Intermittent High Altitude Hypoxia Research Articles

ZFP580, a novel zinc-finger transcription factor, is involved in cardioprotection of intermittent high-altitude hypoxia against myocardial ischemia-reperfusion injury.

BackgroundZFP580 is a novel C2H2 type zinc-finger transcription factor recently identified by our laboratory. We previously showed that ZFP580 may be involved in cell survival and growth. The aim of this study was to elucidate whether ZFP580 is involved in the cardioprotective effects of intermittent high-altitude (IHA) hypoxia against myocardial ischemia-reperfusion (I/R) injury.Methods and ResultsAfter rats were subjected to myocardial ischemia for 30 min followed by reperfusion, ZFP580 expression in the left ventricle was measured. ZFP580 protein expression was found to be up-regulated within 1 h and decreased at 2 h after reperfusion. Comparing normoxic and IHA hypoxia-adapted rats (5000 m, 6 h day−1, 6 weeks) following I/R injury (30 min ischemia and 2 h reperfusion), we found that adaptation to IHA hypoxia attenuated infarct size and plasma leakage of lactate dehydrogenase and creatine kinase-MB. In addition, ZFP580 expression in the myocardium was up-regulated by IHA hypoxia. Consistent with this result, ZFP580 expression was found to be significantly increased in cultured H9c2 myocardial cells in the hypoxic preconditioning group compared with those in the control group following simulated I/R injury (3 h simulated ischemic hypoxia and 2 h reoxygenation). To determine the role of ZFP580 in apoptosis, lentivirus-mediated gene transfection was performed in H9c2 cells 72 h prior to simulated I/R exposure. The results showed that ZFP580 overexpression significantly inhibited I/R-induced apoptosis and caspase-3 activation. H9c2 cells were pretreated with or without PD98059, an inhibitor of ERK1/2 phosphorylation, and Western blot results showed that PD98059 (10 µM) markedly suppressed I/R-induced up-regulation of ZFP580 expression.ConclusionsOur findings demonstrate that the cardioprotective effect of IHA hypoxia against I/R injury is mediated via ZFP580, a downstream target of ERK1/2 signaling with anti-apoptotic roles in myocardial cells.

Cardiac Tolerance to Ischemia in Neonatal Spontaneously Hypertensive Rats

Hypertension is the risk factor of serious cardiovascular diseases, such as ischemic heart disease and atherosclerosis. The aim of the present study was to analyze the development of cardiac tolerance to ischemia in neonatal spontaneously hypertensive rats (SHR) and possible protective effect of ischemic preconditioning (IP) or adaptation to intermittent high-altitude hypoxia (IHAH). For this purpose we used 1- and 10-day-old pups of SHR and their normotensive control Wistar Kyoto rats (WKY). Isolated hearts were perfused in the Langendorff mode with Krebs-Henseleit solution at constant pressure, temperature and rate. Cardiac tolerance to ischemia was expressed as a percentage of baseline values of developed force (DF) after global ischemia. IP was induced by three 3-min periods of global ischemia, each separated by 5-min periods of reperfusion. IHAH was simulated in barochamber (8 h/day, 5000 m) from postnatal day 1 to 10. Cardiac tolerance to ischemia in 1-day-old SHR was higher than in WKY. In both strains tolerance decreased after birth, and the difference disappeared. The high cardiac resistance in 1- and 10-day-old SHR and WKY could not be further increased by both IP and adaptation to IHAH. It may be concluded that hearts from newborn SHR are more tolerant to ischemia/reperfusion injury as compared to age-matched WKY; cardiac resistance decreased in both strains during the first ten days, similarly as in Wistar rats.

