High Altitude Rats Research Articles

Effect of High Altitude Environment on Pharmacokinetic and Pharmacodynamic of Warfarin in Rats.

High altitude environment affects the pharmacokinetic (PK) parameters of drugs and the PK parameters are an important theoretical basis for guiding the rational clinical use of drugs. Warfarin is an oral anticoagulant of the coumarin class commonly used in clinical practice, but it has a narrow therapeutic window and wide individual variation. However, the effect of high altitude environment on PK and pharmacodynamic (PD) of warfarin is unclear. The objective of this study is to investigate the effect of a high altitude environment on PK and PD of warfarin in rats. Rats were randomly divided into plain group and high altitude group and blood samples were collected through the orbital venous plexus after administration of 2 mg/kg warfarin. Warfarin concentrations in plasma samples were determined by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and PK parameters were calculated by the non-compartment model using WinNonlin 8.1 software. Meanwhile, the expression of PXR, P-gp and CYP2C9 in liver tissues was also determined by western blotting. The effect of high altitude environment on PD of warfarin was explored by measuring activated partial thromboplastin time (APTT) and prothrombin time (PT) values and then calculated international normalized ratio (INR) values based on PT. Significant changes in PK behaviors and PD of warfarin in high altitude-rats were observed. Compared with the plain-rats, the peak concentration (Cmax) and the area under the plasma concentration-time curve (AUC) increased significantly by 50.9% and 107.46%, respectively. At the same time, high altitude environment significantly inhibited the expression of PXR, P-gp and CYP2C9 in liver tissues. The results of the PD study showed that high altitude environments significantly prolonged PT, APTT and INR values. High altitude environment inhibited the metabolism and increased the absorption of warfarin in rats and increased the effect of anticoagulant effect, suggesting that the optimal dose of warfarin for patients at high altitude should be reassessed.

Discussion on the relationship between the disposal time of hypobaric oxygen chamber and the establishment of rat cardiac arrest model at high altitude

To establish the rat cardiac arrest model in high-altitude hypobaric hypoxia environment, and to explore the effect of the treatment time in the hypobaric oxygen chamber on the reproduction of high-altitude rat cardiac arrest model. SPF grade healthy male Sprague-Dawley (SD) rats were used as observation subjects. The experiment was conducted in two different altitude areas. The rats from the Plateau Branch of Institute of Cardiopulmonary and Cerebral Resuscitation of Sun Yat-sen University (Xining, Qinghai) were weighed and numbered, and they were placed in a hypobaric oxygen chamber (simulated altitude of 3 000 meters, speed of ascent and descent of 15 m/min, temperature of 20 centigrade, cabin pressure of 69.5 kPa, cabin oxygen pressure of 14.5 kPa). After 30 days of feeding, the rats were obtained according to random number table method, and the cardiac arrest model was established by asphyxia method as the 30-day hypobaric hypoxia group. After 60 days of feeding, rats were randomly selected again, and the cardiac arrest model was established as the 60-day hypobaric hypoxia group. Thirty rats were randomly selected from the Institute of Cardiopulmonary Cerebral Resuscitation at Sun Yat-sen University (Guangzhou, Guangdong) by the same method, and the cardiac arrest model was established as the plain control group. The differences in the body weight of rat modeling precursors and the induction time of asphyxia during the modeling process among different groups were compared. Finally, cardiac arrest model was established in 16 rats in the 30-day hypobaric hypoxia group and in 22 rats in the 60-day hypobaric hypoxia group. There was no significant difference in the body weight of rats before modeling among the plain control group, 30-day hypobaric hypoxia group and 60-day hypobaric hypoxia group [g: 429.00 (389.25, 440.75), 440.00 (415.50, 486.25), 440.00 (400.00, 452.50), all P > 0.05]. The asphyxia induction time of rats in the 60-day hypobaric hypoxia group was significantly longer than that in the 30-day hypobaric hypoxia group (s: 294.59±75.39 vs. 234.31±93.86, P < 0.01), even about 1.4 times of the plain control group (s: 294.59±75.39 vs. 208.73±30.88, P < 0.01). There was no significant difference in the asphyxia induction time between the 30-day hypobaric hypoxia group and the plain control group (P > 0.05). Rats treated in a hypobaric oxygen chamber for 60 days are more suitable for the preparation of high-altitude cardiac arrest model, and are also consistent with the oxygen reserve and hypoxia tolerance of high-altitude rats.

Effects of high-altitude environment on pharmacokinetic parameters of gliquidone in rats.

