Search

Cystic fibrosis (CF) pigs develop disease with features remarkably similar to those in people with CF, including exocrine pancreatic destruction, focal biliary cirrhosis, micro-gallbladder, vas deferens loss, airway disease, and [...]

Exercise increases serum opioid levels and improves cardiovascular health. Here we tested the hypothesis that opioids contribute to the acute cardioprotective effects of exercise using a rat model of exercise-induced cardioprotection. For the standard protocol, rats were randomized to 4 days of treadmill training and 1 day of vigorous exercise (day 5), or to a sham exercise control group. On day 6, animals were killed, and global myocardial ischemic tolerance was assessed on a modified Langendorff apparatus. Twenty minutes of ischemia followed by 3 h of reperfusion resulted in a mean infarct size of 42 [+ or -] 4% in hearts from sham exercise controls and 21 [+ or -] 3% (P < 0.001) in the exercised group. The cardioprotective effects of exercise were gone by 5 days after the final exercise period. To determine the role of opioid receptors in exercise-induced cardioprotection, rats were exercised according to the standard protocol; however, just before exercise on days 4 and 5, rats were injected subcutaneously with 10 mg/kg of the opioid receptor antagonist naltrexone. Similar injections were performed in the sham exercise control group. Naltrexone had no significant effect on baseline myocardial ischemic tolerance in controls (infarct size 43 [+ or -] 4%). In contrast, naltrexone treatment completely blocked the cardioprotective effect of exercise (infarct size 40 [+ or -] 5%). Exercise was also associated with an early increase in myocardial mRNA levels for several opioid system genes and with sustained changes in a number of genes that regulate inflammation and apoptosis. These findings demonstrate that the acute cardioprotective effects of exercise are mediated, at least in part, through opioid receptor-dependent mechanisms that may include changes in gene expression. exercise; myocardial ischemic tolerance; enkephalins; opioid receptors

Diabetes is a common and significant co-morbidity in cystic fibrosis (CF). The pathogenesis of cystic fibrosis related diabetes (CFRD) is incompletely understood. Because exocrine pancreatic disease is similar between humans and pigs with CF, the CF pig model has the potential to contribute significantly to the understanding of CFRD pathogenesis. We determined the structure of the endocrine pancreas in fetal, newborn and older CF and non-CF pigs and assessed endocrine pancreas function by intravenous glucose tolerance test (IV-GTT). In fetal pigs, pancreatic insulin and glucagon density was similar between CF and non-CF. In newborn and older pigs, the insulin and glucagon density was unchanged between CF and non-CF per total pancreatic area, but increased per remnant lobular tissue in CF reflecting exocrine pancreatic loss. Although fasting glucose levels were not different between CF and non-CF newborns, CF newborns demonstrated impaired glucose tolerance and increased glucose area under the curve during IV-GTT. Second phase insulin secretion responsiveness was impaired in CF newborn pigs and significantly lower than that observed in non-CF newborns. Older CF pigs had elevated random blood glucose levels compared with non-CF. In summary, glycaemic abnormalities and insulin secretion defects were present in newborn CF pigs and spontaneous hyperglycaemia developed over time. Functional changes in CF pig pancreas were not associated with a decline in islet cell mass. Our results suggest that functional islet abnormalities, independent of structural islet loss, contribute to the early pathogenesis of CFRD. [ABSTRACT FROM AUTHOR]

Enkephalins are opioid peptides that act as neurotransmitters and modulators of cardiac function. Plasma enkephalin levels increase in response to ischemia and exercise. The level of bioactive peptides in blood is regulated at many levels, including inactivation by plasma or cell-associated peptidases. Previous studies showed that Met-enkephalin (ME) and Leu-enkephalin (LE) bound to plasma proteins, and that binding to these proteins protected LE from proteolysis. We found that Met-enkephalin-Arg-Phe (MEAP), and Met-enkephalin-Arg-Gly-Leu (MEAGL) also bound to human serum proteins; however, this binding was of significantly higher affinity that that of ME and LE. In addition, the enkephalin binding capacity of plasma was considerably greater than that of serum from the same donor, suggesting that enkephalins avidly bind to proteins that are present in plasma but not in serum. Finally, we observed a unique degradation pattern when MEAP was hydrolyzed by plasma carboxypeptidases, as compared to ME, LE and MEAGL. Together, our data suggest complex mechanisms by which enkephalin-plasma interactions may regulate systemic enkephalin bioactivity. [ABSTRACT FROM AUTHOR]

