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    Korean J Urol. 2006 Feb;47(2):189-194. Korean.
    Published online Feb 28, 2006.
    https://doi.org/10.4111/kju.2006.47.2.189
    Copyright © 2006 The Korean Urological Association
    Original Article

    The Protein Expressions of Apoptosis-associated Genes in the Obstructed Ureters of Rats

    Hyeong Gon Kim, Sung Hyun Paick, Yong Soo Lho, Hyun Hoe Kim,1 and Chul Kwak1
      • Department of Urology, Konkuk University College of Medicine, Seoul, Korea.
      • 1Department of Urology, Seoul National University College of Medicine, Seoul, Korea.
    • Corresponding author (Email: hhkim@snu.ac.kr )
    Received June 13, 2005; Accepted September 30, 2005.

    Abstract

    Purpose

    The role of apoptosis in the pathogenesis of ureteral damage from obstructive uropathy has rarely been studied. This study was performed to determine the protein expression of the apoptosis-associated genes in the pathogenesis of ureteral damage during the course of obstructive uropathy in ligated rat ureters.

    Materials and Methods

    After unilateral ligation of the ureter, each group of five Sprague-Dawley rats was sacrificed and examined at 1, 5, 10, 15, 20, 25, 30 and 35 days after ligation: five rats with normal ureters were also examined as controls. The protein expressions of the fas-associated death domain (FADD), Bax, Bcl-xL and cyclooxygenases (COX)-2 genes in obstructed ureters were assessed by performing Western blotting.

    Results

    The expressions of FADD protein in the 20 and 25 day-obstructed ureters (DOUs) were significantly higher than that in control ureters and the peak was reached in the 25 DOUs. The expressions of Bcl-xL protein in the 20, 25 and 30 DOUs were significantly higher than that in the control ureters and the peak was reached in the 25 DOUs. The expression of COX-2 protein in the 5, 10, 15, 25 DOUs were significantly higher than that in the control ureters and the peak was reached in the 10 DOUs.

    Conclusions

    The FADD and Bcl-xL genes were involved in apoptosis of the obstructed ureter. The peaks of their expressions were at 25 DOUs. The expression of the COX-2 gene may be related with apoptosis in the obstructed ureter.

    Keywords
    Ureteral obstruction; Apoptosis; Rats

    Figures

    Fig. 1
    Fas-associated death domain (FADD) protein expression in the completely obstructed rat uretes. (A) Protein is extracted from the ureters before and at several time points after obstruction, and it is analyzed for the FADD and actin levels by Western blotting. (B) The relative amount of FADD protein is normalized against actin protein. The expressions of FADD protein in the 20, 25 day-obstructed ureters (DOUs) are significantly higher than that in the control ureters and the peak was reached in the 25 DOUs. *: p<0.05.

    Fig. 2
    The Bcl-xL protein expression in the completely obstructed rat ureters. (A) Protein is extracted from ureters before and at several time points after obstruction, and it is analyzed for the Bcl-xL and actin levels by Western blotting. (B) The relative amount of Bcl-xL protein is normalized against actin protein. The expressions of Bcl-xL protein in the 20, 25 and 30 DOUs are significantly higher than that in the control ureters and the peak is reached in the 25 DOUs. *: p<0.05.

    Fig. 3
    Cyclooxygenases (COX)-2 protein expression in the completely obstructed rat ureters. (A) Protein is extracted from ureters before and at several time points after obstruction, and it is analyzed for the COX-2 and actin levels by Western blotting. (B) The relative amount of COX-2 protein is normalized against actin protein. The expressions of COX-2 protein in the 5, 10, 15 and 25 DOUs is significantly higher than that in the control ureters and the peak is reached in the 10 DOUs. *: p<0.05, †: p>0.05.

