Cellular and Molecular Life Sciences

, Volume 67, Issue 7, pp 1119–1132 | Cite as

Glucocorticoids suppress cystathionine gamma-lyase expression and H2S production in lipopolysaccharide-treated macrophages

  • Xiao-Yan Zhu
  • Shu-Juan Liu
  • Yu-Jian Liu
  • Shan Wang
  • Xin Ni
Research Article


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.


Glucocorticoids Hydrogen sulfide Cystathionine γ-lyase Lipopolysaccharide Macrophages 



Hydrogen sulfide


Nitric oxide


Carbon monoxide


Cystathionine gamma-lyase


Cystathionine β-synthetase




Tumor necrosis factor-α




Glucocorticoid receptor


Migration inhibitory factor








3-[4, 5-Dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide




NG-nitro-l-arginine methyl ester


Inducible nitric oxide synthase


Charcoal-stripped FBS




Interfering RNA


CSE-overexpression RAW264.7 cell lines


CSE-knockdown RAW264.7 cell lines


Tris-buffered saline/Tween 20



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).


  1. 1.
    Moore PK, Bhatia M, Moochhala S (2003) Hydrogen sulfide: from the smell of the past to the mediator of the future? Trends Pharmacol Sci 24:609–611CrossRefPubMedGoogle Scholar
  2. 2.
    Wang R (2002) Two’s company, three’s a crowd: can H2S be the third endogenous gaseous transmitter? FASEB J 16:1792–1798CrossRefPubMedGoogle Scholar
  3. 3.
    Bhatia M (2005) Hydrogen sulfide as a vasodilator. IUBMB Life 57:603–606CrossRefPubMedGoogle Scholar
  4. 4.
    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 317:30–37CrossRefPubMedGoogle Scholar
  5. 5.
    Zhao W, Zhang J, Lu Y, Wang R (2001) The vasorelaxant effect of H2S as a novel endogenous gaseous KATP channel opener. EMBO J 20:6008–6016CrossRefPubMedGoogle Scholar
  6. 6.
    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 19:1196–1198PubMedGoogle Scholar
  7. 7.
    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 47:155–160CrossRefPubMedGoogle Scholar
  8. 8.
    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 81:1322–1332CrossRefPubMedGoogle Scholar
  9. 9.
    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 146:498–505CrossRefPubMedGoogle Scholar
  10. 10.
    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 19:623–625PubMedGoogle Scholar
  11. 11.
    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 90:765–768CrossRefPubMedGoogle Scholar
  12. 12.
    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 129:1210–1224CrossRefPubMedGoogle Scholar
  13. 13.
    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 20:2118–2120CrossRefPubMedGoogle Scholar
  14. 14.
    Mariggio MA, Pettini F, Fumarulo R (1997) Sulfide influence onpolymorphonuclear functions: a possible role for Ca2+ involvement. Immunopharmacol Immunotoxicol 19:393–404CrossRefPubMedGoogle Scholar
  15. 15.
    Mariggio MA, Minunno V, Riccardi S, Santacroce R, De Rinaldis P, Fumarulo R (1998) Sulfide enhancement of PMN apoptosis. Immunopharmacol Immunotoxicol 20:399–408CrossRefPubMedGoogle Scholar
  16. 16.
    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 100:1121–1128CrossRefPubMedGoogle Scholar
  17. 17.
    Barnes PJ (1998) Anti-inflammatory actions of glucocorticoids: molecular mechanisms. Clin Sci (Lond) 94:557–572Google Scholar
  18. 18.
    Pitzalis C, Pipitone N, Perretti M (2002) Regulation of leukocyte-endothelial interactions by glucocorticoids. Ann N Y Acad Sci 966:108–118CrossRefPubMedGoogle Scholar
  19. 19.
    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 42:71–104CrossRefPubMedGoogle Scholar
  20. 20.
    Van Molle W, Libert C (2005) How glucocorticoids control their own strength and the balance between pro- and anti-inflammatory mediators. Eur J Immunol 35:3396–3399CrossRefPubMedGoogle Scholar
  21. 21.
    Wilckens T, De Rijk R (1997) Glucocorticoids and immune function: unknown dimensions and new frontiers. Immunol Today 18:418–424CrossRefPubMedGoogle Scholar
  22. 22.
    Dhabhar FS, Miller AH, McEwen BS, Spencer RL (1996) Stress-induced changes in blood leukocyte distribution. Role of adrenal steroid hormones. J Immunol 157:1638–1644PubMedGoogle Scholar
  23. 23.
    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 377:68–71CrossRefPubMedGoogle Scholar
  24. 24.
    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 156:4422–4428PubMedGoogle Scholar
