title: Gender-based reciprocal expression of transforming growth factor-beta1 and the inducible nitric oxide synthase in a rat model of cyclophosphamide-induced cystitis.
abstract: BACKGROUND
  The pluripotent cytokine transforming growth factor-beta1 (TGF-beta1) is the central regulator of inducible Nitric Oxide Synthase (iNOS) that is responsible for nitric oxide (NO) production in inflammatory settings. Previous studies have implicated a role for NO, presumably derived from iNOS, in cyclophosphamide (CYP)-induced cystitis in the bladder. TGF-beta1 is produced in latent form and requires dissociation from the latency-associated peptide (LAP) to act as primary anti-inflammatory and pro-healing modulator following tissue injury in the upper urinary tract. Since the role of TGF-beta1 in lower urinary tract inflammation is currently unknown, and since gender-based differences exist in the setting of interstitial cystitis (IC), the present study examined the relationship between TGF-beta1 and iNOS/NO in the pathogenesis of CYP-induced cystitis in both male and female rats.
  
  
  METHODS
  Sprague-Dawley rats, 4 months of age, of either gender were given 150 mg/kg CYP intraperitoneally. Urinary and bladder tissue TGF-beta1 and NO reaction products (NO2-/NO3-) were quantified as a function of time following CYP. Expression of active and latent TGF-beta1 as well as iNOS in harvested bladder tissue was assessed by immunohistochemistry.
  
  
  RESULTS
  Female rats had significantly higher levels of NO2-/NO3- in urine even at baseline as compared to male rats (p < 0.001), whereas there was no gender based significant difference in urine levels of active or latent TGF-beta1 prior to CYP injection. Inflammatory and cytotoxic changes were induced by CYP in the bladder of both sexes that were accompanied by differences in the urine levels of NO2-/NO3- and TGF-beta1. Male rats responded to CYP with significantly lower levels of NO2-/NO3- and significantly higher levels of TGF-beta1 in urine (p < 0.05) as compared to females at all time points after CYP. The urine levels of NO2-/NO3- after CYP were inversely correlated to latent and active TGF-beta1 (Pearson coefficient of -0.72 and -0.69 in females and -0.89 and -0.76 in males, respectively; p < 0.01). Bladder tissue of male rats exhibited significantly higher levels of both latent and active TGF-beta1 (p < 0.01) compared to female rats after CYP. TGF-beta1 and iNOS protein was mostly localized in the urothelium.
  
