Abstract
Hydrogen sulfide (H2S) plays an important role in inflammation. We showed that macrophages expressed the H2S-forming enzyme cystathionine gamma-lyase (CSE) and produced H2S. Lipopolysaccharide (LPS) stimulated the CSE expression and H2S production rate. l-cysteine reduced LPS-induced nitric oxide (NO) production. CSE inhibitor blocked the inhibitory effect of l-cysteine. CSE knockdown increased, whereas CSE overexpression decreased LPS-induced NO production. Dexamethasone suppressed LPS-induced CSE expression and the H2S production rate as well as NO production. l-arginine increased, whereas NG-nitro-l-arginine methyl ester (l-NAME) decreased LPS-induced CSE expression and H2S production. Dexamethasone plus l-NAME significantly decreased LPS-induced CSE expression and H2S production compared to l-NAME. Our results suggest that macrophages are one of the H2S producing sources. H2S might exert anti-inflammatory effects by inhibiting NO production. Dexamethasone may directly inhibit CSE expression and H2S production, besides the NO-dependent way. Inhibition of H2S and NO production may be a mechanism by which glucocorticoids coordinate the balance between pro- and anti-inflammatory mediators during inflammation.
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Abbreviations
- H2S:
-
Hydrogen sulfide
- NO:
-
Nitric oxide
- CO:
-
Carbon monoxide
- CSE:
-
Cystathionine gamma-lyase
- CBS:
-
Cystathionine β-synthetase
- fMLP:
-
Formyl-methionyl-leucyl-phenylalanine
- TNF-α:
-
Tumor necrosis factor-α
- GCs:
-
Glucocorticoids
- GR:
-
Glucocorticoid receptor
- MIF:
-
Migration inhibitory factor
- LPS:
-
Lipopolysaccharide
- IL-1:
-
Interleukin-1
- IL-6:
-
Interleukin-6
- MTT:
-
3-[4, 5-Dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide
- PAG:
-
dl-propargylglycine
- l-NAME:
-
NG-nitro-l-arginine methyl ester
- iNOS:
-
Inducible nitric oxide synthase
- CD-FBS:
-
Charcoal-stripped FBS
- PLP:
-
Pyridoxal-5′-phosphate
- siRNA:
-
Interfering RNA
- RAW-EGFP-mCSE:
-
CSE-overexpression RAW264.7 cell lines
- RAW-mCSE siRNA:
-
CSE-knockdown RAW264.7 cell lines
- TBST:
-
Tris-buffered saline/Tween 20
References
Moore PK, Bhatia M, Moochhala S (2003) Hydrogen sulfide: from the smell of the past to the mediator of the future? Trends Pharmacol Sci 24:609–611
Wang R (2002) Two’s company, three’s a crowd: can H2S be the third endogenous gaseous transmitter? FASEB J 16:1792–1798
Bhatia M (2005) Hydrogen sulfide as a vasodilator. IUBMB Life 57:603–606
Qingyou Z, Junbao D, Weijin Z, Hui Y, Chaoshu T, Chunyu Z (2004) Impact of hydrogen sulfide on carbon monoxide/heme oxygenase pathway in the pathogenesis of hypoxic pulmonary hypertension. Biochem Biophys Res Commun 317:30–37
Zhao W, Zhang J, Lu Y, Wang R (2001) The vasorelaxant effect of H2S as a novel endogenous gaseous KATP channel opener. EMBO J 20:6008–6016
Li L, Bhatia M, Zhu YZ, Zhu YC, Ramnath RD, Wang ZJ, Anuar FB, Whiteman M, Salto-Tellez M, Moore PK (2005) Hydrogen sulfide is a novel mediator of lipopolysaccharide-induced inflammation in the mouse. Faseb J 19:1196–1198
Hui Y, Du J, Tang C, Bin G, Jiang H (2003) Changes in arterial hydrogen sulfide (H(2)S) content during septic shock and endotoxin shock in rats. J Infect 47:155–160
