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Abstract
Purpose
We investigated the effect and duration of hyaluronic acid (HA) on the bladder of rats with cyclophosphamide (CYP)-induced cystitis.
Materials and Methods
CYP (250 mg/kg) was injected intraperitoneally in female Sprague-Dawley rats to induce cystitis. HA (0.5%) was instilled intravesically. First, the rats were separated into 2 groups for urodynamic study (CYP group, CYP+HA group), and we assessed the inter-contraction interval, maximal voiding pressure (MVP), and pressure threshold of the 2 groups. Second, the rats were separated into 3 groups to assess the duration of effect of HA (CYP group; HA-1 group: HA was instilled 1 day before injection of CYP; HA-3 group: HA was instilled 3 days before injection of CYP), and we assessed the voiding interval for 7 days after confining the rats in metabolic cages.
Results
The inter-contraction interval of the CYP+HA group (236.0±18.3 min) was significantly longer than that of the CYP group (178.0±13.4 min; p<0.03), but the MVPs of both groups did not differ significantly (32.3±1.2 vs. 34.0±2.4 cmH2O, respectively; p=0.56). The pressure threshold of the CYP+HA group (9.9±0.5 cmH2O) was significantly longer than that of the CYP group (13.2±0.7 cmH2O; p=0.002). The voiding interval decreased in each of the 3 groups according to the time after cystitis had been induced. The voiding interval was maximally decreased on the second day and returned to the pre-CYP injection level on the fifth day in each group. There was a significant difference in voiding interval among the 3 groups on the second day (p<0.05). There was no significant difference between the CYP+HA-1 and CYP+HA-3 groups on the second day (p>0.05).
Figures and Tables
Fig. 1
(A) Inter-contraction interval (ICI) after instillation of cyclophosphamide (CYP). ICI of CYP+hyaluronic acid intravesical instillation (HA) group was significantly longer than the ICI of CYP group (p=0.03)a. (B) Maximal voiding pressure (MVP) after instillation of cyclophosphamide. There was no significant difference of MVP between CYP group and HA+CYP group (p=0.56)b. (C) Pressure threshold (PT) after instillation of cyclophosphamide. There was significant difference of PT between the HA+CYP group and the CYP group (p=0.002)c. No. of each animal group (CYP, CYP+HA)=11, NS: not significant.
Fig. 2
Voiding interval (VI) changes after intraperitoneal injection of cyclophosphamide (CYP), checked in the metabolic cage. Voiding interval between pre-CYP injection and each 0-8, 9-24, 48, 72 hour group was decreased significantly (ap<0.05), however, 96, 120 hour group was not decreased significantly (bp>0.05, paired t-test) in CYP, hyaluronic acid intravesical instillation (HA)-1, and HA-3 group. On the second day after CYP injection, the VI of the CYP group was significantly shorter than HA-1 group (n=6) and HA-3 group (n=6) (cp<0.05), however, there was no difference between HA-1 and HA-3 group in VI. There was no significant difference in VI between the HA-3 group and the CYP group after the third day (p>0.05). HA-1: hyaluronic acid intravesical instillation a day before CYP intraperitoneal injection, HA-3: hyaluronic acid intravesical instillation 3 days before CYP intraperitoneal injection.
Fig. 3
(A, B) Control group: normal urothelium in rat bladder. (C, D) Cyclophosphamide (CYP) group (n=11): bladder 1 day after injection of CYP. Totally ulcerated urothelium with marked edema in the lamina propria. (E, F) CYP+hyaluronic acid intravesical instillation (HA) group (n=11): bladder 1 day after injection of cyclophosphamide. Focally denuded urothelium with focal edema and hemorrhage in the lamina propria (H&E stain, A, C, E ×40; B, D, F ×200).
Fig. 4
Photomicrographs of bladder 7 days after injection of cyclophosphamide (CYP). After injection, voiding intervals of rats were checked for 5 days in the metabolic cage. One week after injection, changes in the bladders were confirmed by biopsy. (A, B) CYP group (n=6): hyperplastic changes in the urothelium and severe hemorrhages in the lamina propria and muscle layer. (C, D) hyaluronic acid intravesical instillation (HA)-1 group (n=6): hyperplastic changes in the urothelium and focal infiltration of neutrophils (less than 10/HPF) and lymphocytes (less than 10/HPF) in the lamina propria. (E, F) HA-3 group (n=6): hyperplastic changes in the urothelium and diffuse neutrophilic (more than 50/HPF) and lymphocytic (more than 100/HPF) infiltration in the lamina propria (H&E stain, A, C, E ×40; B, D, F ×200). HPF: high power field (count under ×400).
