Effect of a Prolyl 4-hydroxylase Inhibitor on Bladder Fibrosis in a Rat Model of Partial Bladder Outlet Obstruction

Jae Min Chung; Dong Gil Shin; Min Jung Jung; Sang Don Lee; Jeong Zoo Lee

doi:10.4111/kju.2008.49.3.227

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Chung, Shin, Jung, Lee, and Lee: Effect of a Prolyl 4-hydroxylase Inhibitor on Bladder Fibrosis in a Rat Model of Partial Bladder Outlet Obstruction

Original Article

Korean J Urol 2008;49(3):227-236.

Published online: 17 January 2008

DOI: https://doi.org/10.4111/kju.2008.49.3.227

Effect of a Prolyl 4-hydroxylase Inhibitor on Bladder Fibrosis in a Rat Model of Partial Bladder Outlet Obstruction

Jae Min Chung¹, Dong Gil Shin², Min Jung Jung³, Sang Don Lee^4,^5,, Jeong Zoo Lee^4,⁵

^¹Departments of Urology, College of Medicine, Busan, Korea

^²Department of Urology, Moonhwa Hospitals, College of Medicine, Busan, Korea

^³Pathology, College of Medicine, Kosin, College of Medicine, Busan, Korea

^⁴Department of Urology, College of Medicine, Busan, Korea

^⁵Medical Research Institute, Pusan National University, College of Medicine, Busan, Korea

교신저자: 이상돈 부산대학교 의과대학 비뇨기과학교실 부산시 서구 아미동 1가 10 602-739, TEL: 051-240-7348, FAX: 051-247-5443, E-mail: lsd@pusan.ac.kr

Revised 14 January 2008 Accepted 4 February 2008

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Purpose

This study was performed to investigate prolyl 4-hydroxylase (P4H) expression changes and an inhibitor-mediated effect on bladder fibrosis in rats with partial bladder outlet obstruction (PBOO).

Materials and Methods

Twenty female Sprague-Dawley rats were divided into four groups. Group A (n=5) consisted of rats with PBOO treated with 2mg/kg P4H inhibitor, group B (n=5) consisted of rats with PBOO treated with 20mg/kg P4H inhibitor, group C (n=5) consisted of rats with PBOO treated with normal saline and group D (n=5) consisted of normal control animals. After PBOO for two weeks in the A, B, and C group rats, each amount of inhibitor was administered orally once a day for two weeks. After a total of four weeks, the bladders from all of the group rats were removed and evaluated.

Results

The muscle thickness calculated from Masson's trichrome staining was 0.85±0.22mm, 1.06±0.15mm, 1.19±0.30 and 0.49±0.10mm for group A, B, C, and D rats, respectively. The overall P4H expression was 65.7±15.2%, 13.4±8.4%, 73.8±15.5% and 10.0±10.0% for group A, B, C, and D rats, respectively. The overall collagen I protein expression was 16.9±18.0%, 17.0±24.1%, 30.5±13.4% and 8.8±8.7% for group A, B, C, and D rats, respectively. The overall collagen III protein expression was 9.6±4.2%, 8.8±2.9%, 12.5±10.6% and 7.5±3.5% for group A, B, C, and D rats, respectively. These results showed that PBOO led to increased muscle thickness and to an increased expression of P4H, collagen I and III protein, as compared with the group D control animals. Muscle thickness and expression of P4H, collagen I and III protein was decreased in rats treated with the P4H inhibitor-treated groups decreased as compared with rats in group C (saline-treated animals). The ratio of collagen I/III was 1.8, 1.9, 2.4 and 1.2 in group A, B, C, and D rats, respectively.

Conclusions

Our results suggest that the P4H inhibitor may be potentially utilized to reduce bladder fibrosis caused by PBOO.

Keywords: Bladder, Obstruction, Rats, Prolyl 4-hydroxylase inhibitor

REFERENCES

1. Ishimaru T, Kanamaru T, Takahashi T, Okazaki H. Inhibition of prolyl hydroxylase activity and collagen biosynthesis by fibrostatin C, a novel inhibitor produced by streptomyces catenulae subsp. griseospora No. 23924. J Antibiot. 1988; 41:1668–74.

2. Bickel M, Baader E, Brocks DG, Engelbart K, Gunzler V, Schmidts HL, et al. Beneficial effects of inhibitors of prolyl 4-hydroxylase in CCl4-induced fibrosis of the liver in rats. J Hepatol. 1991; 13(Suppl 3):S26–33.

