title: Mechanisms underlying the contractile response to endothelin-1 in the rat renal artery.
abstract: We assessed the functional response and the mechanisms following receptor stimulation of endothelin-1 (ET-1) in the rat renal artery. In this study, isometric tension was recorded in renal artery rings without endothelium. Cumulative application of ET-1 from 0.1 to 100 nmol/l induced a sustained concentration-dependent contraction in the renal artery. Submaximal contraction induced by 10 nmol/l ET-1 in 2.5 mmol/l Ca(2+) and in the absence of inhibitors was used as control response (100%). The relative contribution of different sources of Ca(2+) in ET-1-induced contraction was evaluated. The contractile response to 10 nmol/l ET-1 in 2.5 mmol/l Ca(2+ )(1.2 +/- 0.2 g) was significantly inhibited either in Ca(2+)-free solution containing 100 micromol/l ethylene glycol bis-(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (0.6 +/- 0.1 g) or after depletion of intracellular Ca(2+) stores (0.62 +/- 0.05 g). The contribution of phospholipase C and protein kinase C was evaluated by using their inhibitors 2-nitro-4-carboxyphenyl N,N-diphenylcarbamate (NCDC) and [1-(5-isoquinolinesulfonyl)-2-methylpiperazine] (H-7), respectively. The contractile response to 10 nmol/l ET-1 was inhibited by 10 micromol/l NCDC (to 80 +/- 6%) and 30 micromol/l H-7 (to 76.6 +/- 6.5%). We found that 1 micromol/l nifedipine inhibited the ET-1-induced contraction (to 48.7 +/- 6.9%), indicating the contribution of Ca(2+) influx through voltage-gated L-type Ca(2+) channels to this response. Further, the inhibitory effect of nifedipine was to a greater extent as compared with NCDC or H-7. Additive inhibition of ET-1-induced contraction was not observed in the presence of both nifedipine and NCDC. We also evaluated the role of the ionic transport system in the ET-1-induced response by using 20 nmol/l 5-(N-ethyl-N-isopropyl)-amiloride (EIPA), an inhibitor of Na(+)-H(+) exchange, or 100 micromol/l ouabain, an inhibitor of Na(+)-K(+)-ATPase. The response to ET-1 was decreased by both EIPA (to 61.6 +/- 8.4%) and ouabain (to 62.1 +/- 8.6%). The contribution of Na(+)-Ca(2+) exchange to ouabain action was tested using the inhibitor dimethyl amiloride HCl (10 micromol/l). The decrease in ET-1-induced contraction by the combination of ouabain and dimethyl amiloride HCl was similar to that observed with ouabain alone. In view of these observations, both extra- and intracellular sources of Ca(2+) contribute to the contractile response induced by ET-1 in the renal artery. Our findings also revealed the importance of Ca(2+) influx through voltage-gated L-type Ca(2+) channels in mediating contraction to ET-1 in the renal artery, whereas a minor role of phospholipase C and protein kinase C was observed. Na(+)-H(+) exchange and Na(+)-K(+)-ATPase also play a role in the ET-1-induced contraction in renal artery. Moreover, the contribution of Na(+)-K(+)-ATPase in ET-1 contraction is not an Na(+)-Ca(2+) exchange-related process.
authors.affiliation: Laboratory of Pharmacology, Faculty of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto, Brazil.
authors.forename: Glaucia Elena
authors.initials: GE
authors.lastname: Callera
authors.forename: Lusiane Maria
authors.initials: LM
authors.lastname: Bendhack
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D015220:Calcium Channels / Q000378:metabolism; D019332:Endothelin-1 / Q000494:pharmacology*; D066298:In Vitro Techniques; D007668:Kidney / Q000098:blood supply; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000187:drug effects* / Q000502:physiology*; D051381:Rats; D017208:Rats, Wistar; D012077:Renal Artery / Q000187:drug effects* / Q000502:physiology; D014661:Vasoconstriction / Q000187:drug effects*
chemical_list: D015220:Calcium Channels; D019332:Endothelin-1; D002118:Calcium