  25. 25.
    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 246:34–45CrossRefPubMedGoogle Scholar
  26. 26.
    Liles WC, Dale DC, Klebanoff SJ (1995) Glucocorticoids inhibit apoptosis of human neutrophils. Blood 86:3181–3188PubMedGoogle Scholar
  27. 27.
    Nathan C (2002) Points of control in inflammation. Nature 420:846–852CrossRefPubMedGoogle Scholar
  28. 28.
    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 168:44–50PubMedGoogle Scholar
  29. 29.
    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 41:106–119CrossRefPubMedGoogle Scholar
  30. 30.
    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 178:29–36CrossRefPubMedGoogle Scholar
  31. 31.
    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 107:1726–1734CrossRefPubMedGoogle Scholar
  32. 32.
    Yona S, Gordon S (2007) Inflammation: glucocorticoids turn the monocyte switch. Immunol Cell Biol 85:81–82CrossRefPubMedGoogle Scholar
  33. 33.
    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 13:2684–2692CrossRefPubMedGoogle Scholar
  34. 34.
    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 7:707–713CrossRefPubMedGoogle Scholar
  35. 35.
    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 25:402–408CrossRefPubMedGoogle Scholar
  36. 36.
    Jeroschewski P, Steuckart C, Kuhl M (1996) An amperometric micro-sensor for the determination of H2S in aquatic environments. Anal Chem 68:4351–4357CrossRefGoogle Scholar
  37. 37.
    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 341:40–51CrossRefPubMedGoogle Scholar
  38. 38.
    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 104:17977–17982CrossRefPubMedGoogle Scholar
  39. 39.
    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 196:52–56CrossRefGoogle Scholar
  40. 40.
    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 23:253–257PubMedGoogle Scholar
  41. 41.
    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 381:113–123CrossRefPubMedGoogle Scholar
  42. 42.
    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 56:279–285PubMedGoogle Scholar
  43. 43.
    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 62:698–704CrossRefPubMedGoogle Scholar
  44. 44.
    Lowicka E, Beltowski J (2007) Hydrogen sulfide (H2S)—the third gas of interest for pharmacologists. Pharmacol Rep 59:4–24PubMedGoogle Scholar
  45. 45.
    Kimura H (2002) Hydrogen sulfide as a neuromodulator. Mol Neurobiol 26:13–19CrossRefPubMedGoogle Scholar
  46. 46.
    Bhatia M (2005) Hydrogen sulphide is a mediator of carrageenan-induced hindpaw oedema in the rat. Br J Pharmacol 145:141–144CrossRefPubMedGoogle Scholar
  47. 47.
    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 82:894–905CrossRefPubMedGoogle Scholar
  48. 48.
    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 47:103–113CrossRefPubMedGoogle Scholar
  49. 49.
    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 787–800Google Scholar
  50. 50.
    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 21:1879–1885CrossRefPubMedGoogle Scholar
  51. 51.
    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 147:966–974CrossRefPubMedGoogle Scholar
  52. 52.
    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 272:16679–16687CrossRefPubMedGoogle Scholar
  53. 53.
    Wallace JL (2007) Hydrogen sulfide-releasing anti-inflammatory drugs. Trends Pharmacol Sci 28:501–505CrossRefPubMedGoogle Scholar
  54. 54.
    Cadepond F, Ulmann A, Baulieu EE (1997) RU486 (mifepristone): mechanisms of action and clinical uses. Annu Rev Med 48:129–156CrossRefPubMedGoogle Scholar
  55. 55.
    Hayashi R, Wada H, Ito K, Adcock IM (2004) Effects of glucocorticoids on gene transcription. Eur J Pharmacol 500:51–62CrossRefPubMedGoogle Scholar

Copyright information

© Birkhäuser Verlag, Basel/Switzerland 2010

Authors and Affiliations

  • Xiao-Yan Zhu
    • 1
    • 3
  • Shu-Juan Liu
    • 1
    • 3
  • Yu-Jian Liu
    • 2
  • Shan Wang
    • 1
    • 3
  • Xin Ni
    • 1
    • 3
  1. 1.Department of PhysiologySecond Military Medical UniversityShanghaiPeople’s Republic of China
  2. 2.Department of PathophysiologySecond Military Medical UniversityShanghaiPeople’s Republic of China
  3. 3.Key Laboratory Neurobiology, Ministry of EducationSecond Military Medical UniversityShanghaiPeople’s Republic of China

Personalised recommendations