Gender-based reciprocal expression of transforming growth factor-beta1 and the inducible nitric oxide synthase in a rat model of cyclophosphamide-induced cystitis.
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.
Tyagi
Pradeep
P
Department of Surgery, University of Pittsburgh, PA 15213, USA. vodovotz@upmc.edu.
Tyagi
Vikas
V
Yoshimura
Naoki
N
Witteemer
Erich
E
Barclay
Derek
D
Loughran
Patricia A
PA
Zamora
Ruben
R
Vodovotz
Yoram
Y
Brain Behav Immun. 2002 Apr;16(2):140-9
11908923
Gend Med. 2008;5 Suppl A:S3-S10
18395681
J Urol. 1999 Dec;162(6):2211-6
10569621
Am J Pathol. 2002 Jun;160(6):2081-93
12057913
Immunol Lett. 2005 Apr 15;98(1):45-8
15790507
Auton Neurosci. 2000 Oct 2;83(3):140-7
11593765
Pain. 1997 Dec;73(3):423-429
9469534
J Urol. 2008 Aug;180(2):737-41
18554637
Biol Reprod. 1998 Jun;58(6):1343-50
9623591
Am J Surg Pathol. 1990 Oct;14(10):969-76
2403198
J Cell Sci. 2003 Jan 15;116(Pt 2):217-24
12482908
Exp Toxicol Pathol. 2008 Apr;59(6):425-30
18234483
J Cell Biochem. 1993 Dec;53(4):314-22
8300749
Am J Pathol. 1999 Jun;154(6):1867-76
10362813
MedGenMed. 2007 Nov 13;9(4):34
18311384
Prog Urol. 2001 Apr;11(2):239-44
11400485
Biotechniques. 1996 Mar;20(3):390-2, 394
8679195
Urology. 2005 Nov;66(5):935-40
16286098
Urology. 2009 Feb;73(2):421-6
18848347
Miner Electrolyte Metab. 1998;24(2-3):136-43
9525696
Eur J Immunol. 2008 Jun;38(6):1689-99
18421791
Br J Urol. 1991 Jan;67(1):44-7
1704276
Am J Physiol. 1997 Feb;272(2 Pt 1):E181-90
9124321
J Gastroenterol Hepatol. 2008 Jul;23(7 Pt 1):1024-35
18505415
Biochem Pharmacol. 1979 Jul 1;28(13):2045-9
475846
Am J Physiol. 1995 May;268(5 Pt 1):L862-7
7539224
Eur J Dermatol. 2001 Jan-Feb;11(1):45-7
11174138
Ann Otol Rhinol Laryngol. 2008 Feb;117(2):145-52
18357839
Endocrine. 2005 Feb;26(1):55-63
15805586
Dig Dis Sci. 2008 Aug;53(8):2045-52
18095163
J Urol. 2007 Apr;177(4):1531-6
17382773
Transplant Proc. 2006 Jun;38(5):1314-9
16797290
Exp Biol Med (Maywood). 2008 May;233(5):620-6
18375833
Semin Nephrol. 2007 May;27(3):309-20
17533008
Kidney Int. 2004 Dec;66(6):2148-54
15569304
Cancer Res. 1999 May 1;59(9):2142-9
10232601
Urology. 2005 Dec;66(6):1349-53
16360482
J Urol. 2005 Oct;174(4 Pt 2):1704-7; discussion 1707
16148686
J Immunol. 1997 Sep 1;159(5):2418-23
9278333
Cancer Gene Ther. 2006 Jul;13(7):676-85
16470211
Am J Physiol Renal Physiol. 2007 Nov;293(5):F1678-90
17686959
J Urol. 2002 May;167(5):2229-34
11956484
J Clin Invest. 1997 Jun 15;99(12):2818-25
9185502
Cell Biol Toxicol. 2007 Sep;23(5):303-12
17225077
Crit Care Med. 2005 Dec;33(12 Suppl):S478-81
16340427
J Urol. 2001 Feb;165(2):660-6
11176454
J Urol. 2002 Jan;167(1):347-51
11743354
J Immunol. 2008 Jun 1;180(11):7327-37
18490732
Prostate. 2007 Dec 1;67(16):1825-33
17941092
Am J Hematol. 2005 Oct;80(2):166-7
16184578
Free Radic Biol Med. 1997;22(6):1007-17
9034240
J Immunol. 2008 Jun 1;180(11):7112-6
18490709
J Immunol. 2008 Feb 1;180(3):1903-12
18209088
Nitric Oxide. 1997 Feb;1(1):3-17
9701040
Prostaglandins Other Lipid Mediat. 2008 Mar;85(3-4):115-24
18194875
Nature. 1988 Jul 21;334(6179):260-2
3041283
Rheumatol Int. 2008 Sep;28(11):1161-4
18398616
Clin Exp Immunol. 2005 Nov;142(2):312-7
16232218
Curr Mol Med. 2004 Nov;4(7):787-97
15579025
J Urol. 2004 Dec;172(6 Pt 1):2261-5
15538244