Weeks Of Hypoxic Exposure Research Articles

Different cardiovascular and pulmonary phenotypes for single- and double-knock-out mice deficient in BMP9 and BMP10.

Aims BMP9 and BMP10 mutations were recently identified in patients with pulmonary arterial hypertension, but their specific roles in the pathogenesis of the disease are still unclear. We aimed to study the roles of BMP9 and BMP10 in cardiovascular homeostasis and pulmonary hypertension using transgenic mouse models deficient in Bmp9 and/or Bmp10.Methods and resultsSingle- and double-knockout mice for Bmp9 (constitutive) and/or Bmp10 (tamoxifen inducible) were generated. Single-knock-out (KO) mice developed no obvious age-dependent phenotype when compared with their wild-type littermates. However, combined deficiency in Bmp9 and Bmp10 led to vascular defects resulting in a decrease in peripheral vascular resistance and blood pressure and the progressive development of high-output heart failure and pulmonary hemosiderosis. RNAseq analysis of the lungs of the double-KO mice revealed differential expression of genes involved in inflammation and vascular homeostasis. We next challenged these mice to chronic hypoxia. After 3 weeks of hypoxic exposure, Bmp10-cKO mice showed an enlarged heart. However, although genetic deletion of Bmp9 in the single- and double-KO mice attenuated the muscularization of pulmonary arterioles induced by chronic hypoxia, we observed no differences in Bmp10-cKO mice. Consistent with these results, endothelin-1 levels were significantly reduced in Bmp9 deficient mice but not Bmp10-cKO mice. Furthermore, the effects of BMP9 on vasoconstriction were inhibited by bosentan, an endothelin receptor antagonist, in a chick chorioallantoic membrane assay.ConclusionsOur data show redundant roles for BMP9 and BMP10 in cardiovascular homeostasis under normoxic conditions (only combined deletion of both Bmp9 and Bmp10 was associated with severe defects) but highlight specific roles under chronic hypoxic conditions. We obtained evidence that BMP9 contributes to chronic hypoxia-induced pulmonary vascular remodelling, whereas BMP10 plays a role in hypoxia-induced cardiac remodelling in mice.

Effects of Intermittent Normoxia on Chronic Hypoxic Pulmonary Hypertension and Right Ventricular Hypertrophy in Rats.

Liu, Chunlei, Xu Chen, Ge Guo, Xiang Xu, Xin Li, Qingxia Wei, Yanying Shen, Hanlu Li, Jianxiu Hao, Ya Ping Tian, and Kunlun He. Effects of intermittent normoxia on chronic hypoxic pulmonary hypertension and right ventricular hypertrophy in rats. High Alt Med Biol. 22: 184-192, 2021. Background: Individuals with chronically low arterial oxygen tension owing to high altitude develop elevated rates of pulmonary hypertension (PH) and right ventricular (RV) hypertrophy. However, the effects of the frequency and duration of normoxic exposure on PH and RV hypertrophy have not been adequately assessed; thus, we aimed to analyze the same. Materials and Methods: PH and RV hypertrophy were induced in 60 rats using a hypobaric chamber. Of these 60 rats, every 10 were exposed to normoxic conditions for 30 minutes once (1T/D), three times (3T/D), or five times daily (5T/D), or for one 150-minute recovery daily (1LT/D). Furthermore, 10 rats were housed in a normoxic environment, and another 10 were subjected to continuous hypoxia. After 4 weeks, hemodynamic measurements were recorded, and the hearts were harvested for pathomorphological observations. Results: Average pulmonary arterial pressures (PAP) of control rats and those exposed to hypobaric hypoxia were 14.1 and 32.3 mmHg, respectively. After 30 minutes of exposure to normoxia 3T/D, 5T/D, or 1LT/D, PAP values were reduced to 27.1, 27.9, or 26.8 mmHg, respectively. Four weeks of hypoxic exposure elevated the RV/heart weight (HW) ratios, while exposure to normoxia 3T/D, 5T/D, and 1LT/D significantly reduced RV/HW. In addition, exposure to normoxia 3T/D, 5T/D, 1LT/D reduced the percentage wall thickness of the pulmonary artery as well as the hypertrophy indices of atrial natriuretic peptide, brain natriuretic peptide, and myosin heavy chain 7 (MYH-7). Conclusions: Thirty-minute exposure to normoxic conditions of 3T/D, 5T/D, or 1LT/D effectively ameliorates PH and RV thickening.