3. Hurych J, Chvapil M, Tichy M, Beniac F. Evidence for a faster degradation of an atypical hydroxyproline and hydroxylysine deficient collagen formed under the effect of 2,2'-dipyridyl. Eur J Biochem. 1967; 3:242–7.

4. Kasper M, Fuller SD, Schuh D, Müller M. Immunohistological detection of the beta subunit of prolyl 4-hydroxylase in rat and mini pig lungs with radiation-induced pulmonary fibrosis. Virchows Arch. 1994; 425:513–9.

5. Matsui H, Kubochi K, Okazaki I, Yoshino K, Ishibiki K, Kitajima M. Collagen biosynthesis in gastric cancer: immunohistochemical analysis of prolyl 4-hydroxylase. J Surg Oncol. 1999; 70:239–46.

6. Sasaki T, Holeyfield KC, Uitto J. Doxorubicin-induced inhibition of prolyl hydroxylation during collagen biosynthesis in human skin fibroblast cultures. Relevance to imparied wound healing. J Chin Invest. 1987; 80:1735–41.

7. Majamaa K, Sasaki T, Uitto J. Inhibition of prolyl hydroxylation during collagen biosynthesis in human skin fibroblast cultures bt ethyl 3,4-dihydroxybenzoate. J Invest Dermatol. 1987; 89:405–9.

8. Karvonen K, Ala-Kokko L, Pihlajaniemi T, Helaakoski T, Henke S, Günzler V, et al. Specific inactivation of prolyl 4-hydroxylase and inhibition of collagen synthesis by oxapro-line-containing peptides in cultured human skin fibroblasts. J Biol Chem. 1990; 265:8415–9.

9. Tschank G, Brocks DG, Engelbart K, Mohr J, Baader E, Günzler V, et al. Inhibition of prolyl hydroxylation and procollagen processing in chick-embryo calvaria by a derivative of pyridine-2,4-dicarboxylate. Characterization of the diethyl ester as a proinhibitor. Biochem J. 1991; 275:469–76.

10. Fuller GC. Perspectives for the use of collagen synthesis inhibitors as antifibrotic agents. J Med Chem. 1981; 24:651–8.

11. Sakaida I, Okita K. New prolyl 4-hydroxylase inhibitor reduces procollagen gene expression and enzyme-altered lesions in rat liver cirrhosis. J Gastroenterol Hepatol. 1995; 10(Suppl 1):): S63-4.

12. Böker K, Schwarting G, Kaule G, Günzler V, Schmidt E. Fibrosis of the liver in rats induced by bile duct ligation. Effects of inhibition by prolyl 4-hydroxylase. J Hepatol. 1991; 13(Suppl 3):S35–40.

13. Kawaguchi Y, Harigai M, Kitani A, Suzuki K, Kawakami M, Ishizuka T, et al. Effect of prolyl 4-hydroxylase inhibitor on fibroblast collagen production in vitro: an approach to the treatment of systemic sclerosis. J Rheumatol. 1992; 19:1710–5.

14. Lee SD, Akbal C, Miseeri R, Jung C, Rink R, Kaefer M. Collagen prolyl 4-hydroxylase is up-regulated in an acute bladder outlet obstruction. J Pediatr Urol. 2006; 2:225–32.

15. Shapiro E, Becich MJ, Perlman E, Lepor H. Bladder wall abnormalities in myelodysplastic bladders: a computer assissted morphometric analysis. J Urol. 1991; 145:1024–9.

16. Gilpin SA, Gosling JA, Barnard RJ. Morphological and morphometric studies of the human obstructed, trabeculated urinary bladder. Br J Urol. 1985; 57:525–9.

17. Noh JY, Han SW, Kim JH, Kim CS, Hong CH. What causes bladder fibrosis?: abnormal innervation or abnormal bladder dynamics. Korean J Urol. 2003; 44:1058–63.

18. Levin RM, Hass MA, Bellamy F, Horan P, Whitbeck K, Chow PH, et al. Effect of oral Tadenan treatment on rabbit bladder structure and function after partial outlet obstruction. J Urol. 2002; 167:2253–9.

19. Lee AK, Sung YS, Kim KM, Choi H, Kim SW. Early expression of collagen type I, III and IV and Matrix- Meta-lloproteinase (MMP)-2 mRNA in the rat bladder subjected to partial outlet obstruction. Korean J Urol. 1998; 39:213–22.