Calcium/calmodulin-dependent protein kinase II mediates cardioprotection of intermittent hypoxia against ischemic-reperfusion-induced cardiac dysfunction

Intermittent high-altitude (IHA) hypoxia-induced cardioprotection against ischemia-reperfusion (I/R) injury is associated with the preservation of sarcoplasmic reticulum (SR) function. Although Ca(2+)/calmodulin (CaM)-dependent protein kinase II (CaMKII) and phosphatase are known to modulate the function of cardiac SR under physiological conditions, the status of SR CaMKII and phosphatase during I/R in the hearts from IHA hypoxic rats is unknown. In the present study, we determined SR and cytosolic CaMKII activity during preischemia and I/R (30 min/30 min) in perfused hearts from normoxic and IHA hypoxic rats. The left ventricular contractile recovery, SR CaMKII activity as well as phosphorylation of phospholamban at Thr(17), and Ca(2+)/CaM-dependent SR Ca(2+)-uptake activity were depressed in the I/R hearts from normoxic rats, whereas these changes were prevented in the hearts from IHA hypoxic rats. Such beneficial effects of IHA hypoxia were lost by treating the hearts with a specific CaMKII inhibitor, KN-93. I/R also depressed cytosolic CaMKII and SR phosphatase activity, but these alterations remained unchanged in IHA hypoxic group. Furthermore, we found that the autophosphorylation at Thr(287), which confers Ca(2+)/CaM-independent activity, was not altered by I/R in both groups. These findings indicate that preservation of SR CaMKII activity plays an important role in the IHA hypoxia-induced cardioprotection against I/R injury via maintaining SR Ca(2+)-uptake activity.

Abstract 241: Intermittent Hypoxia Protects Cardiomyocytes Against Ischemia/Reperfusion-Induced Ca 2+ Overload and Contraction Suppression via Activation of α 1b -adrenoceptors

We have demonstrated that long-term training to intermittent high altitude hypoxia (IH) protects hearts against ischemia-reperfusion (I/R)-induced Ca 2+ overload and improved postischemic myocardial contractility. This cardioprotection is non-invasive and maintains much longer time compared with that induced by ischemic preconditioning, thereby, it may have great clinical potential. However, its precise signaling pathways are not fully clarified. Here we hypothesized that IH may attenuate I/R-induced cytosolic and mitochondrial Ca 2+ overload via activation of α 1 -adrenoceptor (α 1 -ARs) pathways. To test it, dynamics of cytosolic free Ca 2+ concentration ([Ca 2+ ] c ), mitochondrial [Ca 2+ ] ([Ca 2+ ] m ) and cell contraction were examined during preischemia and I/R in ventricular cardiomyocytes from normoxic and IH rats. IH significantly attenuated I/R-induced [Ca 2+ ] c and [Ca 2+ ] m overload by 63.5 ± 3.6% and 57.0 ± 4.5% at 30 min of reperfusion (R30), respectively. Consistently, I/R-induced depression on Ca 2+ transients and cell shortening was markedly attenuated by IH, with decreases in mitochondrial cytochrome c release and ATP reduction during reperfusion. Meanwhile, the α 1B -AR and α 1D -AR density obviously decreased during ischemia, while α 1A -AR density remained unchanged. IH did not change α 1A -AR density, but increased α 1B -AR density by ~50% and decreased α 1D -AR density further by 30% during I/R. Moreover, IH-induced protections were abolished by α 1B -AR antagonist chloroethylclonidine but not α 1A - or α 1D -AR antagonist. Furthermore, IH-afforded protection was lost when epsilon isoform of protein kinase C (PKCϵ) was inhibited with PKCϵ V1–2 and mitochondrial ATP sensitive potassium (mitoK ATP ) channels with 5-hydroxydecanoate, a downstream signaling pathway and effector of α 1 -ARs. Our findings indicate that α 1B -AR subtype might be one of the important mediators in IH protects myocytes against I/R injury via maintaining intracellular Ca 2+ homeostasis and protecting the ability of mitochondria through activation of the α 1B -ARs and PKCϵ pathway as well as mitoK ATP channel.. by via translocation of PKCϵ and opening of mitoK ATP channels. Mitochondrial function appears to be a determinant of cardioprotection induced by IH.

