Hazel H. Szeto

Mitochondria-Targeted Peptide Accelerates ATP Recovery and Reduces Ischemic Kidney Injury

The burst of reactive oxygen species (ROS) during reperfusion of ischemic tissues can trigger the opening of the mitochondrial permeability transition (MPT) pore, resulting in mitochondrial depolarization, decreased ATP synthesis, and increased ROS production. Rapid recovery of ATP upon reperfusion is essential for survival of tubular cells, and inhibition of oxidative damage can limit inflammation. SS-31 is a mitochondria-targeted tetrapeptide that can scavenge mitochondrial ROS and inhibit MPT, suggesting that it may protect against ischemic renal injury. Here, in a rat model of ischemia-reperfusion (IR) injury, treatment with SS-31 protected mitochondrial structure and respiration during early reperfusion, accelerated recovery of ATP, reduced apoptosis and necrosis of tubular cells, and abrogated tubular dysfunction. In addition, SS-31 reduced medullary vascular congestion, decreased IR-mediated oxidative stress and the inflammatory response, and accelerated the proliferation of surviving tubular cells as early as 1 day after reperfusion. In summary, these results support MPT as an upstream target for pharmacologic intervention in IR injury and support early protection of mitochondrial function as a therapeutic maneuver to prevent tubular apoptosis and necrosis, reduce oxidative stress, and reduce inflammation. SS-31 holds promise for the prevention and treatment of acute kidney injury.

Umbilical blood flow response to embolization of the uterine circulation

The uterine and umbilical blood flow response to uteroplacental insufficiency, produced by microsphere embolization of the uteroplacental circulation, was examined by means of electromagnetic flow transducers in 12 pregnant ewes. No acute changes were observed in either circulation immediately after embolization of the uterine vasculature. Significant morphometric fetal growth retardation, in terms of weight, length, and ponderal index, was not associated with early changes in fetal pH, PO2, or PCO2. However, the incremental increases in both flows that were observed in control animals did not occur either during or after embolization because of an increase in vascular resistance. During and after embolization the incremental increase in uterine blood flow was blunted or decreased in most, but not all, animals. A rapid, progressive, and persistent decrease in umbilical flow occurred in the growth-retarded group. The significance of these findings relative to intrauterine growth is discussed.

A comparison of morphine and methadone disposition in the maternal-fetal unit.

The extent of fetal exposure to a drug is a function of placental and fetal clearances of the drug. We have compared the placental and fetal clearances of morphine and methadone under steady-state conditions in the chronic pregnant ewe preparation. Morphine and methadone were infused at constant rates to the mother and fetus, and steady-state maternal and fetal plasma concentrations of the drugs were determined by radioimmunoassay. At steady state after maternal infusion of the drugs, the maternal: fetal free drug concentration ratios were 7.6 +/- 0.6 for morphine and 2.9 +/- 0.6 for methadone. The clearance of morphine from mother to fetus (24.9 +/- 3.0 ml/min) was fifteenfold less than that of methadone (390.3 +/- 92.8 ml/min). Fetal nonplacental clearance of methadone (381 +/- 89.1 ml/min) was higher than that of morphine (125.6 +/- 15.3 ml/min). Fetal clearance accounted for 67.4 +/- 3.9% of total morphine clearance from the fetal compartment, but only 42.8 +/- 4.9% of total methadone clearance. Fetal clearance of methadone increases with age in the early third trimester. The lower extent of fetal exposure to morphine compared to methadone is due primarily to a slower clearance of morphine across the placenta from mother to fetus rather than a more rapid clearance of morphine by the fetus.

Fetal metabolic response to experimental placental vascular damage

The fetal metabolic response to repetitive placental damage, produced by microsphere embolization of the uteroplacental circulation, was examined longitudinally in eight singleton fetal lambs and compared with similar data from seven controls. Significant morphometric fetal growth retardation was associated with an abrupt cessation of the normal increases in oxygen, glucose, and lactate uptake observed in the control animals. However, fetal blood levels and umbilical substrate quotients did not change. At the time of sacrifice, fetal oxidative metabolic rate was reduced significantly. The significance of these findings relative to intact fetal survival is discussed.