    References

      1. Gobe GC, Axelsen RA. Genesis of renal tubular atrophy in experimental hydronephrosis in the rat. Role of apoptosis. Lab Invest 1987;56:273–281.
      1. Nagle RB, Bulger RE. Unilateral obstructive nephropathy in the rabbit. II. Late morphologic changes. Lab Invest 1978;38:270–278.
      1. Truong LD, Sheikh-Hamad D, Chakraborty S, Suki WN. Cell apoptosis and proliferation in obstructive uropathy. Semin Nephrol 1998;18:641–651.
      1. Choi YJ, Baranowska-Daca E, Nguyen V, Koji T, Ballantyne CM, Sheikh-Hamad D, et al. Mechanism of chronic obstructive uropathy: increased expression of apoptosis-promoting molecules. Kidney Int 2000;58:1481–1491.
      1. Truong LD, Petrusevska G, Yang G, Gurpinar T, Shappell S, Lechago J, et al. Cell apoptosis and proliferation in experimental chronic obstructive uropathy. Kidney Int 1996;50:200–207.
      1. Truong LD, Choi YJ, Tsao CC, Ayala G, Sheikh-Hamad D, Nassar G, et al. Renal cell apoptosis in chronic obstructive uropathy: the roles of caspases. Kidney Int 2001;60:924–934.
      1. Kim HG, Kwak C, Kim HH, Paick SH, Lho YS, Lee JW, et al. The serial microscopic changes of cell proliferative and apoptotic phenomenon in obstructed ureters in the rat. Korean J Urol 2005;46:495–501.
      1. Chuang YH, Chuang WL, Huang CH. Myocyte apoptosis in the pathogenesis of ureteral damage in rats with obstructive uropathy. Urology 2001;58:463–470.
      1. Chuang YH, Chuang WL, Huang SP, Huang CH. Over-expression of apoptosis-related proteins contributes to muscular damage in the obstructed ureter of the rat. BJU Int 2002;89:106–112.
      1. Chuang YH, Chuang WL, Huang SP, Huang CH. Release of cytochrome c and activation of caspases related to myocyte apoptosis in obstructed ureters in a rat model of obstructive uropathy. BJU Int 2003;92:113–118.
      1. Steller H. Mechanisms and genes of cellular suicide. Science 1995;267:1445–1449.
      1. Hu Y, Benedict MA, Ding L, Nunez G. Role of cytochrome c and dATP/ATP hydrolysis in Apaf-1-mediated caspase-9 activation and apoptosis. EMBO J 1999;18:3586–3595.
      1. Shimizu S, Narita M, Tsujimoto Y. Bcl-2 family proteins regulate the release of apoptogenic cytochrome c by the mitochondrial channel VDAC. Nature 1999;399:483–487.
      1. Li H, Yuan J. Deciphering the pathways of life and death. Curr Opin Cell Biol 1999;11:261–266.
      1. Cao Y, Prescott SM. Many actions of cyclooxygenase-2 in cellular dynamics and in cancer. J Cell Physiol 2002;190:279–286.
      1. Trifan OC, Hla T. Cyclooxygenase-2 modulates cellular growth and promotes tumorigenesis. J Cell Mol Med 2003;7:207–222.
      1. Kismet K, Akay MT, Abbasoglu O, Ercan A. Celecoxib: a potent cyclooxygenase-2 inhibitor in cancer prevention. Cancer Detect Prev 2004;28:127–142.
      1. Ding H, Han C, Zhu J, Chen CS, D'Ambrosio SM. Celecoxib derivertives induce apoptosis via the disruption of mitochondrial membrane potential and activation of caspase 9. Int J Cancer 2005;113:803–810.
      1. Chuang YH, Chuang WL, Huang SP, Liu KM, Huang CH. The temporal relationship between the severity of hydroureter and the dynamic changes of obstructed ureters in a rat model. Br J Urol 1995;76:303–310.
      1. Chinnaiyan AM, Tepper CG, Seldin MF, O'Rourke K, Kischkel FC, Hellbardt S, et al. FADD/MORT1 is a common mediator of CD95 (Fas/APO-1) and tumor necrosis factor receptor-induced apoptosis. J Biol Chem 1996;271:4961–4965.
      1. Kim PK, Dutra AS, Chandrasekharappa SC, Puck JM. Genomic structure and mapping of human FADD, an intracellular mediator of lymphocyte apoptosis. J Immunol 1996;157:5461–5466.
      1. Strasser A, Newton K. FADD/MORT1, a signal transducer that can promote cell death or cell growth. Int J Biochem Cell Biol 1999;31:533–537.
      1. Kluck RM, Bossy-Wetzel E, Green DR, Newmeyer DD. The release of cytochrome c from mitochondria: a primary site for Bcl-2 regulation of apoptosis. Science 1997;275:1132–1136.
      1. Oltvai ZN, Korsmeyer SJ. Checkpoints of dueling dimers foil death wishes. Cell 1994;79:189–192.
      1. Kujubu DA, Fletcher BS, Varnum BC, Lim RW, Herschman HR. TIS10, a phorbol ester tumor promoter-inducible mRNA from Swiss 3T3 cell, encodes a novel prostaglandin synthase/cyclooxygenase homologue. J Biol Chem 1991;266:12866–12872.
      1. Pruthi RS, Derksen E, Gaston K. Cyclooxygenase-2 as a potential target in the prevention and treatment of genitourinary tumors: a review. J Urol 2003;169:2352–2359.
      1. Tsujii M, Kawano S, Tsuji S, Sawaoka H, Hori M, DuBois RN. Cyclooxygenase regulates angiogenesis induced by colon cancer cells. Cell 1998;93:705–716.
      1. Fosslien E. Review: molecular pathology of cyclooxygenase-2 in cancer-induced angiogenesis. Ann Clin Lab Sci 2001;31:325–348.
      1. Sheng H, Shao J, Morrow JD, Beauchamp RD, DuBois RN. Modulation of apoptosis and Bcl-2 expression by prostaglandin E2 in human colon cancer cells. Cancer Res 1998;58:362–366.
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    MeSH Terms
    Animals
    Apoptosis
    Blotting, Western
    Ligation
    Prostaglandin-Endoperoxide Synthases
    Rats*
    Rats, Sprague-Dawley
    Ureter*
    Ureteral Obstruction
    bcl-X Protein
    Metrics
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