The changes of Inducible Nitric Oxide Synthase, Endothelial Nitric Oxide Synthase and Cyclooxygenase-2 mRNA expressions in intrauterine tissues of pregnant rats
Abstract
Prostaglandins (PGs) are produced by the intrauterine tissues of pregnancy and play important role in all of the physiological processes of parturition. PGs are synthesized from arachidonic acid by the action of cyclooxygenase (COX). Nitric oxide (NO) is a potent smooth muscle relaxant and is considered to be an important local mediator that suppresses uterine contractility. There may be an interaction between the NO and PG pathways in regulating the nitric oxide synthase (NOS) and COX expression and this interaction may play a critical role in the control of cervical ripening and parturition. The purpose of this study is to examine the expression of cyclooxygenase-2 (COX-2), inducible NOS (iNOS) and endothelial NOS (eNOS) mRNA levels during gestation, parturition, and postpartum in pregnant rat uterus. Forty-two pregnant Sprague Dawley rats were randomly divided into 7 groups. Both uterine horns were removed and the pups and placentas were discarded. Complementary DNA (cDNA) was synthesized after isolation of total RNA from the tissues. The relative expression of iNOS, eNOS and COX-2 genes were measured by quantitative real-time polymerase chain reaction (QRT-PCR). The mRNA level of iNOS was increased during gestation; highest level in parturition and then it decreased at postpartum. The eNOS mRNA level was generally low during gestation in all groups and was significantly increased at postpartum. The COX-2 mRNA levels decreased as the pregnancy progressed to term with the highest level occurring on day 21 and the lovest levels during the postpartum period. In conclusion, there may be an interaction between the NO and PG pathways in cervical ripening and parturition. These results may show that mRNA levels of eNOS, iNOS and COX-2 genes may be effective cervical ripening during pregnancy and parturition
References
- Abel MH, Kelly RW (1979): Differential production of prostaglandins within the human uterus. Prostaglandins, 18, 821-828.
- Ali M, Buhimschi I, Chwalisz K, Garfield RE (1997): Changes in expression of the nitric oxide synthase isoforms in rat uterus and cervix during pregnancy and parturition. Mol Hum Reprod, 3, 995-1003.
- Arslan A, Zingg HH (1996): Regulation of COX-2 gene expression in rat uterus in vivo and in vitro. Prostaglandins, 52, 463-481.
- Bansal RK, Goldsmith PC, He Y, Zaloudek CJ, Ecker JL, Riemer RK (1997): A decline in myometrial nitric oxide synthase expression is associated with labor and delivery. J Clin Invest, 99, 2502-2508.
- Buhimschi I, Ali M, Jain V, Chwalisz K, Garfield RE (1996): Differential regulation of nitric oxide in the rat uterus and cervix during pregnancy and labour. Hum Reprod, 11, 1755-1766.
- Chwalisz K, Garfield RE (1997): Regulation of the uterus and cervix during pregnancy and labor. Role of progesterone and nitric oxide. Ann N Y Acad Sci, 828, 238-53.
- Dong YL, Gangula PR, Fang L, Yallampalli C (1996): Differential expression of cyclooxygenase-1 and -2 proteins in rat uterus and cervix during the estrous cycle, pregnancy, labor and in myometrial cells. Prostaglandins, 52, 13-34.
- Dong YL, Gangula PRR, Yallampalli C (1996): Nitric oxide synthase isoforms in the rat uterus: differential regulation during pregnancy and labor. J Reprod Fertil, 107, 249-254.
- Fittkow CT, Shi SQ, Bytautiene E, Olson G, Saade GR, Garfield RE (2001): Changes in light-induced fluorescence of cervical collagen in guinea pigs during gestation and after sodium nitroprusside treatment. J Perinat Med, 29, 535-543.
- Ghabour MS, Eis AL, Brockman DE, Pollock JS, Myatt L (1995): Immunohistochemical characterization of placental nitric oxide synthase expression in preeclampsia. Am J Obstet Gynecol, 173, 687-694.
- Gibb W (1998): The role of prostaglandins in human parturition. Ann Med, 30, 235-241.
- Keirse MJ (1993): Prostaglandins in preinduction cervical ripening. Meta-analysis of worldwide clinical experience. J Reprod Med, 38, 89-100.
- Knowles RG, Moncada S (1994): Nitric oxide synthases in mammals. Biochem J, 298, 249-258.
- Ledingham MA, Thomson AJ, Young A, Macara LM, Greer IA, Norman JE (2000): Changes in the expression of nitric oxide synthase in the human uterine cervix during pregnancy and parturition. Mol Hum Reprod, 6, 1041- 1048.