Hypoxic Exposure Increases Energy Expenditure by Increasing Carbohydrate Oxidation in Mice

Aims. Hypoxic exposure improves glucose metabolism. We investigated to validate the hypothesis that carbohydrate (CHO) oxidation could increase in mice exposed to severe hypoxic conditions.Methods. Seven-week-old male ICR mice (n=16) were randomly divided into two groups: the control group (CON) was kept in normoxic condition (fraction of inspiredO2=21%) and the hypoxia group (HYP) was exposed to hypoxic condition (fraction of inspiredO2=12%, ≈altitude of 4,300 m). The CON group was pair-fed with the HYP group. After 3 weeks of hypoxic exposure, we measured respiratory metabolism (energy expenditure and substrate utilization) at normoxic conditions for 24 hours using an open-circuit calorimetry system. In addition, we investigated changes in carbohydrate mechanism-related protein expression, including hexokinase 2 (HK2), pyruvate dehydrogenase (PDH), pyruvate dehydrogenase kinase 4 (PDK4), and regulator of the genes involved in energy metabolism (peroxisome proliferator-activated receptor gamma coactivator 1-alpha, PGC1α) in soleus muscle.Results. Energy expenditure (EE) and CHO oxidation over 24 hours were higher in the HYP group by approximately 15% and 34% (p<0.001), respectively. Fat oxidation was approximately 29% lower in the HYP group than the CON group (p<0.01). Body weight gains were significantly lower in the HYP group than in the CON group (CON vs. HYP;1.9±0.9vs.−0.3±0.9;p<0.001). Hypoxic exposure for 3 weeks significantly reduced body fat by approximately 42% (p<0.001). PDH and PGC1αprotein levels were significantly higher in the HYP group (p<0.05). Additionally, HK2 was approximately 21% higher in the HYP group.Conclusions. Hypoxic exposure might significantly enhance CHO oxidation by increasing the expression of PDH and HK2. This investigation can be useful for patients with impaired glucose metabolism, such as those with type 2 diabetes.

Assessing the effect of hypoxia on cardiac metabolism using hyperpolarized 13 C magnetic resonance spectroscopy.

Hypoxia plays a role in many diseases and can have a wide range of effects on cardiac metabolism depending on the extent of the hypoxic insult. Noninvasive imaging methods could shed valuable light on the metabolic effects of hypoxia on the heart in vivo. Hyperpolarized carbon‐13 magnetic resonance spectroscopy (HP 13C MRS) in particular is an exciting technique for imaging metabolism that could provide such information.The aim of our work was, therefore, to establish whether hyperpolarized 13C MRS can be used to assess the in vivo heart's metabolism of pyruvate in response to systemic acute and chronic hypoxic exposure.Groups of healthy male Wistar rats were exposed to either acute (30 minutes), 1 week or 3 weeks of hypoxia. In vivo MRS of hyperpolarized [1‐13C] pyruvate was carried out along with assessments of physiological parameters and ejection fraction. Hematocrit was elevated after 1 week and 3 weeks of hypoxia.30 minutes of hypoxia resulted in a significant reduction in pyruvate dehydrogenase (PDH) flux, whereas 1 or 3 weeks of hypoxia resulted in a PDH flux that was not different to normoxic animals. Conversion of hyperpolarized [1‐13C] pyruvate into [1‐13C] lactate was elevated following acute hypoxia, suggestive of enhanced anaerobic glycolysis. Elevated HP pyruvate to lactate conversion was also seen at the one week timepoint, in concert with an increase in lactate dehydrogenase (LDH) expression. Following three weeks of hypoxic exposure, cardiac metabolism of pyruvate was comparable with that observed in normoxia.We have successfully visualized the effects of systemic hypoxia on cardiac metabolism of pyruvate using hyperpolarized 13C MRS, with differences observed following 30 minutes and 1 week of hypoxia. This demonstrates the potential of in vivo hyperpolarized 13C MRS data for assessing the cardiometabolic effects of hypoxia in disease.

