova GA, Murashev AN, Karpov VL, Margulis BA, Guzhova IV, Prokhorenko IR, et al. Exogenous heat shock protein 70 mediates sepsis manifestations and decreases the mortality rate in rats. Cell Stress Chaperones. 2006;11(3):276–86. https://doi.org/10.1379/csc-195r.1.
Yurinskaya MM, Mitkevich VA, Kozin SA, Evgen'ev MB, Makarov AA, Vinokurov MG. HSP70 protects human neuroblastoma cells from apoptosis and oxidative stress induced by amyloid peptide isoAsp7-Abeta(1–42). Cell Death Dis. 2015;6:e1977. https://doi.org/10.1038/cddis.2015.336.
Evgen'ev MB, Krasnov GS, Nesterova IV, Garbuz DG, Karpov VL, Morozov AV, et al. Molecular mechanisms underlying neuroprotective effect of intranasal administration of human Hsp70 in mouse model of Alzheimer’s disease. J Alzheimer’s Dis. 2017;59(4):1415–26. https://doi.org/10.3233/jad-170398.
Yadav V, Gao XH, Willard B, Hatzoglou M, Banerjee R, Kabil O. Hydrogen sulfide modulates eukaryotic translation initiation factor 2alpha (eIF2alpha) phosphorylation status in the integrated stress-response pathway. J Biol Chem. 2017;292(32):13143–53. https://doi.org/10.1074/jbc.M117.778654.
Aneja R, Odoms K, Dunsmore K, Shanley TP, Wong HR. Extracellular heat shock protein-70 induces endotoxin tolerance in THP-1 cells. J Immunol. 2006;177(10):7184–92. https://doi.org/10.4049/jimmunol.177.10.7184.
Asea A, Kraeft SK, Kurt-Jones EA, Stevenson MA, Chen LB, Finberg RW, et al. HSP70 stimulates cytokine production through a CD14-dependant pathway, demonstrating its dual role as a chaperone and cytokine. Nat Med. 2000;6(4):435–42. https://doi.org/10.1038/74697.
Yurinskaya M, Zatsepina OG, Vinokurov MG, Bobkova NV, Garbuz DG, Morozov AV, et al. The fate of exogenous human HSP70 introduced into animal cells by different means. Curr Drug Deliv. 2015;12(5):524–32.
Pockley AG, Henderson B. Extracellular cell stress (Heat shock) proteins—immune responses and disease: an overview. Philos Trans R Soc Lond Ser B Biol Sci. 2018;373(1738):20160522. https://doi.org/10.1098/rstb.2016.0522.
Filipovic MR, Zivanovic J, Alvarez B, Banerjee R. Chemical biology of H2S signaling through persulfidation. Chem Rev. 2018;118(3):1253–337. https://doi.org/10.1021/acs.chemrev.7b00205.
Hulina A, Grdic Rajkovic M, Jaksic Despot D, Jelic D, Dojder A, Cepelak I, et al. Extracellular Hsp70 induces inflammation and modulates LPS/LTA-stimulated inflammatory response in THP-1 cells. Cell Stress Chaperones. 2018;23(3):373–84. https://doi.org/10.1007/s12192-017-0847-0.
Lee KH, Jeong J, Yoo CG. Positive feedback regulation of heat shock protein 70 (Hsp70) is mediated through toll-like receptor 4-PI3K/Akt-glycogen synthase kinase-3beta pathway. Exp Cell Res. 2013;319(1):88–95. https://doi.org/10.1016/j.yexcr.2012.09.018.
Du M, Yuan L, Tan X, Huang D, Wang X, Zheng Z, et al. The LPS-inducible lncRNA Mirt2 is a negative regulator of inflammation. Nat Commun. 2017;8(1):2049. https://doi.org/10.1038/s41467-017-02229-1.
Ji K, Xue L, Cheng J, Bai Y. Preconditioning of H2S inhalation protects against cerebral ischemia/reperfusion injury by induction of HSP70 through PI3K/Akt/Nrf2 pathway. Brain Res Bull. 2016;121:68–74. https://doi.org/10.1016/j.brainresbull.2015.12.007.
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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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
- H2S donors
- LPS challenge
- Pro-inflammatory pathways
- Hsps genes