Intermittent high altitude hypoxia inhibits opening of mitochondrial permeability transition pores against reperfusion injury

The role of mitochondrial permeability transition pore (MPTP) in the mechanism of intermittent high altitude (IHA) hypoxic adaptation is not understood. Therefore, we study whether the protective effect of IHA hypoxia against ischemia–reperfusion injury is accompanied by inhibition of MPTP opening. IHA hypoxia significantly improved the functional recovery of Langendorff hearts on reperfusion and limited infarct size. In isolated myocytes, IHA hypoxia significantly improved the recovery of cell length, lowered ischemia–reperfusion-induced [Ca 2+]c and [Ca 2+]m overloading. Furthermore, IHA hypoxia accelerated [Ca 2+]c decline during reperfusion. Opening the MPTP with atractyloside immediately at reperfusion abolished these cardioprotective effects of IHA hypoxia, but had no appreciable influence on those of normoxic hearts. IHA hypoxia prolongs the time taken to induce MPTP opening and the time taken to induce rigor contracture when myocytes subjected to oxidative stress. The data from isolated mitochondria demonstrated that IHA hypoxia prevented the decrease of ADP/O ratio, the opening of MPTP and the release of cytochrome c from mitochondria at high Ca 2+ concentrations (100 and 200 μM). Inhibition of MPTP opening in the first few minutes of reperfusion accelerated [Ca 2+]c decline and attenuated [Ca 2+]c and [Ca 2+]m overloading, which contributed to the cardioprotection of IHA hypoxic adaptation. Enhancement of the tolerance of mitochondria against Ca 2+ might underlie the protective mechanism of IHA hypoxia.

Role of dual-site phospholamban phosphorylation in intermittent hypoxia-induced cardioprotection against ischemia-reperfusion injury

Cardioprotection by intermittent high-altitude (IHA) hypoxia against ischemia-reperfusion (I/R) injury is associated with Ca(2+) overload reduction. Phospholamban (PLB) phosphorylation relieves cardiac sarcoplasmic reticulum (SR) Ca(2+)-pump ATPase, a critical regulator in intracellular Ca(2+) cycling, from inhibition. To test the hypothesis that IHA hypoxia increases PLB phosphorylation and that such an effect plays a role in cardioprotection, we compared the time-dependent changes in the PLB phosphorylation at Ser(16) (PKA site) and Thr(17) (CaMKII site) in perfused normoxic rat hearts with those in IHA hypoxic rat hearts submitted to 30-min ischemia (I30) followed by 30-min reperfusion (R30). IHA hypoxia improved postischemic contractile recovery, reduced the maximum extent of ischemic contracture, and attenuated I/R-induced depression in Ca(2+)-pump ATPase activity. Although the PLB protein levels remained constant during I/R in both groups, Ser(16) phosphorylation increased at I30 and 1 min of reperfusion (R1) but decreased at R30 in normoxic hearts. IHA hypoxia upregulated the increase further at I30 and R1. Thr(17) phosphorylation decreased at I30, R1, and R30 in normoxic hearts, but IHA hypoxia attenuated the depression at R1 and R30. Moreover, PKA inhibitor H89 abolished IHA hypoxia-induced increase in Ser(16) phosphorylation, Ca(2+)-pump ATPase activity, and the recovery of cardiac performance after ischemia. CaMKII inhibitor KN-93 also abolished the beneficial effects of IHA hypoxia on Thr(17) phosphorylation, Ca(2+)-pump ATPase activity, and the postischemic contractile recovery. These findings indicate that IHA hypoxia mitigates I/R-induced depression in SR Ca(2+)-pump ATPase activity by upregulating dual-site PLB phosphorylation, which may consequently contribute to IHA hypoxia-induced cardioprotection against I/R injury.

Low body weight and cardiac tolerance to ischemia in neonatal rats.