Physiologic variability and fetal electrocortical activity

The relationship between fetal electrocortical activity and multiple fetal parameters was examined in chronically instrumented fetal lambs. Consistent small but significant changes in fetal heart rate, mean arterial pressure, umbilical blood flow, and the cerebral uptake of glucose relative to oxygen were observed as fetal electrocortical activity cyclically shifted between high-voltage slow activity and low-voltage fast activity. Cardiovascular parameters were higher and the cerebral uptake of glucose relative to oxygen was lower during high-voltage slow activity than during low-voltage fast activity. The implication of these observations is discussed.

Disposition of methadone in the ovine maternal-fetal unit.

Abstract The distribution of methadone between mother and fetus after a single dose and at steady state was determined using the chronic pregnant ewe preparation. Chronic indwelling catheters were placed in the maternal aorta and vena cava, umbilical vein and fetal aorta. Following a single i.v. dose (0.5 mg/kg) to the mother, methadone was rapidly distributed to the fetus, with peak concentration in the umbilical vein occurring within two min. An umbilical venous-arterial gradient existed for 10–15 min after drug administration, indicating uptake of methadone by fetal tissues. Methadone concentration in the fetus was 2–5 times lower than those in the mother even in the post-distribution phase. The terminal half-life of methadone in 4 animals was 57±7.6 (S.E.) min in the mother, and 58.5±10.0 (S.E.) min in the fetus. When methadone was infused at a constant rate to the mother (0.01 mg/kg/min), steady state was achieved in both mother and fetus by 4–5 hrs. In 5 animals, maternal steady state was found to be 203±18.8 (S.E.) ng/ml, and fetal steady state was found to be 29.7±2.9 (S.E.) ng/ml. These studies show that methadone is rapidly distributed to the fetus, but fetal concentration remain lower than maternal concentration at all times.

Baroreflex-mediated bradycardia but not tachycardia is blunted peripherally by intravenous μ-opioid agonists

OBJECTIVEnWe sought to test the hypothesis that an intravenous dose of H-Tyr-D-Arg-Phe-Lys-NH2, a highly mu-receptor selective opioid peptide, suppresses baroreflex sensitivity through a peripheral mechanism.nnnSTUDY DESIGNnA transient change in mean arterial pressure was produced in chronically instrumented pregnant ewes by norepinephrine or sodium nitroprusside in the absence or in the presence of H-Tyr-D-Arg-Phe-Lys-NH2, a highly mu-selective opioid peptide. In some studies naloxone methiodide, a peripheral opioid antagonist, was infused starting 60 minutes before the administration of H-Tyr-D-Arg-Phe-Lys-NH2 and maintained for a total of 90 minutes. Linear plots were obtained when the changes in mean arterial pressure during the pressure rise were plotted against the changes in heart rate and the sensitivity of the baroreflex was derived as the slope of the linear regression line.nnnRESULTSnWe observed (1) lower baroreflex sensitivity after H-Tyr-D-Arg-Phe-Lys-NH2 administration with a hypertensive stimulus; (2) unchanged baroreflex sensitivity after H-Tyr-D-Arg-Phe-Lys-NH2 administration with a hypotensive stimulus; and (3) unchanged baroreflex sensitivity after H-Tyr-D-Arg-Phe-Lys-NH2 administration with a hypertensive stimulus in the presence of naloxone methiodide.nnnCONCLUSIONnH-Tyr-D-Arg-Phe-Lys-NH2 suppresses the hypertensive but not the hypotensive arm of the baroreflex through peripheral opioid receptors. These results suggest that mu-opioid receptors are present in the vagus nerves and that the activation of these opioid receptors inhibits reflex bradycardia in pregnant sheep.