Zhi L, Ang AD, Zhang H, Moore PK, Bhatia M (2007) Hydrogen sulfide induces the synthesis of proinflammatory cytokines in human monocyte cell line U937 via the ERK-NF-kappaB pathway. J Leukoc Biol 81:1322–1332
Collin M, Anuar FB, Murch O, Bhatia M, Moore PK, Thiemermann C (2005) Inhibition of endogenous hydrogen sulfide formation reduces the organ injury caused by endotoxemia. Br J Pharmacol 146:498–505
Bhatia M, Wong FL, Fu D, Lau HY, Moochhala SM, Moore PK (2005) Role of hydrogen sulfide in acute pancreatitis and associated lung injury. Faseb J 19:623–625
Whiteman M, Armstrong JS, Chu SH, Jia-Ling S, Wong BS, Cheung NS, Halliwell B, Moore PK (2004) The novel neuromodulator hydrogen sulfide: an endogenous peroxynitrite ‘scavenger’? J Neurochem 90:765–768
Fiorucci S, Antonelli E, Distrutti E, Rizzo G, Mencarelli A, Orlandi S, Zanardo R, Renga B, Di Sante M, Morelli A, Cirino G, Wallace JL (2005) Inhibition of hydrogen sulfide generation contributes to gastric injury caused by anti-inflammatory nonsteroidal drugs. Gastroenterology 129:1210–1224
Zanardo RC, Brancaleone V, Distrutti E, Fiorucci S, Cirino G, Wallace JL (2006) Hydrogen sulfide is an endogenous modulator of leukocyte-mediated inflammation. FASEB J 20:2118–2120
Mariggio MA, Pettini F, Fumarulo R (1997) Sulfide influence onpolymorphonuclear functions: a possible role for Ca2+ involvement. Immunopharmacol Immunotoxicol 19:393–404
Mariggio MA, Minunno V, Riccardi S, Santacroce R, De Rinaldis P, Fumarulo R (1998) Sulfide enhancement of PMN apoptosis. Immunopharmacol Immunotoxicol 20:399–408
Hu LF, Wong PT, Moore PK, Bian JS (2007) Hydrogen sulfide attenuates lipopolysaccharide-induced inflammation by inhibition of p38 mitogen-activated protein kinase in microglia. J Neurochem 100:1121–1128
Barnes PJ (1998) Anti-inflammatory actions of glucocorticoids: molecular mechanisms. Clin Sci (Lond) 94:557–572
Pitzalis C, Pipitone N, Perretti M (2002) Regulation of leukocyte-endothelial interactions by glucocorticoids. Ann N Y Acad Sci 966:108–118
Tuckermann JP, Kleiman A, McPherson KG, Reichardt HM (2005) Molecular mechanisms of glucocorticoids in the control of inflammation and lymphocyte apoptosis. Crit Rev Clin Lab Sci 42:71–104
Van Molle W, Libert C (2005) How glucocorticoids control their own strength and the balance between pro- and anti-inflammatory mediators. Eur J Immunol 35:3396–3399
Wilckens T, De Rijk R (1997) Glucocorticoids and immune function: unknown dimensions and new frontiers. Immunol Today 18:418–424
Dhabhar FS, Miller AH, McEwen BS, Spencer RL (1996) Stress-induced changes in blood leukocyte distribution. Role of adrenal steroid hormones. J Immunol 157:1638–1644
Calandra T, Bernhagen J, Metz CN, Spiegel LA, Bacher M, Donnelly T, Cerami A, Bucala R (1995) MIF as a glucocorticoid-induced modulator of cytokine production. Nature 377:68–71
Meagher LC, Cousin JM, Seckl JR, Haslett C (1996) Opposing effects of glucocorticoids on the rate of apoptosis in neutrophilic and eosinophilic granulocytes. J Immunol 156:4422–4428
Sivertson KL, Seeds MC, Long DL, Peachman KK, Bass DA (2007) The differential effect of dexamethasone on granulocyte apoptosis involves stabilization of Mcl-1L in neutrophils but not in eosinophils. Cell Immunol 246:34–45