Adaptation to intermittent high altitude hypoxia (IHAH) increases tolerance of the isolated neonatal rat heart to ischemia and potentiates protection induced by ischemic preconditioning. In addition to the protective effect, IHAH significantly reduces growth of the animals. The aim of the present study was, therefore, to find out whether low body weight per se might influence cardiac sensitivity to oxygen deprivation. Low body weight was induced either by IHAH (barochamber, 8 h/day, 5000 m) from postnatal day 1 to 10 (HLBW), or by a higher number of sucklings per mother (14 instead of 8), again from postnatal day 1 to 10 (NLBW). Control animals (8 littermates per mother) were kept under normoxic conditions (Controls). The recovery of developed force following 40 min of global ischemia was measured in isolated hearts from 10-day-old rats by perfusing them in the Langendorff mode with Krebs-Henseleit solution at constant pressure, temperature and rate. Ischemic preconditioning was induced by three 3-min periods of global ischemia, each separated by 5-min periods of reperfusion. Low body weight in HLBW and NLBW groups was accompanied by increased hematocrit, and decrease in absolute heart weight (both wet and dry) and developed force. On the other hand, higher hydration, increased cardiac tolerance to ischemia and potentiation of protection by ischemic preconditioning were observed in HLBW rats only. This experimental group also exhibited the highest relative heart weight. It may be concluded that low body weight alone does not influence cardiac tolerance to ischemia in neonatal rats.

Intermittent high altitude hypoxia protects the heart against lethal Ca 2+ overload injury

Adaptation to intermittent high altitude (IHA) hypoxia can protect the heart against ischemia-reperfusion injury. In view of the fact that both Ca 2+ paradox and ischemia-reperfusion injury are associated with the intracellular Ca 2+ overload, we tested the hypothesis that IHA hypoxia may protect hearts against Ca 2+ paradox-induced lethal injury if its cardioprotection bases on preventing the development of intracellular Ca 2+ overload. Langendorff-perfused hearts from normoxic and IHA hypoxic rats were subjected to Ca 2+ paradox (5 min of Ca 2+ depletion followed by 30 min of Ca 2+ repletion) and the functional, biochemical and pathological changes were investigated. The Ca 2+ paradox incapacitated the contractility of the normoxic hearts, whereas the IHA hypoxic hearts significantly preserved contractile activity. Furthermore, the normoxic hearts subjected to Ca 2+ paradox exhibited a marked reduction in coronary flow, increase in lactate dehydrogenase release, and severe myocyte damage. In contrast, these changes were significantly prevented in IHA hypoxic hearts. We, then, tested and confirmed our hypothesis that the protective mechanisms are mediated by mitochondria ATP-sensitive potassium channels (mitoK ATP) and Ca 2+/calmodulin-dependent protein kinase II (CaMKII), as the protective effect of IHA hypoxia was abolished by 5-hydroxydecanoate, a selective mitoK ATP blocker, and significantly attenuated by KN-93, a CaMKII inhibitor. In conclusion, our studies offer for the first time that IHA hypoxia confers cardioprotection against the lethal injury of Ca 2+ paradox and give biochemical evidence for the protective mechanism of IHA hypoxia. We propose that researches in this area may lead a preventive regimen against myocardial injury associated with Ca 2+ overload.

Phospholipid Composition of Immature Rat Myocardium Exposed to Chronic Hypoxia and the Effect of Normoxic Recovery

Four-day-old male Wistar rats were exposed to intermittent high-altitude (IHA) hypoxia of 7000 m simulated in a hypobaric chamber (8 h/day, 5 days/week, 25 exposures). The concentration of individual phospholipids (PL) and fatty acid (FA) composition of phosphatidylcholine (PC), phosphatidylethanolamine (PE) and diphosphatidylglycerol (DPG) were determined in right (RV) and left (LV) ventricles of rats adapted to chronic hypoxia (40-day-old), rats after 30 days of recovery from hypoxic to normoxic conditions (70-day-old) and both age-matched controls. The adaptation to IHA hypoxia decreased the concentration of DPG in LV (by 10%) in comparison with normoxic control. In hypoxic group the proportion of linoleic acid (18:2n-6) decreased; on the contrary, the proportion of arachidonic (20:4n-6), docosapentaenoic (22:5n-3) and docosahexaenoic (22:6n-3) acids increased in PC and PE of both RV and LV. As to DPG, IHA hypoxia caused a significant decrease in the n-6/n-3 ratio due to the increase in the 22:6n-3 proportion in RV. Thirty-day-long recovery from hypoxic to normoxic conditions led to complete regression of the hypoxic effect on FA composition in all PL. No difference in FA composition of PL was observed between RV and LV in any experimental group. Numerous dietary studies with fish oil supplements confirmed cardioprotective effect of n-3 polyunsaturated FA. We suppose that their increased content in heart-membrane PL observed in this study independently on a diet might contribute to higher tolerance of chronically hypoxic myocardium to ischemic injury.