Ischemia-reperfusion injury contributes to the pathophysiology of numerous diseases. Ischemic preconditioning increases tissue resistance to ischemia-reperfusion injury. Among the mediators of ischemic preconditioning are opioid peptides called enkephalins. We found that numerous non-neuronal tissues express the enkephalin precursor protein proenkephalin. Additionally, using isolated heart, intestine, and skeletal muscle, we found a tissue-specific pattern of enkephalin release. We also found that ischemia increased cardiac enkephalin release, and that exercise altered the efficiency of enkephalin processing and release. In addition to acting as potential sources for enkephalin release, heart, intestine, and skeletal muscle all express mRNAs for opioid receptors, suggesting that these tissues are also targets for opioid-mediated effects. Consistent with this conclusion, purified enkephalins induced opioid-dependent decreases in intestinal contractility. Taken together, our data suggest that skeletal muscle, heart, and intestine may release enkephalins that may in turn mediate autocrine, paracrine, and systemic effects, including ischemic preconditiong. [ABSTRACT FROM AUTHOR]

Ischemic heart disease is a major contributor to death worldwide. Ischemic preconditioning (IPC) is a natural process that protects the heart from ischemic injury. The goal of our laboratory is to determine the mechanisms by which mediators of IPC exert their protective effects. Among the mediators of IPC are small opioid peptides called enkephalins. We found that numerous tissues, including skeletal muscle, express the precursor protein proenkephalin (PENK), and release enkephalins in a tissue-specific manner. We also found that exercise, which is potently cardioprotective, alters PENK levels in both heart and skeletal muscle, but not in intestine. Moreover, ischemia-induced enkephalin release is higher in hearts from exercised animals, as compared to control animals. Together, our data support a model wherein numerous non-neuronal tissues release enkephalins in response to ischemia and ischemic-like stresses (e.g., exercise). These enkephalins may then act locally or systemically to increase ischemic tolerance of the heart and other tissues. Our findings further suggest that enhancing or mimicking the protective effects of enkephalins may benefit patients with ischemic disease. [ABSTRACT FROM AUTHOR]

Background: Acute myocardial ischemia is an important cause of morbidity and mortality worldwide. The heart and other organs can be rendered more resistant to the deleterious effects of ischemia through a variety of preconditioning strategies, including treadmill exercise and brief ischemia of skeletal muscle. Some of the beneficial effects of these preconditioning strategies appear to be mediated by as-of-yet unidentified hormonal opioids., Objectives: To test the hypothesis that endogenous opioids of the enkephalin class are capable of improving ischemic tolerance and acting in a hormonal manner., Methods: In phase one of the investigation, the authors assessed the cardioprotective potential of all four known enkephalins. This was achieved by subjecting isolated buffer-perfused rabbit hearts to a 25-minute period of test ischemia and two hours of reperfusion (protocol 1) after receiving treatment with either saline vehicle (controls) or increasing concentrations of purified enkephalins. On the basis of results from these initial studies, the authors performed additional experiments (protocol 2) to determine whether Met5-enkephalin-Arg6-Phe7 (MEAP) could be absorbed from skeletal muscle and exert a cardioprotective effect. Specifically, MEAP or vehicle (controls) was given intramuscularly 24 hours before the hearts were harvested. A similar assessment of ischemic tolerance as described in protocol 1 was then performed. Postischemic myocardial viability (infarct size) was assessed in all cases by triphenyltetrazolium chloride (TTC) staining. Hemodynamic parameters and infarct sizes for concentration-dependence studies were compared by two-way analysis of variance, and infarct sizes from protocol 2 studies were compared by using Student's t-test (significance set at p < or = 0.05)., Results: Mean infarct size in control hearts (+/- SEM) was 33% (+/- 4%) and 36% (+/- 6%) for protocol 1 and 2, respectively. Of the four enkephalins tested in protocol 1, only MEAP treatment showed a tendency toward cardioprotection. Interestingly, an alternative enkephalin, methionine5-enkephalin-Arg6-Gly7-Leu8, tended to exert an injurious effect. In protocol 2, MEAP treatment 24 hours before ischemia significantly reduced infarct size (14% +/- 4%) compared with controls, suggesting that it can be released from muscle and exert a distant cardioprotective effect., Conclusions: When given either directly to the heart or absorbed from a distant tissue, MEAP induces cardioprotection, supporting the hypothesis that it can act as a hormonal modulator of ischemic tolerance.