- Marx SG, Wentz MJ, Mackay LB, Schlembach D, Maul H, Fittkow C, Given R, Vedernikov Y, Saade GR, Garfield RE (2006): Effects of progesterone on iNOS, COX-2, and collagen expression in the cervix. J Histochem Cytochem, 54, 623-639.
- Mitchell MD, Romero RJ, Edwin SS, Trautman MS (1995): Prostaglandins and parturition. Reprod Fertil Dev, 7, 623-632.
- Moncada SR, Palmer MG, Higgs EA (1995): Nitric oxide: physiology, pathophysiology and pharmacology. Pharmacol Rev, 43, 109-142.
- Natuzzi ES, Ursell PC, Harrison M, Buscher C, Riemer RK (1993): Nitric oxide synthase activity in the pregnant uterus decreases at parturition. Biochem Biophys Res Commun, 194, 1-8.
- Okawa T, Asano K, Takahashi H, Hashimoto S, Anbe H, Sato A, Vedernikov YP, Saade GR, Garfield RE (2004): Expression of inducible nitric oxide synthase messenger RNA, but not guanylate cyclase messenger RNA, depends on gestational age in rat myometrium. Gynecol Endocrinol, 19, 146-151.
- Olson DM, Mijovic JE, Sadowsky DW (1995): Control of human parturition. Semin Perinatol, 19, 52-63.
- Sladek SM, Regenstein AC, Lykins D, Roberts JM (1993): Nitric oxide synthase activity in pregnant rabbit uterus decreases on the last day of pregnancy. Am J Obstet Gynecol, 169, 1285-1291.
- Smith WL, DeWitt DL (1996): Prostaglandin endoperoxide H synthases-1 and -2. Adv Immunol, 62, 167-215.
- Smith WL, Garavito RM, DeWitt DL (1996): Prostaglandin endoperoxide H synthases (cyclooxygenases)-1 and -2. J Biol Chem, 271, 33157- 33160.
- Swaisgood CM, Zu HX, Perkins DJ, Wu S, Garver CL, Zimmerman PD, Iams JD, Kniss DA (1997): Coordinate expression of inducible nitric oxide synthase and cyclooxygenase-2 genes in uterine tissues of endotoxin- treated pregnant mice. Am J Obstet Gynecol, 177, 1253- 1262.
- Tetsuka T, Daphna-Iken D, Srivastava SK, Baier LD, DuMaine J, Morrison AR (1994): Cross-talk between cyclooxygenase and nitric oxide pathways: prostaglandin E2 negatively modulates induction of nitric oxide synthase by interleukin 1. Proc Natl Acad Sci USA, 91, 12168- 12172.
- Thomson AJ, Telfer JF, Kohnen G, Young A, Cameron IT, Greer IA, Norman JE (1997): Nitric oxide synthase activity and localization do not change in uterus and placenta during human parturition. Hum Reprod, 12, 2546-2552.
- Tschugguel W, Schneeberger C, Lass H, Stonek F, Zaghlula MB, Czerwenka K, Schatten C, Kaider A, Husslein P, Huber JC (1999): Human cervical ripening is associated with an increase in cervical inducible nitric oxide synthase expression. Biol Reprod, 60, 1367-1372.
- Wu WX, Ma XH, Nathanielsz PW (1999): Tissue- specific ontogenic expression of prostaglandin H synthase 2 in the ovine myometrium, endometrium, and placenta during late gestation and at spontaneous term labor. Am J Obstet Gynecol, 181, 1512-1519.
- Yang ZM, Das SK, Wang J, Sugimoto Y, Ichikawa A, Dey SK (1997): Potential sites of prostaglandin actions in the periimplantation mouse uterus: differential expression and regulation of prostaglandin receptor genes. Biology of Reproduction, 56, 368-379.
- Zhang H, Chen X, Teng X, Snead C, Catravas JD (1998): Molecular cloning and analysis of the rat inducible nitric oxide synthase gene promoter in aortic smooth muscle cells. Biochem Pharmacol, 55, 1873-1880.
- Zuo J, Lei ZM, Rao CV, Pietrantoni M, Cook VD (1994): Differential cyclooxygenase-1 and -2 gene expression in human myometria from preterm and term deliveries. J Clin Endocrinol Metab, 79, 894-899.
- Pfaffl MW, Horgan GW, Dempfle L (2002): Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Res, 30, 36.