Altered myocardial Gs protein and adenylyl cyclase signaling in rats exposed to chronic hypoxia and normoxic recovery.

The present work has analyzed the consequences of chronic intermittent high-altitude hypoxia for functioning of the G protein-mediated adenylyl cyclase (AC) signaling system in the right (RV) and left ventricular (LV) myocardium in rats. Adaptation to hypoxia did not appreciably affect the number of beta-adrenoceptors and the content of predominantly membrane-bound alpha-subunit (G(s)alpha) of the stimulatory G protein, but it raised the amount of cytosolic G(s)alpha in RV. The levels of myocardial inhibitory Galpha protein were not altered. Activity of AC stimulated by GTP, fluoride, forskolin, or isoprotertenol was reduced by approximately 50% in RV from chronically hypoxic rats, and a weaker depression was also found in LV. In addition, hypoxia significantly diminished a functional activity of membrane-bound G(s)alpha in both RV and LV. The RV baseline contractile function was markedly increased in chronically hypoxic animals, and its sensitivity to beta-adrenergic stimulation was decreased. Animals recovering from hypoxia for 5 wk still exhibited markedly elevated levels of cytosolic G(s)alpha and significantly lower activity of AC in RV than did age-matched controls, but contractile responsiveness to beta-agonists was normal.

Exposure to Intermittent High Altitude Induces Different Changes in Adenylyl Cyclase Activity in Hearts of Young and Adult Wistar Rats

This study investigates changes of adenylyl cyclase activity in the heart of young and adult Wistar rats exposed to experimental conditions simulating high altitude hypoxia as a model for interpretation of some adaptive changes of adenylyl cyclase observed in human. The exposure of rats to intermittent high altitude (IHA) hypoxia (5000 m) showed significant adaptive changes. The right ventricular weight and the ratio of right/left ventricular weights of adult rats exposed to IHA were significantly increased when compared to appropriate controls; adaptive changes of cardiac adenylyl cyclase being dependent on the age of the animals. The isoprenaline‐stimulated activity was higher in the left than in the right ventricle, and in both ventricles it was higher in young rats than in adult rats. When compared to controls, isoprenaline stimulation was decreased in the right ventricles of adapted young rats and, by contrast, it was increased in the left ventricles of adapted adult rats. This decrease and increase of adenylyl cyclase activity evoked by isoprenaline was paralleled by forskolin‐induced adenylyl cyclase activity in these experimental groups. It seems therefore that the changes in the pattern of total adenylyl cyclase activity observed under IHA hypoxia may at least be partially explained by the changes of beta‐adrenergic receptor susceptibility following IHA hypoxia.

Effect of intermittent high altitude hypoxia on gene expression in rat heart and lung

Hypoxia has been identified as an important stimulus for gene expression during embryogenesis and in various pathological situations. Its influence under physiological conditions, however, has only been studied occasionally. We therefore investigated the effect of intermittent high altitude hypoxia on the mRNA expression of different cytokines and protooncogenes, but also of other genes described to be regulated by hypoxia, in the left ventricle (LV), the right ventricle (RV), atria and the lung of adult rats after simulation of hypoxia in a barochamber (5000 m, 4 hours to 10 days). Heme oxygenase-1 as well as transforming growth factor-β1 showed an increased expression in all regions of the heart and the lung at different periods of hypoxia. For lactate dehydrogenase-A, we found a significant up-regulation in the RV and the lung, for lactate dehydrogenase-B up-regulation in the RV, but down-regulation in the LV and the atria. Vascular endothelial growth factor was up-regulated in the RV, the LV and the lung, but down-regulated in the atria. Its receptor Flk-1 mRNA was significantly increased in the atria and RV only. Expression of c-fos was found in the LV and RV only after 4 hours of hypoxia. The level of c-jun was significantly increased in the LV but decreased in the atria. Our data clearly demonstrate that intermittent hypoxia is a modulator of gene expression under physiological conditions. It differently regulates the expression of distinct genes not only in individual organs but even within one organ, i.e. in the heart.