Table 1
Changes in the voiding interval (VI) after intraperitoneal injection of cyclophosphamide in the 3 groups (CYP, HA-1, HA-3)
CYP: cyclophosphamide, HA-1: hyaluronic acid intravesical instillation a day before CYP intraperitoneal injection, HA-3: hyaluronic acid intravesicalinstillation 3 days before CYP intraperitoneal injection, PRE: VI before CYP intraperitoneal injection as a control. Each VI represents Mean±SE, a: performed by Duncan multivariated analysis (CYP vs. HA-1 and CYP vs. HA-2 have a significant difference, however, HA-1 vs. HA-3 has no significant difference)
References
1. Moldwin RM, Sant GR. Interstitial cystitis: a pathophysiology and treatment update. Clin Obstet Gynecol. 2002. 45:259–272.
2. Nickel JC, Emerson L, Cornish J. The bladder mucus (glycosaminoglycan) layer in interstitial cystitis. J Urol. 1993. 149:716–718.
3. Spanos C, Pang X, Ligris K, Letourneau R, Alferes L, Alexacos N, et al. Stress-induced bladder mast cell activation: implications for interstitial cystitis. J Urol. 1997. 157:669–672.
4. Bouchelouche K, Nordling J. Recent developments in the management of interstitial cystitis. Curr Opin Urol. 2003. 13:309–313.
5. Boucher WS, Letourneau R, Huang M, Kempuraj D, Green M, Sant GR, et al. Intravesical sodium hyaluronate inhibits the rat urinary mast cell mediator increase triggered by acute immobilization stress. J Urol. 2002. 167:380–384.
6. Oberpenning F, van Ophoven A, Hertle L. Interstitial cystitis: an update. Curr Opin Urol. 2002. 12:321–332.
7. Morales A, Emerson L, Nickel JC, Lundie M. Intravesical hyaluronic acid in the treatment of refractory interstitial cystitis. J Urol. 1996. 156:45–48.
8. Porru D, Campus G, Tudino D, Valdes E, Vespa A, Scarpa RM, et al. Results of treatment of refractory interstitial cystitis with intravesical hyaluronic acid. Urol Int. 1997. 59:26–29.
9. Constantinides C, Manousakas T, Nikolopoulos P, Stanitsas A, Haritopoulos K, Giannopoulos A. Prevention of recurrent bacterial cystitis by intravesical administration of hyaluronic acid: a pilot study. BJU Int. 2004. 93:1262–1266.
10. Kyker KD, Coffman J, Hurst RE. Exogenous glycosaminoglycans coat damaged bladder surfaces in experimentally damaged mouse bladder. BMC Urol. 2005. 5:4.
11. Takahashi K, Takeuchi J, Takahashi T, Miyauchi S, Horie K, Uchiyama Y. Effects of sodium hyaluronate on epithelial healing of the vesical mucosa and vesical fibrosis in rabbits with acetic acid induced cystitis. J Urol. 2001. 166:710–713.
12. Cox PJ. Cyclophosphamide cystitis--identification of acrolein as the causative agent. Biochem Pharmacol. 1979. 28:2045–2049.
13. Hu VY, Malley S, Dattilio A, Folsom JB, Zvara P, Vizzard MA. COX-2 and prostanoid expression in micturition pathways after cyclophosphamide-induced cystitis in the rat. Am J Physiol Regul Integr Comp Physiol. 2003. 284:R574–R585.
14. Lanteri-Minet M, Bon K, de Pommery J, Michiels JF, Menetrey D. Cyclophosphamide cystitis as a model of visceral pain in rats: model elaboration and spinal structures involved as revealed by the expression of c-Fos and Krox-24 proteins. Exp Brain Res. 1995. 105:220–232.
15. Bon K, Lichtensteiger CA, Wilson SG, Mogil JS. Characterization of cyclophosphamide cystitis, a model of visceral and referred pain, in the mouse: species and strain differences. J Urol. 2003. 170:1008–1012.
16. Boucher M, Meen M, Codron JP, Coudore F, Kemeny JL, Eschalier A. Cyclophosphamide-induced cystitis in freely-moving conscious rats: behavioral approach to a new model of visceral pain. J Urol. 2000. 164:203–208.
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