endothelin-1 and Abnormalities--Multiple

To investigate the expression of endothelin-1 (ET-1) and adrenomedullin (ADM) in the renal pelvis, stenotic ureteropelvic junction, and ureter of 20 male Wistar rats with congenital unilateral ureteropelvic junction obstruction; the normal contralateral kidneys served as controls. The molecular pathophysiology of congenital ureteropelvic junction obstruction is still unclear. The implication of altered peptidergic innervation is under discussion. Our study group has recently been able to demonstrate a significant increase in ET-1 and a significant decrease in ADM in prestenotic and stenotic tissue, but not in the remainder of the ureter, compared with controls.. Twenty animals were killed, and samples of the renal pelvis, ureteropelvic junction, upper ureter, middle part of the ureter, and lower ureter were immediately snap-frozen and stored in liquid nitrogen. Total RNA was extracted, and subsequently 1 microg of RNA was reversely transcribed. mRNA expression of ET-1 and ADM was determined semiquantitatively using on-line polymerase chain reaction. The expression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was determined to relate the specific mRNA expression to the expression of a housekeeping gene.. We found a significant increase in the expression of ET-1 in the obstructed junctions related to GAPDH (P <0.001). The expression of ADM, however, revealed no statistically significant differences. No differences at all could be detected in the tissue samples from the rest of the ureter.. Alterations in the local production of peptidergic neurotransmitters, especially ET-1, may contribute to the molecular pathogenesis of ureteropelvic junction obstruction. Results previously obtained in the stenotic tissue from children were confirmed in the stenotic tissue from the rat model. We hypothesize that the alterations are disease-, but not age-specific.

Topics: Abnormalities, Multiple; Adrenomedullin; Animals; Atrophy; Computer Systems; Constriction, Pathologic; Disease Models, Animal; Endothelin-1; Gene Expression Profiling; Hydronephrosis; Kidney Pelvis; Male; Peptides; Polymerase Chain Reaction; Rats; Rats, Mutant Strains; Rats, Wistar; RNA, Messenger; Ureter; Ureteral Obstruction

Endothelin-1 (ET-1), a potent vasoconstrictor peptide and modulator of vasoactive substances such as prostanoids and nitric oxide (NO), plays an important role during embryo and fetal development. In this work, ET-1, nitrate and nitrite, and prostaglandin E2 (PGE2) levels in embryos from control and neonatal streptozotocin-induced (n-stz) diabetic rats were assessed, and the modulatory pathways regulating the generation of these vasoactive agents investigated. Endothelin-1 concentrations were found to be increased in embryos from n-stz diabetic rats when compared with controls. Additions of spermine NONOate, a nitric oxide donor, enhanced ET-1 levels in embryos from both control and n-stz diabetic rats, whereas N(G)-monomethyl-L-arginine, a nitric oxide inhibitor, diminished embryonic ET-1 content. Thus, enhanced ET-1 levels in the embryos from n-stz diabetic rats may be related to the elevated NO levels found in those embryos. Additions of ET-1 or bosentan (an endothelin A and endothelin B receptor antagonist), did not alter PGE2 generation in embryos from either control or n-stz diabetic rats. Endothelin-1 additions diminished nitrate and nitrite levels in embryos from both control and n-stz diabetic rats, whereas bosentan stimulated nitrate and nitrite generation in those embryos. In the present work, it was found that ET-1 levels were enhanced in embryos from n-stz diabetic rats, probably as a result of NO overproduction, an alteration which may be related to embryonic abnormalities and growth delay. Endothelin-1 has been shown to be a negative modulator of embryonic NO levels, a mechanism likely to be important during development. Endothelin-1 may prevent damage induced by NO overproduction in embryos from n-stz diabetic rats.

Topics: Abnormalities, Multiple; Animals; Animals, Newborn; Case-Control Studies; Diabetes Mellitus, Experimental; Dinoprostone; Embryo, Mammalian; Endothelin-1; Female; Nitrates; Nitric Oxide; Nitric Oxide Donors; Nitrites; omega-N-Methylarginine; Pregnancy; Pregnancy in Diabetics; Proteins; Rats; Rats, Wistar; Spermine; Streptozocin