To investigate the effect of high-altitude hypoxia on the pharmacokinetics parameters of gliquidone. Twelve healthy male Wistar rats were randomly divided into plain group and high-altitude group with 6 rats in each group. Blood samples were collected after intragastric administration of gliquidone (6.3 mg/kg). Ultra-fast liquid chromatography-tandem mass spectrometry (UFLC-MS/MS) was used to determine the concentration of gliquidone in rat plasma samples. And the expression of CYP2C9 in rat liver tissues was determined by Western blotting. Compared with the plain group, the peak concentration of gliquidone in the high-altitude rats was significantly increased, the absorption rate constant was decreased, the elimination rate constant and the absorption half-life were increased, the elimination half-life was shortened, the mean residence time and apparent volume of distribution were decreased (all P<0.05). Western blotting showed that the expression of CYP2C9 was significantly up-regulated in the liver tissues of high altitude group rats, compared with the plain group (4.18 ±0.06 vs. 2.13±0.06, t=11.57, P<0.01). Under the high-altitude hypoxia environment, the absorption of gliquidone in rats was reduced and the metabolism was accelerated in rats, which may be related to the up-regulation of CYP2C9 expression in liver tissues.

Divergent Mitochondrial Antioxidant Activities and Lung Alveolar Architecture in the Lungs of Rats and Mice at High Altitude.

Compared with mice, adult rats living at 3,600 m above sea level (SL—La Paz, Bolivia) have high hematocrit, signs of pulmonary hypertension, and low lung volume with reduced alveolar surface area. This phenotype is associated with chronic mountain sickness in humans living at high altitude (HA). We tested the hypothesis that this phenotype is associated with impaired gas exchange and oxidative stress in the lungs. We used rats and mice (3 months old) living at HA (La Paz) and SL (Quebec City, Canada) to measure arterial oxygen saturation under graded levels of hypoxia (by pulse oximetry), the alveolar surface area in lung slices and the activity of pro- (NADPH and xanthine oxidases—NOX and XO) and anti- (superoxide dismutase, and glutathione peroxidase—SOD and GPx) oxidant enzymes in cytosolic and mitochondrial lung protein extracts. HA rats have a lower arterial oxygen saturation and reduced alveolar surface area compared to HA mice and SL rats. Enzymatic activities (NOX, XO, SOD, and GPx) in the cytosol were similar between HA and SL animals, but SOD and GPx activities in the mitochondria were 2–3 times higher in HA vs. SL rats, and only marginally higher in HA mice vs. SL mice. Furthermore, the maximum activity of cytochrome oxidase-c (COX) measured in mitochondrial lung extracts was also 2 times higher in HA rats compared with SL rats, while there was only a small increase in HA mice vs. SL mice. Interestingly, compared with SL controls, alterations in lung morphology are not observed for young rats at HA (15 days after birth), and enzymatic activities are only slightly altered. These results suggest that rats living at HA have a gradual reduction of their alveolar surface area beyond the postnatal period. We can speculate that the elevation of SOD, GPx, and COX activities in the lung mitochondria are not sufficient to compensate for oxidative stress, leading to damage of the lung tissue in rats.

Vitamins E and C co-administration attenuates acute swimming-induced hepatorenal oxidative damage and improves lipid profile in untrained Wistar rats native to low and high altitude

The aim of this study was to evaluate the effect of co- administration of vitamins E and C on exhaustive exercise induced-stress in regards to hepatorenal function in rats native to low altitude (LA) and high altitude (HA). In both LA and HA areas, native wistar rats of each area were divided into three groups of 6 rats each, which include stress-free control, forced swimming-induced experimental stress and experimental stress plus vitamins E and C treatment. Lipid profile and Liver and kidney functions were assessed in both groups. HA and LA rats exhibit similar baseline levels of liver and kidney function as well as lipid metabolism profiles. However, HA rats showed decreased levels of antioxidant markers with an increased level of lipid peroxidation. Exhaustive swimming exercise induced a significant increase in the liver and kidney function of rats at both altitudes accompanied with a decrease in antioxidants levels. However, the magnitude of change observed in HA rats was more profound. Also at LA, forced swimming exercise resulted in a significant increase in serum total cholesterol (TChol), triacylglycerides (TAG) and high-density lipoprotein cholesterol (HDL). However, in HA rats, forced swimming exercise caused a significant decrease in serum TChol and low-density lipoprotein (LDL), except for HDL levels which were significantly elevated. Pre- and co-administration of vitamins E and C counteracted the induction of liver and/or kidney function by exhaustive exercise, and lowered TChol and LDL levels in rats at either altitude. In conclusion, at native high altitude: kidney and liver function essentially remained stable; response to stress included more profound oxidative damage to liver and kidney tissues as well as augmented deterioration in lipid metabolism compared to low altitude; and combined administration of vitamins E and C protected against observed oxidative stress damage to liver and kidney tissues and preserved lipid metabolism. At low altitude, combined administration of vitamin E and C protected against stress-induced oxidative damage to the liver and kidney and did preserve normal lipid metabolism, except for HDL. These novel findings reveal the pathophysiological changes in the liver function, kidney function and lipid metabolism occurring at high altitude specifically under stress, and demonstrate the efficacy of combined supplementation of vitamins E and C to normalize these changes. Key words: Exercise, oxidative stress, vitamin E, vitamin C, altitude, rats.

