Effects of rotenone on inducible nitric oxide synthase and cyclooxygenase-2 mRNA levels detected by real-time PCR in a rat bladder ischemia/reperfusion model

Nergiz,İdris; Başeskioğlu,Barbaros; Yenilmez,Aydın; Erkasap,Nilüfer; Can,Cavit; Tosun,Murat

doi:10.3892/etm.2012.596

Effects of rotenone on inducible nitric oxide synthase and cyclooxygenase-2 mRNA levels detected by real-time PCR in a rat bladder ischemia/reperfusion model

Authors:
- İdris Nergiz
- Barbaros Başeskioğlu
- Aydın Yenilmez
- Nilüfer Erkasap
- Cavit Can
- Murat Tosun
View Affiliations

Affiliations: Department of Urology, Eskisehir Osmangazi University Medical Faculty, Eskisehir, Turkey, Department of Physiology, Eskisehir Osmangazi University Medical Faculty, Eskisehir, Turkey, Department of Histology and Embryology, Afyon Kocatepe University Medical Faculty, Afyon, Turkey
Published online on: May 31, 2012 https://doi.org/10.3892/etm.2012.596
Pages: 344-348

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

We aimed to determine whether rotenone treatment prevents induced ischemia/reperfusion (I/R) damage in rat bladders by detecting inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) levels by real-time PCR (RT‑PCR). A total of 18 Sprague-Dawley albino rats were used in this experiment. The experimental groups each consisted of 6 rats and were treated as follows: group I, control; group II, I/R; group III, rotenone + I/R. In the control group, the rat bladders were removed by lower abdominal incision without any procedure. In the I/R group, 1 h prior to the ischemia 1 cc physiological serum was administered and the abdominal aortas were clamped for 1 h to achieve bladder ischemia. Following the ischemia, reperfusion was induced for 1 h and the bladders were removed. In the rotenone + I/R group, the rats were treated with 25 mg/kg rotenone intraperitoneally. The iNOS and COX-2 mRNA levels in each group were detected using RT-PCR. In the I/R group, the COX-2 levels in the bladder tissue were higher compared with the control group (P<0.05). The COX-2 levels in the rotenone-treated group were statistically lower compared with the I/R group (P<0.01). Vascularization and edema were markedly increased in the I/R group. Following rotenone treatment these were abrogated inversely to inflammation. Although iNOS levels were slightly higher in the I/R group compared with the control group, iNOS levels did not decrease and no significant difference was observed between the groups with regard to rotenone treatment (P>0.05). We suggest that rotenone may be used clinically to treat I/R damage due to its diminishing effect on COX-2 levels.