Liles WC, Dale DC, Klebanoff SJ (1995) Glucocorticoids inhibit apoptosis of human neutrophils. Blood 86:3181–3188
Nathan C (2002) Points of control in inflammation. Nature 420:846–852
Ravasi T, Wells C, Forest A, Underhill DM, Wainwright BJ, Aderem A, Grimmond S, Hume DA (2002) Generation of diversity in the innate immune system: macrophage heterogeneity arises from gene-autonomous transcriptional probability of individual inducible genes. J Immunol 168:44–50
Oh GS, Pae HO, Lee BS, Kim BN, Kim JM, Kim HR, Jeon SB, Jeon WK, Chae HJ, Chung HT (2006) Hydrogen sulfide inhibits nitric oxide production and nuclear factor-kappaB via heme oxygenase-1 expression in RAW264.7 macrophages stimulated with lipopolysaccharide. Free Radic Biol Med 41:106–119
Heasman SJ, Giles KM, Ward C, Rossi AG, Haslett C, Dransfield I (2003) Glucocorticoid-mediated regulation of granulocyte apoptosis and macrophage phagocytosis of apoptotic cells: implications for the resolution of inflammation. J Endocrinol 178:29–36
Yeager MP, Pioli PA, Wardwell K, Beach ML, Martel P, Lee HK, Rassias AJ, Guyre PM (2008) In vivo exposure to high or low cortisol has biphasic effects on inflammatory response pathways of human monocytes. Anesth Analg 107:1726–1734
Yona S, Gordon S (2007) Inflammation: glucocorticoids turn the monocyte switch. Immunol Cell Biol 85:81–82
Li L, Whiteman M, Moore PK (2009) Dexamethasone inhibits lipopolysacharide-induced hydrogen sulfide biosynthesis in intact cells and in an animal model of endotoxic shock. J Cell Mol Med 13:2684–2692
Lei H, Ju DW, Yu Y, Tao Q, Chen G, Gu S, Hamada H, Cao X (2000) Induction of potent antitumor response by vaccination with tumor lysate-pulsed macrophages engineered to secrete macrophage colony-stimulating factor and interferon-gamma. Gene Ther 7:707–713
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25:402–408
Jeroschewski P, Steuckart C, Kuhl M (1996) An amperometric micro-sensor for the determination of H2S in aquatic environments. Anal Chem 68:4351–4357
Doeller JE, Isbell TS, Benavides G, Koenitzer J, Patel H, Patel RP, Lancaster JR Jr, Darley-Usmar VM, Kraus DW (2005) Polarographic measurement of hydrogen sulfide production and consumption by mammalian tissues. Anal Biochem 341:40–51
Benavides GA, Squadrito GL, Mills RW, Patel HD, Isbell TS, Patel RP, Darley-Usmar VM, Doeller JE, Kraus DW (2007) Hydrogen sulfide mediates the vasoactivity of garlic. Proc Natl Acad Sci USA 104:17977–17982
Tapley D, Buettner G, Shick J (1999) Free radicals and chemiluminescence as products of the spontaneous oxidation of sulfide in seawater, and their biological implications. Biol Bull 196:52–56
Hsu DZ, Wang ST, Deng JF, Liu MY (2005) Epinephrine protects against severe acute gastric bleeding in rats: role of nitric oxide and glutathione. Shock 23:253–257
Ishii I, Akahoshi N, Yu XN, Kobayashi Y, Namekata K, Komaki G, Kimura H (2004) Murine cystathionine gamma-lyase: complete cDNA and genomic sequences, promoter activity, tissue distribution and developmental expression. Biochem J 381:113–123
Twentyman PR, Luscombe M (1987) A study of some variables in a tetrazolium dye (MTT) based assay for cell growth and chemosensitivity. Br J Cancer 56:279–285