20. Elbadawi A. BPH-associated voiding dysfunction: detrusor is pivotal. Contemp Urol. 1994; 6:21–38.

21. Mattiasson A, Uvelius B. Changes in contractile properties in hypertrophic rat urinary bladder. J Urol. 1982; 128:1340–2.

22. Kim KM, Kogan BA, Massad CA, Huang YC. Collagen and elastin in the obstructed fetal bladder. J Urol. 1991; 146:528–31.

23. Ewalt DH, Constantinesco S, Howard PS, Blyth B, Snyder HM, Duckett JW, et al. Increased collagen type III in the neurogenic non-compliant bladder. J Urol. 1992; 147:438A.

24. Ewalt DH, Howard PS, Blyth B, Snyder HM 3rd, Duckett JW, Levin RM, et al. Is laminar propria matrix responsibie for normal bladder compliance? J Urol. 1992; 148:544–9.

25. Winter AD, Page AP. Prolyl 4-hydroxylase is an essential procollagen-modifying enzyme required for exoskeleton formation and the maintenance of body shape in the nematode Caenorhabditis elegans. Mol Cell Biol. 2000; 20:4084–93.

26. Kukkola L, Hieta R, Kivirikko KI, Myllyharju J. Identification and characterization of a third human, rat, and mouse collagen prolyl 4-hydroxylase isoenzyme. J Biol Chem. 2003; 278:47685–93.

27. Nissi R, Autio-Harmainen H, Marttila P, Sormunen R, Kivirikko KI. Prolyl 4-hydroxylase isoenzymes I and II have different expression patterns in several human tissues. J Histochem Cytochem. 2001; 49:1143–53.

28. Berggren T, Uvelius B. Cystometrical evaluation of acute and chronic overdistension in the rat urinary bladder. Urol Res. 1998; 26:325–30.

29. Mostwin JL, Karim OM, van Koeveringe G, Books EL. The guinea pig as a model of gradual urethral obstruction. J Urol. 1991; 145:854–8.

Fig. 1.

Partial bladder outlet obstruction model in rats. (A) Through a lower transverse incision, the middle urethra was exposed. (B) One surgeon consistently ligated around the urethra with 4/0 silk in the presence of an intraluminally placed medicut catheter with a steel rod and extraluminally placed medicut catheter without a steel rod.

Fig. 2.

Muscle thickness (Masson's trichrome staining, x40). (Group A) Partial bladder outlet obstruction (PBOO) in rats treated with 2 mg/kg prolyl 4-hydroxylase (P4H) inhibitor, (Group B) PBOO in rats treated with 20mg/kg P4H inhibitor, (Group C) PBOO in rats treated with normal saline, (Group D) normal control rats.

Fig. 3.

Prolyl 4-hydroxylase (P4H) expression (immunohistochemical staining, x400). (Group A) partial bladder outlet obstruction (PBOO) in rats treated with 2mg/kg P4H inhibitor, (Group B) PBOO in rats treated with 20mg/kg P4H inhibitor, (Group C) PBOO in rats treated with normal saline, (Group D) normal control rats.

Fig. 4.

Collagen I expression (immunohistochemical staining, x400). (Group A) partial bladder outlet obstruction (PBOO) in rats treated with 2mg/kg P4H inhibitor, (Group B) PBOO in rats treated with 20mg/kg P4H inhibitor, (Group C) PBOO in rats treated with normal saline, (Group D) normal control rats.

Fig. 5.

Collagen III expression (immunohistochemical staining, x400). (Group A) partial bladder outlet obstruction (PBOO) in rats treated with 2mg/kg P4H inhibitor, (Group B) PBOO in rats treated with 20mg/kg P4H inhibitor, (Group C) PBOO in rats treated with normal saline, (Group D) normal control rats.

Table 1.

Data summary of muscle thickness, P4H, collagen I and III

	Group A	Group B	Group C	Group D
Muscle thickness (mm)	0.85±0.22	1.06±0.15	1.19±0.30	0.49±0.10
P4H expression (%)	65.7±15.2	13.4±8.4	73.8±15.5	10.0±10.0
Collagen I expression (%)	16.9±18.0	17.0±24.1	30.5±13.4	8.8±8.7
Collagen III expression (%)	9.6±4.2	8.8±2.9	12.5±10.6	7.5±3.5
Collagen I/III ratio	1.8	1.9	2.4	1.2

P4H: prolyl 4-hydroxylase, Group A: partial bladder outlet obstruction (PBOO) with P4H inhibitor 2mg/kg, Group B: PBOO with P4H inhibitor 20mg/kg, Group C: PBOO with normal saline, Group D: normal control

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