Ischemic preconditioning in chronically hypoxic neonatal rat heart.

Rat hearts isolated on d 1, 4, 7, and 10 of postnatal life were perfused (in Langendorff mode) with Krebs-Henseleit solution at constant pressure, temperature, and stimulation rate. Recovery of the contractile function after global ischemia was measured by an isometric force transducer and analyzed using an online computer. Ischemic preconditioning (IP) was induced by three 3-min periods of global ischemia, each separated by a 5-min period of reperfusion. Prenatal hypoxia was induced by exposure of pregnant mothers to intermittent high altitude (IHA), simulated in a barochamber (8 h/d, 5000 m) from d 15 to 20 of pregnancy. Postnatal hypoxia was simulated by the identical procedure from postnatal d 1 to 6 and 9. Prenatal exposure to IHA failed to improve ischemic tolerance on d 1, but postnatal exposure to IHA improved recovery of the developed force after ischemia on d 7 (33 +/- 3% versus 43 +/- 4%) and 10 (39 +/- 2% versus 54 +/- 2%). Combination of IHA and IP induced higher protective effects in all age groups, including postnatal d 1 (48 +/- 2% versus 56 +/- 3%), whereas IHA and IP applied separately failed to improve ischemic tolerance. Neither the mitochondrial K(ATP) channel blocker 5-hydroxydecanoate nor the nitric oxide synthase inhibitor Nomega-nitro-L-arginine methyl ester abolished protection by IP in normoxic animals, but they decreased significantly protection by IHA hypoxia. The final recovery was even lower than the corresponding normoxic values. It seems likely that mitochondrial K(ATP) channels and nitric oxide may be involved in the protective mechanisms of adaptation to chronic hypoxia but not to that of IP, at least in neonates.

Effects of mitochondrial K(ATP) modulators on cardioprotection induced by chronic high altitude hypoxia in rats.

Adaptation of rats to intermittent high altitude hypoxia increases the tolerance of their hearts to acute ischemia/reperfusion injury. Our aim was to examine the role of mitochondrial ATP-sensitive potassium channels (K(ATP)) in this form of protection. Adult male Wistar rats were exposed to hypoxia of 5000 m in a barochamber for 8 h/day, 5 days a week; the total number of exposures was 24-32. A control group was kept under normoxic conditions (200 m). Infarct size (tetrazolium staining) was measured in anesthetized open-chest animals subjected to 20-min regional ischemia (coronary artery occlusion) and 4-h reperfusion. Isolated perfused hearts were used to assess the recovery of contractile function following 20-min global ischemia and 40-min reperfusion. In the open-chest study, a selective mitochondrial K(ATP) blocker, 5-hydroxydecanoate (5 mg/kg), or openers, diazoxide (10 mg/kg) or BMS-191095 (10 mg/kg), were administered into the jugular vein 5 and 10 min before occlusion, respectively. In the isolated heart study, 5-hydroxydecanoate (250 micromol/l) or diazoxide (50 micromol/l) were added to the perfusion medium 5 or 10 min before ischemia, respectively. In the control normoxic group, infarct size occupied 62.2+/-2.0% of the area at risk as compared with 52.7+/-2.5% in the chronically hypoxic group (P<0.05). Post-ischemic recovery of contractile function (dP/dt) reached 60.0+/-3.9% of the pre-ischemic value and it was improved to 72.4+/-1.2% by adaptation to hypoxia (P<0.05). While 5-hydroxydecanoate completely abolished these protective effects of chronic hypoxia, it had no appreciable influence in normoxic groups. In contrast, diazoxide significantly increased the recovery of contractile function and reduced infarct size in normoxic groups only. The later effect was also observed following treatment with BMS-191095. The results suggest that opening of mitochondrial K(ATP) channels is involved in the cardioprotective mechanism conferred by long-term adaptation to intermittent high altitude hypoxia.

