Journal Mobile Options
Table of Contents
Vol. 80, No. 2, 2004
Issue release date: 2004

Tumor Necrosis Factor-Alpha-Induced Nitric Oxide Restrains the Apoptotic Response of Anterior Pituitary Cells

Candolfi M. · Jaita G. · Zaldivar V. · Zárate S. · Pisera D. · Seilicovich A.
To view the fulltext, log in and/or choose pay-per-view option

Individual Users: Register with Karger Login Information

Please create your User ID & Password

Contact Information

I have read the Karger Terms and Conditions and agree.

To view the fulltext, please log in

To view the pdf, please log in


We previously reported that tumor necrosis factor-α (TNF-α) inhibits cell proliferation whereas it stimulates apoptosis of anterior pituitary cells in an estrogen-dependent manner. Also, we showed that nitric oxide (NO) mediates the inhibitory effect of TNF-α on prolactin release. Here, we studied the effect of TNF-α on nitric oxide synthase (NOS) activity and expression in anterior pituitary cells from cycling and ovariectomized (OVX) rats, and the role of NO in TNF-α induced apoptosis of anterior pituitary cells. NOS activity was higher in anterior pituitary cells from rats in proestrus than in diestrus and was stimulated by 17β-estradiol (10–9 M, E2). TNF-α (50 ng/ml) stimulated NOS activity in anterior pituitary cells from rats at both stages of the estrous cycle and in cells from OVX rats cultured either with or without E2. Inducible NOS (iNOS) gene expression was higher in anterior pituitary cells from rats in proestrus than in diestrus and its expression was enhanced by TNF-α. Acute administration of E2 to OVX rats increased endothelial NOS (eNOS) expression in the anterior pituitary gland. Also, E2 increased eNOS mRNA in dispersed anterior pituitary cells from OVX rats, and this effect was blocked by TNF-α. nNOS expression in the anterior pituitary gland was higer at proestrus than at diestrus but eNOS expression was similar at both stages. TNF-α decreased eNOS mRNA in anterior pituitary cells from rats at proestrus or diestrus. In anterior pituitary cells from OVX rats, TNF-α failed to induce apoptosis but was able to induce it when cells were incubated with NAME or NMMA, NOS inhibitors that did not affect cell viability per se. In the presence of E2, NAME induced apoptosis and enhanced the proapoptotic effect of TNF-α. In conclusion, our study shows that TNF-α upregulates iNOS gene expression whereas it downregulates estrogen-induced eNOS expression in anterior pituitary cells. Endogenous NO may restrain rather than mediate the proapoptotic effect of TNF-α in anterior pituitary cells.

Copyright / Drug Dosage

Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.