Korhonen R, Lahti A, Hamalainen M, Kankaanranta H, Moilanen E (2002) Dexamethasone inhibits inducible nitric-oxide synthase expression and nitric oxide production by destabilizing mRNA in lipopolysaccharide-treated macrophages. Mol Pharmacol 62:698–704
Lowicka E, Beltowski J (2007) Hydrogen sulfide (H2S)—the third gas of interest for pharmacologists. Pharmacol Rep 59:4–24
Kimura H (2002) Hydrogen sulfide as a neuromodulator. Mol Neurobiol 26:13–19
Bhatia M (2005) Hydrogen sulphide is a mediator of carrageenan-induced hindpaw oedema in the rat. Br J Pharmacol 145:141–144
Zhang H, Zhi L, Moochhala SM, Moore PK, Bhatia M (2007) Endogenous hydrogen sulfide regulates leukocyte trafficking in cecal ligation and puncture-induced sepsis. J Leukoc Biol 82:894–905
Li L, Salto-Tellez M, Tan CH, Whiteman M, Moore PK (2009) GYY4137, a novel hydrogen sulfide-releasing molecule, protects against endotoxic shock in the rat. Free Radic Biol Med 47:103–113
Moilanen E, Whittle BJR, Moncada S (1999) Nitric oxide as a factor in inflammation. In: Gallin JI, Snyderman R (ed) Inflammation: principles and clinical correlations. Lippincott Williams & Wilkins, Philadelphia, pp 787–800
Zhong GZ, Chen FR, Cheng YQ, Tang CS, Du JD (2003) The role of hydrogen sulfide generation in the pathogenesis of hypertension in rats induced by inhibition of nitric oxide synthase. J Hypertens 21:1879–1885
Anuar F, Whiteman M, Siau JL, Kwong SE, Bhatia M, Moore PK (2006) Nitric oxide-releasing flurbiprofen reduces formation of proinflammatory hydrogen sulfide in lipopolysaccharide-treated rat. Br J Pharmacol 147:966–974
Walker G, Pfeilschifter J, Kunz D (1997) Mechanisms of suppression of inducible nitric-oxide synthase (iNOS) expression in interferon (IFN)-gamma-stimulated RAW 264.7 cells by dexamethasone. Evidence for glucocorticoid-induced degradation of iNOS protein by calpain as a key step in post-transcriptional regulation. J Biol Chem 272:16679–16687
Wallace JL (2007) Hydrogen sulfide-releasing anti-inflammatory drugs. Trends Pharmacol Sci 28:501–505
Cadepond F, Ulmann A, Baulieu EE (1997) RU486 (mifepristone): mechanisms of action and clinical uses. Annu Rev Med 48:129–156
Hayashi R, Wada H, Ito K, Adcock IM (2004) Effects of glucocorticoids on gene transcription. Eur J Pharmacol 500:51–62
Acknowledgments
The authors wish to thank Dr. I.Ishii, Gunma University Graduate School of Medicine, Gunma, Japan, for his gift of pCSE-PGL3 plasmid. This work was supported by the National Natural Science Foundation of China, grant nos. 30670815 and 30770846, and the Science and Technology Commission of Shanghai Municipals (09XD1405600).
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X.-Y. Zhu and S.-J. Liu contributed equally to this manuscript.
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Zhu, XY., Liu, SJ., Liu, YJ. et al. Glucocorticoids suppress cystathionine gamma-lyase expression and H2S production in lipopolysaccharide-treated macrophages. Cell. Mol. Life Sci. 67, 1119–1132 (2010). https://doi.org/10.1007/s00018-009-0250-9
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DOI: https://doi.org/10.1007/s00018-009-0250-9