  
  CONCLUSIONS
  The results of this study suggest that there exists an inverse relationship between the expression of TGF-beta1 and iNOS/NO2-/NO3- in CYP-inflamed bladder. The gender of the animal appears to magnify the differences in urine levels of TGF-beta1 and NO2-/NO3- in this inflammatory setting. These results support the hypothesis that TGF-beta1 can suppress iNOS expression associated with bladder inflammation and reduce systemic levels of NO2-/NO3-, and further suggest that this feature of TGF-beta1 can be harnessed for therapy and diagnosis of interstitial cystitis.
authors.affiliation: Department of Surgery, University of Pittsburgh, PA 15213, USA. vodovotz@upmc.edu.
authors.forename: Pradeep
authors.initials: P
authors.lastname: Tyagi
authors.forename: Vikas
authors.initials: V
authors.lastname: Tyagi
authors.forename: Naoki
authors.initials: N
authors.lastname: Yoshimura
authors.forename: Erich
authors.initials: E
authors.lastname: Witteemer
authors.forename: Derek
authors.initials: D
authors.lastname: Barclay
authors.forename: Patricia A
authors.initials: PA
authors.lastname: Loughran
authors.forename: Ruben
authors.initials: R
authors.lastname: Zamora
authors.forename: Yoram
authors.initials: Y
authors.lastname: Vodovotz
references.citation: Brain Behav Immun. 2002 Apr;16(2):140-9
references.pmid: 11908923
references.citation: Gend Med. 2008;5 Suppl A:S3-S10
references.pmid: 18395681
references.citation: J Urol. 1999 Dec;162(6):2211-6
references.pmid: 10569621
references.citation: Am J Pathol. 2002 Jun;160(6):2081-93
references.pmid: 12057913
references.citation: Immunol Lett. 2005 Apr 15;98(1):45-8
references.pmid: 15790507
references.citation: Auton Neurosci. 2000 Oct 2;83(3):140-7
references.pmid: 11593765
references.citation: Pain. 1997 Dec;73(3):423-429
references.pmid: 9469534
references.citation: J Urol. 2008 Aug;180(2):737-41
references.pmid: 18554637
references.citation: Biol Reprod. 1998 Jun;58(6):1343-50
references.pmid: 9623591
references.citation: Am J Surg Pathol. 1990 Oct;14(10):969-76
references.pmid: 2403198
references.citation: J Cell Sci. 2003 Jan 15;116(Pt 2):217-24
references.pmid: 12482908
references.citation: Exp Toxicol Pathol. 2008 Apr;59(6):425-30
references.pmid: 18234483
references.citation: J Cell Biochem. 1993 Dec;53(4):314-22
references.pmid: 8300749
references.citation: Am J Pathol. 1999 Jun;154(6):1867-76
references.pmid: 10362813
references.citation: MedGenMed. 2007 Nov 13;9(4):34
references.pmid: 18311384
references.citation: Prog Urol. 2001 Apr;11(2):239-44
references.pmid: 11400485
references.citation: Biotechniques. 1996 Mar;20(3):390-2, 394
references.pmid: 8679195
references.citation: Urology. 2005 Nov;66(5):935-40
references.pmid: 16286098
references.citation: Urology. 2009 Feb;73(2):421-6
references.pmid: 18848347
references.citation: Miner Electrolyte Metab. 1998;24(2-3):136-43
references.pmid: 9525696
references.citation: Eur J Immunol. 2008 Jun;38(6):1689-99
references.pmid: 18421791
references.citation: Br J Urol. 1991 Jan;67(1):44-7
references.pmid: 1704276
references.citation: Am J Physiol. 1997 Feb;272(2 Pt 1):E181-90
references.pmid: 9124321
references.citation: J Gastroenterol Hepatol. 2008 Jul;23(7 Pt 1):1024-35
references.pmid: 18505415
references.citation: Biochem Pharmacol. 1979 Jul 1;28(13):2045-9
references.pmid: 475846
references.citation: Am J Physiol. 1995 May;268(5 Pt 1):L862-7
references.pmid: 7539224
references.citation: Eur J Dermatol. 2001 Jan-Feb;11(1):45-7
references.pmid: 11174138
references.citation: Ann Otol Rhinol Laryngol. 2008 Feb;117(2):145-52
references.pmid: 18357839
references.citation: Endocrine. 2005 Feb;26(1):55-63
references.pmid: 15805586
references.citation: Dig Dis Sci. 2008 Aug;53(8):2045-52
references.pmid: 18095163
references.citation: J Urol. 2007 Apr;177(4):1531-6
references.pmid: 17382773
references.citation: Transplant Proc. 2006 Jun;38(5):1314-9
references.pmid: 16797290
references.citation: Exp Biol Med (Maywood). 2008 May;233(5):620-6
references.pmid: 18375833
references.citation: Semin Nephrol. 2007 May;27(3):309-20
references.pmid: 17533008
references.citation: Kidney Int. 2004 Dec;66(6):2148-54
references.pmid: 15569304
references.citation: Cancer Res. 1999 May 1;59(9):2142-9
references.pmid: 10232601
references.citation: Urology. 2005 Dec;66(6):1349-53
references.pmid: 16360482
references.citation: J Urol. 2005 Oct;174(4 Pt 2):1704-7; discussion 1707
references.pmid: 16148686
references.citation: J Immunol. 1997 Sep 1;159(5):2418-23
references.pmid: 9278333
references.citation: Cancer Gene Ther. 2006 Jul;13(7):676-85
references.pmid: 16470211
references.citation: Am J Physiol Renal Physiol. 2007 Nov;293(5):F1678-90
references.pmid: 17686959
references.citation: J Urol. 2002 May;167(5):2229-34
references.pmid: 11956484
references.citation: J Clin Invest. 1997 Jun 15;99(12):2818-25
references.pmid: 9185502
references.citation: Cell Biol Toxicol. 2007 Sep;23(5):303-12
references.pmid: 17225077
references.citation: Crit Care Med. 2005 Dec;33(12 Suppl):S478-81
references.pmid: 16340427
references.citation: J Urol. 2001 Feb;165(2):660-6
references.pmid: 11176454
references.citation: J Urol. 2002 Jan;167(1):347-51
references.pmid: 11743354
references.citation: J Immunol. 2008 Jun 1;180(11):7327-37
references.pmid: 18490732
references.citation: Prostate. 2007 Dec 1;67(16):1825-33
references.pmid: 17941092
references.citation: Am J Hematol. 2005 Oct;80(2):166-7
references.pmid: 16184578
references.citation: Free Radic Biol Med. 1997;22(6):1007-17
references.pmid: 9034240
references.citation: J Immunol. 2008 Jun 1;180(11):7112-6
references.pmid: 18490709
references.citation: J Immunol. 2008 Feb 1;180(3):1903-12
references.pmid: 18209088
references.citation: Nitric Oxide. 1997 Feb;1(1):3-17
references.pmid: 9701040
references.citation: Prostaglandins Other Lipid Mediat. 2008 Mar;85(3-4):115-24
references.pmid: 18194875
references.citation: Nature. 1988 Jul 21;334(6179):260-2
references.pmid: 3041283
references.citation: Rheumatol Int. 2008 Sep;28(11):1161-4
references.pmid: 18398616
references.citation: Clin Exp Immunol. 2005 Nov;142(2):312-7
references.pmid: 16232218
references.citation: Curr Mol Med. 2004 Nov;4(7):787-97
references.pmid: 15579025
references.citation: J Urol. 2004 Dec;172(6 Pt 1):2261-5
references.pmid: 15538244