Co-administration of Vitamins E and C Protects Against Stress-induced Hepatorenal Oxidative Damage and Effectively Improves Lipid Profile at both Low and High Altitude

The aim of this study was to evaluate the effect of co- administration of vitamins E and C on exhaustive exercise induced-stress in regards to hepatorenal function in rats native to low altitude (LA) and high altitude (HA). In both LA and HA areas, native wistar rats of each area were divided into three groups of 6 rats each, which include stress-free control, forced swimming-induced experimental stress and experimental stress plus vitamins E and C treatment. Lipid profile and Liver and kidney functions were assessed in both groups. HA and LA rats exhibit similar baseline levels of liver and kidney function as well as lipid metabolism profiles. However, HA rats showed decreased levels of antioxidant markers with an increased level of lipid peroxidation. Exhaustive swimming exercise induced a significant increase in the liver and kidney function of rats at both altitudes accompanied with a decrease in antioxidants levels. However, the magnitude of change observed in HA rats was more profound. Also at LA, forced swimming exercise resulted in a significant increase in serum total cholesterol (TChol), triacylglycerides (TAG) and high-density lipoprotein cholesterol (HDL). However, in HA rats, forced swimming exercise caused a significant decrease in serum TChol and low-density lipoprotein (LDL), except for HDL levels which were significantly elevated. Pre- and co-administration of vitamins E and C counteracted the induction of liver and/or kidney function by exhaustive exercise, and lowered TChol and LDL levels in rats at either altitude. In conclusion, at native high altitude: kidney and liver function essentially remained stable; response to stress included more profound oxidative damage to liver and kidney tissues as well as augmented deterioration in lipid metabolism compared to low altitude; and combined administration of vitamins E and C protected against observed oxidative stress damage to liver and kidney tissues and preserved lipid metabolism. At low altitude, combined administration of vitamin E and C protected against stress-induced oxidative damage to the liver and kidney and did preserve normal lipid metabolism, except for HDL. These novel findings reveal the pathophysiological changes in the liver function, kidney function and lipid metabolism occurring at high altitude specifically under stress, and demonstrate the efficacy of combined supplementation of vitamins E and C to normalize these changes. Key words: Exercise, oxidative stress, vitamin E, vitamin C, altitude, rats.

Life-long consequences of postnatal normoxia exposure in rats raised at high altitude

We tested the hypothesis that exposure of high-altitude (HA) rats to a period of postnatal normoxia has long-term consequences on the ventilatory and hematological acclimatization in adults. Male and female HA rats (3,600 m, Po(2) ≃ 100 Torr; La Paz, Bolivia) were exposed to normal room air [HA control (HACont)] or enriched oxygen (32% O(2); Po(2) ≃ 160 Torr) from 1 day before to 15 days after birth [HA postnatal normoxia (HApNorm)]. Hematocrit and hemoglobin values were assessed at 2, 12, and 32 wk of age. Cardiac and lung morphology were assessed at 12 wk by measuring right ventricular hypertrophy (pulmonary hypertension index) and lung air space-to-tissue ratio (indicative of alveolarization). Respiratory parameters under baseline conditions and in response to 32% O(2) for 10 min (relieving the ambient hypoxic stimulus) were measured by whole body plethysmography at 12 wk. Finally, we performed a survival analysis up to 600 days of age. Compared with HACont, HApNorm rats had reduced hematocrit and hemoglobin levels at all ages (both sexes); reduced right ventricular hypertrophy (both sexes); lower air space-to-tissue ratio in the lungs (males only); reduced CO(2) production rate, but higher oxygen uptake (males only); and similar respiratory frequency, tidal volume, and minute ventilation. When breathing 32% O(2), HApNorm male rats had a stronger decrease of minute ventilation than HACont. HApNorm rats had a marked tendency toward longer survival throughout the study. We conclude that exposure to ambient hypoxia during postnatal development in HA rats has deleterious consequences on acclimatization to hypoxia as adults.