Chronically Hypoxic Fetal Lambs Supported by an Extra-Uterine Device Exhibit Mitochondrial Dysfunction and Elevations of Hypoxia Inducible Factor 1-Alpha

Introduction: We have recently developed an extra-uterine environment for neonatal development (EXTEND) capable of supporting premature fetal lambs and have been able to replicate hypoxic in utero conditions by controlling fetal oxygen delivery. In this study, we investigated the fetal mitochondrial response to hypoxia. Methods: Eight premature fetal lambs were delivered via hysterotomy and transitioned to extra-uterine support for 3 weeks. The lambs were divided into 2 groups: normoxic fetuses which maintained physiologic oxygen delivery and hypoxic fetuses in which oxygen delivery was significantly reduced. Control fetuses were delivered via hysterotomy but not cannulated. Measurements of mitochondrial membrane potential (MMP) were performed in peripheral blood mononuclear cells. Results: There were no significant differences in MMP between normoxic EXTEND fetuses and controls. Hypoxic fetuses had significantly more depolarized mitochondria compared to normoxic fetuses overall, and these changes were specifically appreciated in weeks 1 and 2, but not by week 3. Hypoxic fetuses had significantly elevated levels of HIF-1α compared to normoxic fetuses in the first 2 weeks. Discussion: Normoxic fetal lambs supported by EXTEND demonstrate normal mitochondrial function as evidenced by equivalent membrane potentials compared to control fetuses. Hypoxic fetuses exhibit mitochondrial dysfunction, though they do show evidence of adaptation after 3 weeks of hypoxic exposure.

Combination of erythropoietin and sildenafil can effectively attenuate hypoxia-induced pulmonary hypertension in mice.

Pulmonary hypertension (PH) is an incurable disease that often leads to right ventricular hypertrophy and right heart failure. This study investigated single versus combined therapy with sildenafil and erythropoietin on hypoxia-induced pulmonary hypertension in mice. Mice were randomized into 5 groups and exposed to either hypoxia (10% oxygen) or normoxia for a total of 5 weeks. Hypoxic mice were treated with saline solution, erythropoietin (500 IU/kg 3 times weekly), sildenafil (10 mg/kg daily), or a combination of the two drugs for the last 2 weeks of hypoxic exposure. We measured right ventricular pressures using right heart catheterization, and the ventilatory response to hypoxia was recorded via whole-body plethysmography. Histological analyses were performed to elucidate changes in pulmonary morphology and appearance of right heart hypertrophy. Plasma levels of cardiotrophin-1 and atrial natriuretic peptide were quantified. Treatment with either erythropoietin or sildenafil alone lowered the hypoxia-induced increase of pulmonary pressure and reduced pulmonary edema formation, pulmonary vascular remodeling, and right ventricular hypertrophy. Notably, the combination of the two drugs had the most prominent effect. Changes in cardiotrophin-1 and atrial natriuretic protein levels confirmed these observations. The combination treatment with erythropoietin and sildenafil demonstrated an attenuation of the development of hypoxia-induced PH in mice that was superior to that observed for either drug when given alone.

N-acetylcysteine inhibits hypoxic pulmonary hypertension most effectively in the initial phase of chronic hypoxia

Exposure to chronic hypoxia results in hypoxic pulmonary hypertension (HPH). In rats HPH develops during the first two weeks of exposure to hypoxia, then it stabilizes and does not increase in severity. We hypothesize that free radical injury to pulmonary vascular wall is an important mechanism in the early days of the hypoxic exposure. Thus antioxidant treatment just before and at the beginning of hypoxia should be more effective in reducing HPH than antioxidant therapy of developed pulmonary hypertension. We studied adult male rats exposed for 4 weeks to isobaric hypoxia (F iO2 = 0.1) and treated with the antioxidant, N-acetylcysteine (NAC, 20 g/l in drinking water). NAC was given “early” (7 days before and the first 7 days of hypoxia) or “late” (last two weeks of hypoxic exposure). These experimental groups were compared with normoxic controls and untreated hypoxic rats (3-4 weeks hypoxia). All animals kept in hypoxia had significantly higher mean pulmonary arterial blood pressure (PAP) than normoxic animals. PAP was significantly lower in hypoxic animals with early (27.1 ± 0.9 mmHg) than late NAC treatment (30.5 ± 1.0 mmHg, P < 0.05; hypoxic without NAC 32.6 ± 1.2 mmHg, normoxic controls 14.9 ± 0.7 mmHg). Early but not late NAC treatment inhibited hypoxia-induced increase in right ventricle weight and muscularization of distal pulmonary arteries assessed by quantitative histology. We conclude that release of free oxygen radicals in early phases of exposure to hypoxia induces injury to pulmonary vessels that contributes to their structural remodeling and development of HPH.

Morphological changes in the rat carotid body 1, 2, 4, and 8 weeks after the termination of chronically hypocapnic hypoxia.