  1. Leong KG, Karsan A: Signaling pathways mediated by tumor necrosis factor a. Histol Histopathol 2000;15:1303–1325.
  2. Nadeau S, Riviest S: Regulation of the gene encoding tumor necrosis factor alpha (TNF-alpha) in rat brain and pituitary in response in different models of systemic immune challenge. J Neuropathol Exp Neurol 1999;58:61–77.
  3. Turnbull AV, Rivier CL: Regulation of the hypothalamic-pituitary-adrenal axis by cytokines: actions and mechanism of action. Physiol Rev 1999;79:1–71.
  4. Kobayashi H, Fukata J, Murakami N, Usui T, Ebisui O, Muro S, Hanaoka I, Inoue K, Imura H, Nakao K: Tumor necrosis factor receptor in the pituitary cells. Brain Res 1997;758:45–50.
  5. Wolvers DAW, Marquette C, Berkenbosch F, Haour F: Tumor necrosis factor-a binding sites in rodent brain and pituitary gland. Eur Cytokine Netw 1993;4:377–381.
  6. Ray D, Melmed S: Pituitary cytokine and growth factor expression and action. Endocr Rev 1997;18:206–228.
  7. Theas MS, De Laurentiis A, Lasaga M, Pisera D, Duvilanski B, Seilicovich A: Effect of lipopolysaccharide on tumor necrosis factor and prolactin secretion from rat anterior pituitary cells. Endocrine 1998;8:241–245.
  8. Theas S, Pisera D, Duvilanski B, De Laurentiis A, Pampillo M, Lasaga M, Seilicovich A: Estrogens modulate the inhibitory effect of tumor necrosis factor-a on anterior pituitary growth and prolactin release. Endocrine 2000;12:249–255.
  9. Artz E, Páez Pereda M, Perez Castro C, Pagotto U, Renner U, Stalla GK: Pathophysiological role of the cytokine network in the anterior pituitary. Front Neuroendocr 1999;20:71–95.
  10. Candolfi M, Zaldivar V, De Laurentiis A, Jaita G, Pisera D, Seilicovich A: Tumor necrosis factor-a induces apoptosis of lactotropes from female rats. Endocrinology 2002;143:3611–3617.
  11. Song W, Lu X, Feng Q: Tumor necrosis factor-a induces apoptosis via inducible nitric oxide synthase in neonatal mouse cardiomyocytes. Cardiosvasc Res 2000;45:595–602.
  12. Aono K, Isobe KI, Kiuchi K, Fan ZH, Ito M, Takeuchi A, Miyachi M, Nakashima I, Nimura Y: In vitro and in vivo expression of inducible nitric oxide synthase during experimental endotoxemia: Involvement of other cytokines. J Cell Biochem 1997;65:349–358.
  13. De Vera ME, Taylor BS, Wang Q, Shapiro RA, Billiar TR, Geller DA: Dexamethasone suppresses iNOS gene expression by upregulating I-kappa B alpha and inhibiting NF-kappa B. Am J Physiol 1997;273:G1290–G1296.
  14. Theas S, De Laurentiis A, Candolfi M, López SL, Carrasco AE, Zaldivar V, Seilicovich A: Nitric oxide mediates the inhibitory effect of tumor necrosis factor-a on prolactin release. Neuroendocrinology 2001;74:82–86.
  15. Brüne B: Nitric oxide: NO apoptosis or turning it ON? Cell Death Differ 2003;10:864–869.
  16. Liu L, Stamler JS: NO: An inhibitor of cell death. Cell Death Differ 1999;6:937–942.
  17. Griffith OW, Stuehr DJ: Nitric oxide synthase: Properties and catalytic mechanism. Annu Rev Physiol 1995;57:707–736.
  18. Yun HY, Dawson VL, Dawson TM: Neurobiology of nitric oxide. Crit Rev Neurobiol 1996;10:291–316.
  19. Ceccatelli S, Hulting AL, Zhang X, Gustafsson L, Villar M, Hökfelt T: Nitric oxide synthase in rat anterior pituitary gland and the role of nitric oxide in regulation of luteinizing hormone secretion. Proc Natl Acad Sci USA 1993;11292–11296.
  20. Lloyd RV, Jin L, Qian X, Zhang SY, Scheithauer BW: Nitric oxide synthase in the human pituitary gland. Am J Pathol 1995;146:86–94.
  21. Qian X, Jin L, Lloyd RV: Estrogen downregulates neuronal nitric oxide synthase in rat anterior pituitary cells and GH3 tumors. Endocrine 1999;1:123–130.
  22. Kostic TS, Andric SA, Stojilkovic SS: Spontaneous and receptor-controlled soluble guanylyl cyclase activity in anterior pituitary cells. Mol Endocrinol 2001;15:1010- 1022.