Gebe ratların rahim içi dokularında Uyarılabilir Nitrik Oksit Sentaz, Endotelyal Nitrik Oksit Sentazve Siklooksijenaz-2 mRNA ekspresyon değişiklikleri
Abstract
Prostaglandinler (PG) gebelikte rahim içi dokularda üretilirler ve doğum sırasındaki tüm fizyolojik süreçlerde önemli rol oynarlar. PG’ler araşidonik asitten siklooksijenaz (COX) enziminin aktivitesi ile sentezlenirler. Nitrik oksit (NO) güçlü bir düz kas gevşetici ve uterus kasılmasını baskılayan önemli bir bölgesel ara bulucu olarak kabul edilir. Nitrik oksit sentaz (NOS) ve COX ifadelenmesinin düzenlenmesinde NO ve PG yolakları arasında bir etkileşim olabilir ve bu etkileşim servikal olgunlaşma ve doğumun kontrolünde kritik bir rol oynayabilir. Bu çalışmanın amacı, hamile rat uterusunda gebelikte, doğum sırasında ve doğum sonrasında siklooksijenaz-2 (COX-2), uyarılabilir NOS (iNOS) ve endotelyal NOS (eNOS) mRNA düzeylerini incelemektir. Kırk iki hamile Sprague Dawley rat rastgele olarak 7 gruba ayrıldı. Her iki uterin boynuzu ve yavrular ve plasenta çıkarıldı. Tamamlayıcı DNA (cDNA) dokulardan total RNA izole edildikten sonra sentezlendi. iNOS, eNOS ve COX-2 genlerinin göreceli ifadelenmesi kantitatif gerçek zamanlı polimeraz zincir reaksiyonu (QRT-PCR) ile ölçüldü. iNOS mRNA düzeyleri gebelik sırasında arttı, doğum sırasında en yüksek düzeye çıktı ve doğum sonrasında azaldı. eNOS mRNA düzeyleri tüm gruplarda gebelik boyunca genel olarak düşüktü ve doğum sonrasında belirgin bir şekilde yükseldi. COX-2 mRNA düzeyleri gebeliğin sonlarına doğru azaldı, gebeliğin 21. gününde en yüksek seviyeye ulaştı ve doğum sonrası dönemde en düşük düzeylerde kaldı. Sonuç olarak servikal genişleme ve doğum sırasında, NO ve PG yolakları arasında bir etkileşim olabilir. Bu sonuçlar eNOS, iNOS ve COX-2 genlerinin mRNA düzeylerindeki değişikliklerin gebelikte servikal olgunlaşma ve doğum sırasında etkili olabileceğini gösterebilir
References
- Abel MH, Kelly RW (1979): Differential production of prostaglandins within the human uterus. Prostaglandins, 18, 821-828.
- Ali M, Buhimschi I, Chwalisz K, Garfield RE (1997): Changes in expression of the nitric oxide synthase isoforms in rat uterus and cervix during pregnancy and parturition. Mol Hum Reprod, 3, 995-1003.
- Arslan A, Zingg HH (1996): Regulation of COX-2 gene expression in rat uterus in vivo and in vitro. Prostaglandins, 52, 463-481.
- Bansal RK, Goldsmith PC, He Y, Zaloudek CJ, Ecker JL, Riemer RK (1997): A decline in myometrial nitric oxide synthase expression is associated with labor and delivery. J Clin Invest, 99, 2502-2508.
- Buhimschi I, Ali M, Jain V, Chwalisz K, Garfield RE (1996): Differential regulation of nitric oxide in the rat uterus and cervix during pregnancy and labour. Hum Reprod, 11, 1755-1766.
- Chwalisz K, Garfield RE (1997): Regulation of the uterus and cervix during pregnancy and labor. Role of progesterone and nitric oxide. Ann N Y Acad Sci, 828, 238-53.
- Dong YL, Gangula PR, Fang L, Yallampalli C (1996): Differential expression of cyclooxygenase-1 and -2 proteins in rat uterus and cervix during the estrous cycle, pregnancy, labor and in myometrial cells. Prostaglandins, 52, 13-34.
- Dong YL, Gangula PRR, Yallampalli C (1996): Nitric oxide synthase isoforms in the rat uterus: differential regulation during pregnancy and labor. J Reprod Fertil, 107, 249-254.
- Fittkow CT, Shi SQ, Bytautiene E, Olson G, Saade GR, Garfield RE (2001): Changes in light-induced fluorescence of cervical collagen in guinea pigs during gestation and after sodium nitroprusside treatment. J Perinat Med, 29, 535-543.
- Ghabour MS, Eis AL, Brockman DE, Pollock JS, Myatt L (1995): Immunohistochemical characterization of placental nitric oxide synthase expression in preeclampsia. Am J Obstet Gynecol, 173, 687-694.
- Gibb W (1998): The role of prostaglandins in human parturition. Ann Med, 30, 235-241.
- Keirse MJ (1993): Prostaglandins in preinduction cervical ripening. Meta-analysis of worldwide clinical experience. J Reprod Med, 38, 89-100.
- Knowles RG, Moncada S (1994): Nitric oxide synthases in mammals. Biochem J, 298, 249-258.
- Ledingham MA, Thomson AJ, Young A, Macara LM, Greer IA, Norman JE (2000): Changes in the expression of nitric oxide synthase in the human uterine cervix during pregnancy and parturition. Mol Hum Reprod, 6, 1041- 1048.