Enhanced erythropoietin expression in the brainstem of newborn rats at high altitude

In addition to its role in elevating red blood cell number, erythropoietin (Epo) exerts protective functions against acute and delayed degenerative diseases of the brain. Moreover, we have recently demonstrated that endogenously synthesized Epo and soluble Epo receptor (a negative regulator of Epo binding to the Epo receptor) in the central nervous system play a crucial role in facilitating the ventilatory response and acclimatization to hypoxia. Here we hypothesized that cerebral Epo in the brainstem is implicated in the process that allows cardiorespiratory acclimatization to high altitude hypoxia during the postnatal period. Thus, we evaluated the postnatal ontogeny of cerebral Epo concentration of Sprague-Dawley rats living and reproducing at high altitude for longer than 19 years (3600 m in La Paz, Bolivia). Our results show that postnatal Epo concentration in high-altitude rats is higher in the brainstem than in the forebrain. Moreover, although Epo concentration in the forebrain of high-altitude rats is similar to sea-level controls, Epo level in the brainstem is surprisingly 2-fold higher in high-altitude rats than in sea-level controls. These findings strongly suggest that brainstem Epo plays an important role in tolerance to high altitude hypoxia after birth. From a clinical perspective, a better understanding of the role of Epo in the postnatal development of cardiorespiratory responses in neonates exposed to acute or chronic hypoxia might be useful.

Dichloroacetate Inhibits Vascular Remodeling and Increases Voltage-gate K+ Expression in a High-altitude-induced Pulmonary Artery Hypertension Rat Model

To investigate the effect of dichloroacetate (DCA) on the mean pulmonary artery pressure (mPAP), pulmonary artery (PA) remodeling and voltage-gate K+ (Kv) channel expression in pulmonary arterial smooth muscle cells (PASMCs) in high altitude-induced pulmonary artery hypertension (HA-PAH) rats. Sprague-Dawley rats were randomly assigned to normal control (N), high altitude (HA), and HA+DCA (70 mg/kg DCA administration daily) groups (n=8 each). Rats were housed in a hypobaric, hypoxic chamber to mimic an altitude of 5000m for 21 days; then the mPAP and the wall thickness (WT) of the PA smooth muscle were measured. PASMCs apoptosis was examined using terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) stain. Real-time PCR, immunohistochemistry and western blot analyses were carried out to detect Kv1.5 and Kv2.1 expression in PASMCs. The expression of Kv1.5 and Kv2.1 was decreased in HA rats. With DCA treatment, the expression of Kv1.5 and Kv2.1 was restored, and the established HA-PAH was ameliorated. Compared with the HA-PAH rats, the DCA-treated rats displayed a decreased mPAP, WT of the PAs, right ventricular hypertrophy and ([Ca2+]i), and more PASMCs were apoptotic. DCA partially reversed the downregulation of Kv1.5 and Kv2.1 in the PASMCs of HA-PAH rats. DCA can reverse the remodeling of the PA and upregulate Kv1.5 and Kv2.1 expression in the PASMCs of HA-PAH rats. This result suggests that DCA may be an effective drug for treating HA-PAH and that restoring Kv1.5 and Kv2.1 can partially decrease mPAP.

Long-term ventilatory adaptation and ventilatory response to hypoxia in plateau pika (Ochotona curzoniae): role of nNOS and dopamine

We assessed ventilatory patterns and ventilatory responses to hypoxia (HVR) in high-altitude (HA) plateau pikas, repetitively exposed to hypoxic burrows, and control rats. We evaluated the role of neuronal nitric oxide synthase (nNOS) and dopamine by using S-methyl-l-thiocitrulline (SMTC) inhibitor and haloperidol antagonist, respectively. Ventilation (Vi) was measured using a whole body plethysmograph in conscious pikas (n = 9) and low-altitude (LA) rats (n = 7) at different Pi(O(2)) (56, 80, 111, 150, and 186 mmHg) and in HA acclimatized rats (n = 9, 8 days at 4,600 m) at two different Pi(O(2)) (56 and 80 mmHg). The effects of NaCl, SMTC, and haloperidol on ventilatory patterns were assessed in pikas at Pi(O(2)) = 56 and 80 mmHg. We observed a main species effect with larger Vi, tidal volume (VT), inspiratory time/total time (T(i)/T(tot)), and a lower expiratory time in pikas than in LA rats. Pikas had also a larger VT and lower respiratory frequency compared with HA rats in hypoxia. HVR of pikas and rats were not statistically different. In pikas, SMTC induced a significant increase in Vi and VT for a Pi(O(2)) of 56 mmHg, but had no effect for a PiO(2) of 80 mmHg, i.e., the living altitude of pikas. In pikas, haloperidol injection had no effect on any ventilatory parameter. Long-term ventilatory adaptation in pikas is mainly due to an improvement in respiratory pattern (VT and T(i)/T(tot)) with no significant improvement in HVR. The sensitivity to severe acute hypoxia in pikas seems to be regulated by a peripheral nNOS mechanism.