Morphological changes in the rat carotid bodies 1, 2, 4, and 8 weeks after the termination of chronically hypocapnic hypoxia (10% O2 for 8 weeks) were examined by means of morphometry and immunohistochemistry. The rat carotid bodies after 8 weeks of hypoxic exposure were enlarged several fold with vascular expansion. The carotid bodies 1 and 2 weeks after the termination of 8 weeks of hypoxic exposure were diminished in size, although their diameter remained larger than the normoxic controls. The expanded vasculature in chronically hypoxic carotid bodies returned to the normoxic control state. In the carotid bodies 1 week after the termination of chronic hypoxia, the density of NPY fibers was remarkably increased and that of VIP fibers was dramatically decreased in comparison with the density in chronically hypoxic carotid bodies. In the carotid bodies 2 and 4 weeks after the termination of hypoxia, the density of SP and CGRP fibers was gradually increased. In the carotid bodies 8 weeks after the termination of hypoxia, the appearance of the carotid body returned to a nearly normoxic state, and the density of SP, CGRP, VIP, and NPY fibers also recovered to that of normoxic controls. These results suggest that the morphological changes in the recovering carotid bodies start at a relatively early period after the termination of chronic hypoxia, and a part of these processes may be under the control of peptidergic innervation.

Peptidergic innervation in the rat carotid body after 2, 4, and 8 weeks of hypocapnic hypoxic exposure.

The distribution and abundance of neuropeptide-containing nerve fibers were examined in the carotid bodies of rats exposed to hypocapnic hypoxia (10% O2 in N2) for 2, 4, and 8 weeks. The carotid bodies after 2, 4, and 8 weeks of hypoxic exposure were enlarged by 1.2-1.5 times in the short axis, and 1.3-1.7 times in the long axis in comparison with the normoxic control ones. The enlarged carotid bodies contained a number of expanded blood vessels. Mean density per unit area (10(4) microm2) of substance P (SP) and calcitonin gene-related peptide (CGRP) immunoreactive fibers was transiently high in the carotid bodies after 4 weeks of hypoxic exposure, and decreased significantly to nearly or under 50% after 8 weeks of hypoxic exposure. Density of vasoactive intestinal polypeptide (VIP) immunoreactive fibers increased significantly in all periods of hypoxic exposure observed, and was especially high in the carotid bodies after 4 weeks of hypoxic exposure. Density of neuropeptide Y immunoreactive fibers was unchanged in the carotid bodies during hypoxic exposure. These characteristic changes in the density of SP, CGRP, and VIP fibers in the carotid bodies after 4 weeks of hypoxic exposure suggest that the role of these neuropeptide-containing fibers may be different in the carotid bodies after each of three periods of hypoxic exposure, and that the peptidergic innervation after 8 weeks of hypoxic exposure may show an acclimatizing state.

Role of HIF-1 in physiological adaptation of the carotid body during chronic hypoxia.

Chemoreceptors in the carotid body (CB) increase intracellular calcium and afferent nerve discharge in hypoxia and, thus, play an important role in the regulation of cardiorespiratory responses to hypoxia (see Gonzalez et al 1994 for review). Also, the CB plays a central role in the ventilatory acclimatization to hypoxia involving an initial rapid increase in ventilation followed by a progressive hyperventilation during hours and weeks of hypoxic exposure (see Bisgard, 2000; Lahiri et al 2001 for reviews). It is well known that the CB enlarges in human and animals living at high altitude or exposed to chronic hypoxia. The enlargement may be due to an increased vasculature, cellular hypertrophy and hyperplasia of the glomus cells in the CB (Dhillon et al. 1984; McGregor et al 1984; Bee et al 1986).

Two-week, but not 1-week, hypoxic exposure enhances nitric oxide-mediated basal tone regulation in rat resistance pulmonary arteries.

We measured internal diameter (ID) changes in resistance and conduit pulmonary arteries of 1- and 2-week hypoxic rats and normoxic control rats in response to nitric oxide synthase (NOS) inhibitors in vivo. At 2 weeks of hypoxic exposure, the ID reduction as a result of NOS inhibition was enhanced within the resistance arteries, but not at 1 week of hypoxia.

Decreased rat brain cytochrome oxidase activity after prolonged hypoxia

Three weeks of hypoxic exposure results in a spectrum of systemic physiological and local brain tissue adaptations. Cytochrome oxidase histochemistry was used as an indicator of changes in energy demand in response to hypoxia. We found overall cytochrome oxidase activity decreased in hypoxic adapted rats as compared to normoxic control rats. Some regions, notably layer 3 of the frontal cerebral cortex, layer 4 of the parietal sensory barrel fields, lacunosum moleculare hippocampi, and specific nuclei of the rostral ventral medulla, though exhibiting cytochrome oxidase activity decreases of 16–27%, still retain their relatively higher levels of activity. We conclude that there is a hypometabolic component of the rat brain adaptation to continued hypoxia.