Cardioprotective effects of chronic hypoxia and ischaemic preconditioning are not additive.

The objective of the work was to examine whether adaptation to intermittent high altitude hypoxia and ischaemic preconditioning provide additive protection of the heart against subsequent acute ischaemic injury. Adult male rats were exposed to hypoxia (7000 m, 8 h/day, 24-30 exposures) in a hypobaric chamber. Susceptibility of their hearts to ischaemia-induced ventricular arrhythmias and infarction was evaluated in open-chest animals subjected to 30-min coronary artery occlusion and 4-h reperfusion. Preconditioning was induced by either two (PC1) or five (PC2) occlusions of the same artery for 5 min, each followed by 5-min reperfusion. Adaptation to hypoxia decreased the arrhythmia score from 2.75 +/- 0.13 in normoxic controls to 2.17 +/- 0.18. Both PC1 and PC2 reduced the arrhythmia score in the controls (0.15 +/- 0.10 and 0.71 +/- 0.24, respectively), as well as in the hypoxic groups (0.40 +/- 0.15 and 0.27 +/- 0.15, respectively). The infarct size was reduced from 66.6 +/- 2.3% of the area at risk in the controls to 50.2 +/- 1.9% in the adapted rats. PC1 conferred further protection in adapted animals (38.4 +/- 2.8%) but this combined effect was of the same magnitude as that of preconditioning in the controls (37.5 +/- 1.6%). Similar results were obtained using PC2. It is concluded that adaptation to hypoxia decreases the efficiency of ischaemic preconditioning; cardioprotective effects of these two phenomena are not additive. The results are consistent with the view that the mechanisms of protection conferred by chronic hypoxia and preconditioning may share the same signalling pathway.

Chronic hypoxia alters fatty acid composition of phospholipids in right and left ventricular myocardium.

Adult male Wistar rats were exposed to intermittent high altitude hypoxia of 7000 m simulated in a hypobaric chamber for 8 h/day, 5 days a week; the total number of exposures was 25. The concentration of individual phospholipids and their fatty acid (FA) profile was determined in right (RV) and left (LV) ventricles. Adaptation to hypoxia decreased the concentration of diphosphatidytglycerol (DPG) in hypertrophied RV by 19% and in non-hypertrophied LV by 12% in comparison with normoxic controls. Chronically hypoxic hearts exhibited lower phospholipid n-6 polyunsaturated FA(PUFA) content mainly due to decreased linoleic acid (18:2n-6), which was opposed by increased n-3 PUFA mainly due to docosahexaenoic acid (22:6n-3) in phosphatidylcholine (PC), phosphatidylethanolamine (PE) and phosphatidylinositol (PI). The content of arachidonic acid (20:4n-6) was unchanged in total phospholipids, but in PC it was increased in both ventricles (by 22%) and in PE decreased in LV only (by 20%). Chronic hypoxia increased the un-saturation index of PC and PE in both ventricles. The content of monounsaturated FA (MUFA) was increased and 18:2n-6 decreased in DPG. The proportion of saturated FA was increased in PC and PI of hypoxic RV but not LV. The FA composition of phosphatidylserine was not altered in hypoxic ventricles. It is concluded that chronic hypoxia led to only minor changes in individual phospholipid concentration in rat ventricular myocardium, but markedly altered their FA profile. These changes, in particular the greater incorporation of n-3 PUFA into phospholipids and increased un-saturation index, may lead to a better preservation of membrane integrity and thereby contribute to improved ischemic tolerance of chronically hypoxic hearts.

