Biochemical and molecular endothelin responses to morphine sulfate infusion in conscious newborn piglets

The biochemical and molecular endothelin-1 (ET-1) responses to high dose morphine sulfate infusion were studied in conscious newborn piglets (n = 6) that received a loading dose of 100 micrograms/kg over 5 min followed by a continuous i.v. infusion dose of 100 micrograms.kg-1.h-1 for 4 h. The control group (n = 6) received equivalent volume loading and infusion doses of 5% dextrose. Blood samples were drawn serially from the femoral artery and sagittal sinus vein before (0), during (30 min, 1, 2, 3, and 4 h), and post (1 and 2 h) infusion. Five micrograms of total RNA obtained from brainstem tissue homogenates was analyzed by reverse transcriptase--polymerase chain reaction (RT-PCR). The amounts of mRNA encoding ET-1, and endothelin receptor subtypes ETA and ETB, were semiquantitated using densitometric scanning. Morphine infusion resulted in elevated respiratory rate and mean arterial blood pressure, with no effect on arterial pH, Po2, and O2 saturation. Compared with the control group, morphine induced significant elevations in plasma ET-1 levels following the bolus dose (systemic: 13.2 +/- 3.6 vs. 8.6 +/- 2.2 pg/mL, p < 0.05; sagittal sinus vein: 13.7 +/- 3.4 vs. 8.2 +/- 0.9 pg/mL, p < 0.01). These effects lasted up to 2 h after discontinuation of morphine infusion (systemic: 14.5 +/- 3.4 to 18.7 +/- 5.7 pg/mL vs. 7.5 +/- 0.8 to 9.4 +/- 3.2 pg/mL, p < 0.05 to p < 0.01; sagittal sinus vein: 14.8 +/- 2.7 to 17.6 +/- 2.8 pg/mL vs. 7.5 +/- 1.4 to 9.4 +/- 3.4 pg/mL, p < 0.05 to p < 0.01). The RT-PCR assay showed a twofold (p < 0.02) upregulation in ET-1 and a threefold (p < 0.007) upregulation in ETA receptor mRNA expression in the brainstem of morphine-treated animals. In contrast, there was a threefold (p < 0.0001) downregulation of the ETB receptor mRNA expression. The rapid and sustained elevations in systemic arterial and sagittal sinus venous ET-1 levels suggest a role for ET-1 in the morphine-induced excitatory responses observed in newborn piglets. Upregulation of ETA receptors and downregulation of ETB receptors in the brainstem with high doses of morphine may indicate possible effects on cerebral vascular tone.