A Peripheral Site of Action for the Attenuation of Baroreflex-Mediated Bradycardia by Intravenous μ-Opioid Agonists

We previously reported that i.v. DAMGO (Tyr-D-Ala-Gly-NMePhe-Gly-ol), a selective mu-opioid agonist, causes an increase in blood pressure with no change in heart rate in unanesthetized sheep and subsequently demonstrated that DAMGO attenuates baroreflex-mediated bradycardia. To determine the site and mechanism by which mu-agonists inhibit baroreflex sensitivity, we have carried out further investigations by using DAMGO and another mu-agonist, DALDA (Tyr-D-Arg-Phe-Lys-NH2). The bradycardic response to norepinephrine (NE) was significantly blunted after i.v. DAMGO or DALDA in both nonpregnant and pregnant sheep. In contrast, the tachycardic response to sodium nitroprusside (SNP) remained unchanged in the presence of DAMGO or DALDA. In view of the highly restricted distribution of DALDA across the blood-brain barrier (BBB), we hypothesized that the blunting of reflex-mediated bradycardia by mu-opioid agonists can occur peripherally. Pretreatment with the quaternary opioid antagonist, naloxone methiodide (NM), completely blocked the attenuation of baroreflex sensitivity by DAMGO and DALDA in both nonpregnant and pregnant animals. These data suggest that in addition to central mechanisms, mu-opioid agonists can inhibit baroreflex sensitivity at a peripheral site, most likely by inhibiting vagal influence on heart-rate control rather than by acting directly at baroreceptors.

Pharmacologic Approaches to Improve Mitochondrial Function in AKI and CKD

AKI is associated with high morbidity and mortality, and it predisposes to the development and progression of CKD. Novel strategies that minimize AKI and halt the progression of CKD are urgently needed. Normal kidney function involves numerous different cell types, such as tubular epithelial cells, endothelial cells, and podocytes, working in concert. This delicate balance involves many energy-intensive processes. Fatty acids are the preferred energy substrates for the kidney, and defects in fatty acid oxidation and mitochondrial dysfunction are universally involved in diverse causes of AKI and CKD. This review provides an overview of ATP production and energy demands in the kidney and summarizes preclinical and clinical evidence of mitochondrial dysfunction in AKI and CKD. New therapeutic strategies targeting mitochondria protection and cellular bioenergetics are presented, with emphasis on those that have been evaluated in animal models of AKI and CKD. Targeting mitochondrial function and cellular bioenergetics upstream of cellular damage may offer advantages compared with targeting downstream inflammatory and fibrosis processes.

Resensitization of Blood Pressure Response to μ-Opioid Peptide Agonists After Acute Desensitization

IV administration of &mgr;-opioid peptide agonists (DAMGO, DALDA, and [Dmt1]DALDA) results in a transient, naloxone-sensitive, increase in blood pressure in awake sheep. Despite significant differences in pharmacokinetics, these blood pressure responses all last <15 min. The lack of correlation between half-life and duration of action suggested rapid desensitization. When a second dose of the same agonist was repeated 30 min later, the response was completely abolished. An increase in blood pressure and rapid desensitization was also observed with the &kgr;-opioid agonist (U50488H), whereas &dgr;-agonists (DPDPE and DELT) had no effect on blood pressure. The response to DAMGO was abolished after prior exposure to DAMGO or DALDA, but there was no evidence of cross-desensitization between &mgr; and &dgr;, or &mgr; and &kgr;, opioid agonists. Full resensitization of the blood pressure response occurred by 4 h for DAMGO (t1/2 = 15 min) and by 48 h for [Dmt1]DALDA (t1/2 = 1.8 h). These data support our hypothesis that the transient nature of the blood pressure response to &mgr;-opioid agonists is caused by rapid desensitization and suggest that the rate of resensitization is dependent on the pharmacokinetics of the agonist.