Increased basic fibroblastic growth factor mRNA in the brains of rats exposed to hypobaric hypoxia.

Continued exposure of rats to hypobaric hypoxia results in both functional and structural changes in brain. A most striking structural change is the 50-80% increased capillary density in the cerebral cortex and other brain regions after 3 weeks of hypoxic exposure (LaManna et al., 1992; LaManna, 1992). The apparent increased density may be accompanied by a shift in the capillary segment length distribution towards longer capillary segments (LaManna et al., 1993). It is still not clear if the increased capillary density is due to new capillary branching or to elongation of existing capillaries, or both. Nevertheless, it seems likely that the microvascular adaptation to continued hypoxia is controlled by growth factors. The growth factors that control angiogenesis are not yet well understood, but basic fibroblast growth factor (bFGF) is known to activate endothelial cells resulting in capillary growth (de Juan et al., 1990), and this growth factor is increased following metabolic stress such as brain ischemia (Finkelstein et al., 1990; Lyons et al., 1991). In this study, we determined whether bFGF mRNA was increased in brains of rats exposed to 3 weeks of hypoxia.

長期低酸素曝露による肺高血圧招来における血小板活性化因子の関与

Chronic hypoxic exposure causes pulmonary hypertension and right ventricular hypertrophy associated with pulmonary vascular remodeling in rats. Since platelet-activating factor (PAF) production has been shown to increase during hypoxia, and since PAF contributes to pulmonary hypertension, we hypothesized that increased PAF during hypoxia might play a role in the development of pulmonary hypertension induced by hypoxic exposure, and examined the effect of PAF-antagonists on hypoxia-induced pulmonary hypertension. Pulmonary hypertension and right ventricular hypertrophy developed at 3 weeks of hypoxic exposure. Treatment with WEB 2170 or BN 50739, specific PAF-antagonists significantly reduced hypoxia-induced pulmonary hypertension. Hypoxia-induced pulmonary hypertension was associated with an increase in the vessel wall thickness of the muscular arteries. In WEB 2170-treated rats, this change was significantly less severe than that observed in untreated chronically hypoxic rats. These results indicate that the PAF-antagonists inhibit the development of pulmonary hypertension induced by chronic hypoxic exposure and suggest a role for PAF in the remodeling process of pulmonary vessels induced by chronic hypoxia in rats.

Hypotensive effect of heparin on experimental chronic pulmonary hypertension in rats.

Daily heparin injections (300 mg s.c.) during four weeks of hypoxic exposure resulted in a smaller rise in pulmonary artery pressure as compared with otherwise nontreated rats exposed to the same regimen of hypoxia. Heparin treatment did not significantly affect the systemic arterial pressure of the hypoxic rats. In spite of the different pressure load of the right ventricle after hypoxia in treated and control rats, there was no difference in parameters indicative of the degree of right ventricular hypertrophy between these two groups of hypoxic rats. This finding suggests that the difference in pressure load between the two groups of hypoxic rats occurred late in the course of hypoxic exposure.

Hematology and red cell morphology in guinea pigs acclimated to chronic hypoxia during growth

1. 1. Weanling male guinea pigs were exposed for up to 14 weeks to an ambient P o 2 of 80 torr. Hematological measurements made on these animals were compared to data for normoxic controls of similar age and body weight. 2. 2. Within 2 weeks of hypoxic exposure, hematocrit was increased by 42% and [Hb] by 41% compared to controls. However, these changes were accompanied by only a 13% increase in the No. RBC's/mm 3 during the same period; mean red cell volume and mean cell hemoglobin content were each increased by 22% compared to controls. Normochromic macrocytes as well as numerous nucleated RBC's were evident in the peripheral smears of the hypoxia acclimated animals. 3. 3. Frequency analysis of RBC diameters showed a single peak at 6.3 μm for all controls. In guinea pigs acclimated to P O 2 = 80 torr, RBC diameters were initially bimodally distributed, with peaks at 6.3 and 7.6 μm, and later were broadly distributed with a mean of 7.0 μm. 4. 4. These findings support the hypothesis that, under severe and chronic erythropoietic stress, normochromic macrocytes appear in the peripheral blood following a skipped terminal mitotic division, during which cytoplasmic volume would normally be reduced.