- Marx SG, Wentz MJ, Mackay LB, Schlembach D, Maul H, Fittkow C, Given R, Vedernikov Y, Saade GR, Garfield RE (2006): Effects of progesterone on iNOS, COX-2, and collagen expression in the cervix. J Histochem Cytochem, 54, 623-639.
- Mitchell MD, Romero RJ, Edwin SS, Trautman MS (1995): Prostaglandins and parturition. Reprod Fertil Dev, 7, 623-632.
- Moncada SR, Palmer MG, Higgs EA (1995): Nitric oxide: physiology, pathophysiology and pharmacology. Pharmacol Rev, 43, 109-142.
- Natuzzi ES, Ursell PC, Harrison M, Buscher C, Riemer RK (1993): Nitric oxide synthase activity in the pregnant uterus decreases at parturition. Biochem Biophys Res Commun, 194, 1-8.
- Okawa T, Asano K, Takahashi H, Hashimoto S, Anbe H, Sato A, Vedernikov YP, Saade GR, Garfield RE (2004): Expression of inducible nitric oxide synthase messenger RNA, but not guanylate cyclase messenger RNA, depends on gestational age in rat myometrium. Gynecol Endocrinol, 19, 146-151.
- Olson DM, Mijovic JE, Sadowsky DW (1995): Control of human parturition. Semin Perinatol, 19, 52-63.
- Sladek SM, Regenstein AC, Lykins D, Roberts JM (1993): Nitric oxide synthase activity in pregnant rabbit uterus decreases on the last day of pregnancy. Am J Obstet Gynecol, 169, 1285-1291.
- Smith WL, DeWitt DL (1996): Prostaglandin endoperoxide H synthases-1 and -2. Adv Immunol, 62, 167-215.
- Smith WL, Garavito RM, DeWitt DL (1996): Prostaglandin endoperoxide H synthases (cyclooxygenases)-1 and -2. J Biol Chem, 271, 33157- 33160.
- Swaisgood CM, Zu HX, Perkins DJ, Wu S, Garver CL, Zimmerman PD, Iams JD, Kniss DA (1997): Coordinate expression of inducible nitric oxide synthase and cyclooxygenase-2 genes in uterine tissues of endotoxin- treated pregnant mice. Am J Obstet Gynecol, 177, 1253- 1262.
- Tetsuka T, Daphna-Iken D, Srivastava SK, Baier LD, DuMaine J, Morrison AR (1994): Cross-talk between cyclooxygenase and nitric oxide pathways: prostaglandin E2 negatively modulates induction of nitric oxide synthase by interleukin 1. Proc Natl Acad Sci USA, 91, 12168- 12172.
- Thomson AJ, Telfer JF, Kohnen G, Young A, Cameron IT, Greer IA, Norman JE (1997): Nitric oxide synthase activity and localization do not change in uterus and placenta during human parturition. Hum Reprod, 12, 2546-2552.
- Tschugguel W, Schneeberger C, Lass H, Stonek F, Zaghlula MB, Czerwenka K, Schatten C, Kaider A, Husslein P, Huber JC (1999): Human cervical ripening is associated with an increase in cervical inducible nitric oxide synthase expression. Biol Reprod, 60, 1367-1372.
- Wu WX, Ma XH, Nathanielsz PW (1999): Tissue- specific ontogenic expression of prostaglandin H synthase 2 in the ovine myometrium, endometrium, and placenta during late gestation and at spontaneous term labor. Am J Obstet Gynecol, 181, 1512-1519.
- Yang ZM, Das SK, Wang J, Sugimoto Y, Ichikawa A, Dey SK (1997): Potential sites of prostaglandin actions in the periimplantation mouse uterus: differential expression and regulation of prostaglandin receptor genes. Biology of Reproduction, 56, 368-379.
- Zhang H, Chen X, Teng X, Snead C, Catravas JD (1998): Molecular cloning and analysis of the rat inducible nitric oxide synthase gene promoter in aortic smooth muscle cells. Biochem Pharmacol, 55, 1873-1880.
- Zuo J, Lei ZM, Rao CV, Pietrantoni M, Cook VD (1994): Differential cyclooxygenase-1 and -2 gene expression in human myometria from preterm and term deliveries. J Clin Endocrinol Metab, 79, 894-899.
- Pfaffl MW, Horgan GW, Dempfle L (2002): Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Res, 30, 36.
Details
| Primary Language | English |
|---|---|
| Subjects | Veterinary Surgery |
| Other ID | JA87AC53PF |
| Journal Section | Research Article |
| Authors | |
| Publication Date | December 1, 2012 |
| Published in Issue | Year 2012Volume: 59 Issue: 4 |