View Figures

View References

1.	Viñas JL, Sola A, Genescà M, Alfaro V, Pí F and Hotter G: NO and NOS isoforms in the development of apoptosis in renal ischemia/reperfusion. Free Radic Biol Med. 40:992–1003. 2006.PubMed/NCBI
2.	Levin RM, Leggett R, Whitbeck C and Horan P: Effect of calcium and calcium chelator on the response of the bladder to in vitro ischaemia. Br J Urol. 82:882–887. 1998. View Article : Google Scholar : PubMed/NCBI
3.	Ohnishi N, Liu SP, Horan P and Levin RM: Effect of repetitive stimulation on the contractile response of rabbit urinary bladder subjected to in vitro hypoxia or in vitro ischemia followed by reoxygenation. Pharmacology. 57:139–147. 1998. View Article : Google Scholar : PubMed/NCBI
4.	Wang JH, Chen HS, Wang T, Tian KL and Diao YF: Role of oxygen-derived free radicals in superior mesenteric artery occlusion shock in rats. Chin Med J (Engl). 103:278–282. 1990.PubMed/NCBI
5.	Lin AT, Chen KK, Yang CH and Chang LS: Mannitol facilitates rabbit urinary bladder recovery from overdistension injury. Urology. 56:702–707. 2000. View Article : Google Scholar : PubMed/NCBI
6.	Saito M and Miyagawa I: Bladder dysfunction after acute urinary retention in rats. J Urol. 165:1745–1747. 2001. View Article : Google Scholar : PubMed/NCBI
7.	Greenland JE and Brading AF: Urinary bladder blood flow changes during the micturition cycle in a conscious pig model. J Urol. 156:1858–1861. 1996. View Article : Google Scholar : PubMed/NCBI
8.	Saito M and Miyagawa I: N(G)-nitro-L-arginine methylester, a nitric oxide synthase inhibitor, diminishes apoptosis induced by ischemia-reperfusion in the rat bladder. Neurourol Urodyn. 21:566–571. 2002. View Article : Google Scholar
9.	Dupouy VM, Ferre PJ, Uro-Coste E and Lefebvre HP: Time course of COX-1 and COX-2 expression during ischemia-reperfusion in rat skeletal muscle. J Appl Physiol. 100:233–239. 2006. View Article : Google Scholar : PubMed/NCBI
10.	Conners W, Whitebeck C, Chicester P, Legget L, Lin AD, Johnson A, Kogan B, Levin R and Mannikarottu A: L-NAME, a nitric oxide synthase inhibitor, diminishes oxidative damage in urinary bladder partial outlet obstruction. Am J Physiol Renal Physiol. 290:357–363. 2006. View Article : Google Scholar : PubMed/NCBI
11.	Feldkamp T, Kribben A, Roeser NF, Senter RA, Kemner S, Venkatachalam MA, Nissim I and Weinberg JM: Preservation of complex I function during hypoxia-reoxygenation-induced mitochondrial injury in proximal tubules. Am J Physiol Renal Physiol. 286:F749–F759. 2004. View Article : Google Scholar : PubMed/NCBI
12.	Sobajima S, Shimer AL, Chadderdon RC, Kompel JF, Kim JS, Gilbertson LG and Kang JD: Quantitative analysis of gene expression in a rabbit model of intervertebral disc degeneration by real-time polymerase chain reaction. Spine J. 5:14–23. 2005. View Article : Google Scholar : PubMed/NCBI
13.	Gill HS, Monson FC, Wein AJ, Ruggieri MR and Levin RM: The effects of short term in-vivo ischemia on the contractile function of the rabbit urinary bladder. J Urol. 139:1350–1354. 1988.PubMed/NCBI
14.	Bratslavsky G, Kogan BA, Matsumoto S, Aslan AR and Levin RM: Reperfusion injury of the rat bladder is worse than ischemia. J Urol. 170:2086–2090. 2003. View Article : Google Scholar : PubMed/NCBI
15.	Ichikawa H, Takagi T, Uchiyama K, Higashihara H, Katada K, Isozaki Y, Naito Y, Yoshida N and Yoshikawa T: Rotenone, a mitochondrial electron transport inhibitor, ameliorates ischemiareperfusion-induced intestinal mucosal damage in rats. Redox Rep. 9:313–316. 2004. View Article : Google Scholar
16.	Vanden Hoek TL, Shao Z, Li C, Schumacker PT and Becker LB: Mitochondrial electron transport can become a significant source of oxidative injury in cardiomyocytes. J Mol Cell Cardiol. 29:2441–2450. 1997.PubMed/NCBI
17.	Hiratsuka T, Futagami S, Tatsuguchi A, et al: COX-1 and COX-2 conversely promote and suppress ischemia-reperfusion gastric injury in mice. Scand J Gastroenterol. 40:903–913. 2005. View Article : Google Scholar : PubMed/NCBI
18.	Matsuyama M, Nakatani T, Hase T, et al: The expression of cyclooxygenases and lipoxygenases in renal ischemia-reperfusion injury. Transplant Proc. 36:1939–1942. 2004. View Article : Google Scholar : PubMed/NCBI
19.	Tetsuka T, Baier LD and Morrison AR: Antioxidants inhibit interleukin-1-induced cyclooxygenase and nitric-oxide synthase expression in rat mesangial cells. Evidence for post-transcriptional regulation. J Biol Chem. 271:11689–11693. 1996. View Article : Google Scholar