Adaptation to High Altitude Hypoxia Protects the Rat Heart Against Ischemia-induced Arrhythmias. Involvement of Mitochondrial KATPChannel

The aim was to determine whether adaptation to chronic hypoxia protects the heart against ischemic arrhythmias and whether ATP-dependent potassium channels (KATP) play a role in the antiarrhythmic mechanism. Adult male rats were adapted to intermittent high altitude hypoxia (5000 m, 4 h/day) and susceptibility to ischemia-induced ventricular arrhythmias was evaluated in the Langendorff-perfused hearts subjected to either an occlusion of the coronary artery for 30 min or pre-conditioning by brief occlusion of the same artery prior to 30-min reocclusion. In separate groups, either a KATPblocker, glibenclamide (10 μ mol/l), or a mitochondrial KATPopener, diazoxide (50μ mol/l), were added to a perfusion medium 20 min before the occlusion. Adaptation to hypoxia reduced the total number of ventricular arrhythmias by 64% as compared with normoxic controls. Preconditioning by a single 3-min coronary artery occlusion was antiarrhythmic only in the normoxic group, while two occlusion periods of 5 min each were needed to pre-condition the hypoxic hearts. Glibenclamide increased the number of arrhythmias in the normoxic hearts from 1316±215 to 2091±187 (by 59%) and in the hypoxic group from 636±103 to 1777±186 (by 179%). In contrast, diazoxide decreased the number of arrhythmias only in the normoxic group from 1374±96 to 582±149 (by 58%), while its effect in the hypoxic group was not significant. It is concluded that long-term adaptation of rats to high altitude hypoxia decreases the susceptibility of their hearts to ischemic arrhythmias and increases an antiarrhythmic threshold of pre-conditioning. The mitochondrial KATPchannel, rather than the sarcolemmal KATPchannel, apears to be involved in the protective mechanism afforded by adaptation.

Activity of cytochrome c oxidase in the right and left ventricular myocardium of male and female rats exposed to intermittent high altitude hypoxia.

The aim of the present study was to compare the capacity of the oxidative metabolism (total activity of cytochrome c oxidase, COX) in the right and left ventricular myocardium of adult rats exposed to intermittent high altitude (IHA) hypoxia simulated in a barochamber (5,000 m, 8 h/day, 5 days/wk, for a total of 32 exposures). In male and female rats, IHA induced significant increases of the right ventricular (RV) weight and protein content, whereas left ventricular (LV) weight and protein content remained unaffected. Consequently, the RV/LV ratio in both sexes markedly increased. Similarly, IHA induced an increase of the total activity of COX in RV in both sexes. The specific activities of COX in homogenate as well as in isolated mitochondria were not changed in IHA-exposed animals, which indicates that the increase of total activity of COX is proportional to the increase of total protein content and RV weight.

Regression of chronic hypoxia-induced pulmonary hypertension, right ventricular hypertrophy, and fibrosis: effect of enalapril.

Chronic hypoxia induces pulmonary hypertension and right ventricular hypertrophy. These changes are completely reversible, except for persistent myocardial fibrosis. The aim of the present study was to determine whether treatment with the angiotensin-converting enzyme (ACE) inhibitor enalapril can reduce the ventricular collagen content in animals recovering from chronic hypoxia. Adult male Wistar rats were exposed to intermittent high-altitude hypoxia simulated in a barochamber (7000 m, 8 hr/day, 5 days a week, 24 exposures), then transferred to normoxia and divided into two groups: (a) treated with enalapril (0.1 g/kg/day for 60 days) and (b) without treatment. The corresponding control groups were kept under normoxic conditions. Enalapril significantly decreased the heart rate, systemic arterial pressure, and absolute left and right ventricular weights in both hypoxic and control rats; on the other hand, the pulmonary blood pressure was unchanged. The content and concentration of collagen was reduced in both ventricles of enalapril-treated hypoxic and control animals by 10-26% compared with the corresponding untreated groups. These data suggest that the partial regression of cardiac fibrosis due to enalapril may be independent of the pressure load.

Myoglobin as an indicator of stress damage

The effects of single 1-hour immobilization stress exposure and of long-term (40 days) adaptation to intermittent high altitude hypoxia (in hypobaric chamber at "altitude" 4000 m, 4 hr, a day) on serum myoglobin content were studied in Wistar rats. It has been shown that such stress exposure induced the 2.5 fold increase in the serum myoglobin content. The adaptation did not change the myoglobin content, but effectively limited mentioned above stress-induced increase in the serum myoglobin level. These data suggest that serum myoglobin content can be used as a criterion for stress damage and efficiency of factors protecting against stress damage.