  23. Wong ML, Rettori V, Al-Shekhlee A, Bongiorno PB, Canteros G, McCann SM, Gold PW, Licinio J: Inducible nitric oxide synthase gene expression in the brain during systemic inflammation. Nat Med 1996;2:581–584.
  24. Satta MA, Jacobs RA, Kaltsas GA, Grossman AB: Endotoxin induces Interleukin-1β and nitric oxide synthase mRNA in rat hypothalamus and pituitary. Neuroendocrinology 1998;67:109–116.
  25. Vankelecom H, Matthys P, Denef C: Inducible nitric oxide synthase in the anterior pituitary gland: Induction by interferon-γ in a subpopulation of folliculostellate cells and in an unidentifiable population of non-hormone-secreting cells. J Histochem Cytochem 1997;45:847–857.
  26. Lozach A, Garrel G, Lerrant Y, Berault A, Counis R: GnRH-dependent upregulation of nitric oxide synthase I level in pituitary gonadotrophs mediates cGMP elevation during rat proestrus. Mol Cell Endocrinol 1998;143:43–51.
  27. Garrel G, Lerrant Y, Siriostis C, Berault A, Magre S, Bouchaud C, Counis R: Evidence that gonadotropin-releasing hormone stimulates gene expression and levels of active nitric oxide synthase type I in pituitary gonadotrophs, a process altered by desensitization and, indirectly, by gonadal steroids. Endocrinology 1998;139:2163–2170.
  28. Hatano E, Bennett BL, Manning AM, Qian T, Lemasters JJ, Brenner DA: NFkappaB stimulates inducible nitric oxide synthase to protect mouse hepatocytes from TNF-alpha- and Fas-mediated apoptosis. Gastroenterology 2001;120:1251–1262.
  29. Cardaropoli S, Silvagno F, Morra E, Pescarmona GP, Todros T: Infectious and inflammatory stimuli decrease endothelial nitric oxide synthase activity in vitro. J Hypertens 2003;21:2103–2110.
  30. Anderson HD, Rahmutula D, Gardner DG: Tumor necrosis factor-(alpha) inhibits endothelial nitric-oxide synthase gene promoter activity in bovine aortic endothelial cells. J Biol Chem 2004;279:963–969.
  31. Neumann P, Gertzberg N, Johnson A: TNF-{alpha} induces a decrease in eNOS promoter activity. Am J Physiol 2004;286:L452–L459.
  32. Paz Y, Frolkis I, Pevni D, Shapira I, Yuhas Y, Iaina A, Wollman Y, Chernichovski T, Nesher N, Locker C, Mohr R, Uretzky G: Effect of tumor necrosis factor-alpha on endothelial and inducible nitric oxide synthase messenger ribonucleic acid expression and nitric oxide synthesis in ischemic and nonischemic isolated rat heart. J Am Coll Cardiol 2003;42:1299–1305.
  33. Lai PF, Mohamed F, Monge JC, Stewart DJ: Downregulation of eNOS mRNA expression by TNFalpha: Identification and functional characterization of RNA-protein interactions in the 3′UTR. Cardiovasc Res 2003;59:160–168.
  34. Ashkenazi A, Dixit VM: Death receptors: Signaling and modulation. Science 1998;281:1305–1308.
  35. Velardez MO, Poliandri AH, Cabilla JP, Bodo CC, Machiavelli LI, Duvilanski BH: Long term treatment of anterior pituitary cells with nitric oxide induces programmed cell death. Endocrinology 2004;145:2064–2070.
  36. Cirino G, Fiorucci S, Sessa WC: Endothelial nitric oxide synthase: The Cinderella of inflammation? Trends Pharmacol Sci 2003;24:91–95.
  37. Dimmeler S, Zeiher AM: Nitric oxide-an endothelial cell survival factor. Cell Death Differ 1999;6:964–968.
  38. Li J, Bombeck CA, Yang S, Kim YM, Billiar TR: Nitric oxide suppresses apoptosis via interrupting caspase activation and mitochondrial dysfunction in cultured hepatocytes. J Biol Chem 1999;274:17325–17333.
  39. Kim YM, Chung HT, Kim SS, Han JA, Yoo YM, Kim KM, Lee GH, Yun HY, Green A, Li J, Simmons RL, Billiar TR: Nitric oxide protects PC12 cells from serum deprivation-induced apoptosis by cGMP-dependent inhibition of caspase signaling. J Neurosci 1999;19:6740–6747.

Pay-per-View Options
Direct payment This item at the regular price: USD 38.00
Payment from account With a Karger Pay-per-View account (down payment USD 150) you profit from a special rate for this and other single items.
This item at the discounted price: USD 26.50