Dehydroepiandrosterone upregulates soluble guanylate cyclase and inhibits hypoxic pulmonary hypertension

It has been reported that dehydroepiandrosterone is a pulmonary vasodilator and inhibits chronic hypoxia-induced pulmonary hypertension. Additionally, dehydroepiandrosterone has been shown to improve systemic vascular endothelial function. Thus, we hypothesized that chronic treatment with dehydroepiandrosterone would attenuate hypoxic pulmonary hypertension by enhancing pulmonary artery endothelial function. Rats were randomly assigned to five groups. Three groups received food containing 0, 0.3, or 1% dehydroepiandrosterone during a 3-wk-exposure to simulated high altitude (HA). The other 2 groups were kept at Denver's low altitude (LA) and received food containing 0 or 1% dehydroepiandrosterone. Dehydroepiandrosterone dose-dependently inhibited hypoxic pulmonary hypertension (mean pulmonary artery pressures after treatment with 0, 0.3, and 1% dehydroepiandrosterone=45+/-5, 33+/-2*, and 25+/-1*# mmHg, respectively. *P<0.05 vs. 0% and # vs. 0.3%). Dehydroepiandrosterone (1%, 3 wks) treatment started after rats had been exposed to 3-wk hypoxia also effectively reversed established hypoxic pulmonary hypertension. Pulmonary artery rings isolated from both LA and HA rats treated with 1% dehydroepiandrosterone showed enhanced relaxations to acetylcholine and sodium nitroprusside, but not to 8-bromo-cGMP. In the pulmonary artery tissue from dehydroepiandrosterone-treated LA and HA rats, soluble guanylate cyclase, but not endothelial nitric oxide synthase, protein levels were increased. These results indicate that the protective effect of dehydroepiandrosterone against hypoxic pulmonary hypertension may involve upregulation of pulmonary artery soluble guanylate cyclase protein expression and augmented pulmonary artery vasodilator responsiveness to nitric oxide.

Carbohydrate utilization during exercise after high-altitude acclimation: a new perspective.

At high altitude (HA), carbohydrate (CHO) is thought to be the preferred fuel because of its higher yield of ATP per mole of O2. We used indirect calorimetry and D-[6-3H]glucose infusions to determine total CHO and circulatory glucose utilization during exercise in HA-acclimated and sea level (SL) rats. We hypothesized that the percent contribution of CHO to total metabolism (VO2) is determined by exercise intensity relative to an aerobic maximum (% VO2max). HA rats run under hypoxia (FIO2 = 0.12) showed a decrease in VO2max compared with SL (67.55 +/- 1.26 vs. 89.30 +/- 1.23 ml kg-1 min-1). When exercised at 60% of their respective VO2max, both groups showed the same relative use of CHO (38 +/- 3% and 38 +/- 5% of VO2, at the beginning of exercise, in HA and SL, respectively). In both HA and SL, circulatory glucose accounted for approximately 20% of VO2, the balance was provided by muscle glycogen (approximately 18% of VO2). After 20 min at a higher intensity of 80% VO2max, 54 +/- 5% (HA) and 59 +/- 4% (SL) of VO2 was accounted for by CHO. We conclude the following: (i) the relative contributions of total CHO, circulatory glucose, and muscle glycogen do not increase after HA acclimation because the O2-saving advantage of CHO is outweighed by limited CHO stores; and (ii) relative exercise intensity is the major determinant of metabolic fuel selection at HA, as well as at SL.

The effect of high altitude and caloric restriction on reactive carbonyl derivatives and activity of glutamine synthetase in rat brain

Both exposure to high altitude (HA) and caloric restriction (CR) may influence free radical generation. The aim of the present study was to evaluate the effects of 4 wk chronic exposure to 4,000m of HA and CR (60% of CR of control (C) rats) on protein oxidation in brain. Eighteen rats with similar body mass were assigned to CR, HA and C rats. Reactive carbonyl derivatives (RCD), marker of protein oxidation, were measured by Western blot. In addition, the activity and protein content of glutamine syntethase (GS) were determined. The body mass of C rats was significantly higher (P < 0.001) than that of HA and CR groups. The quantified signal intensity of RCD was significantly stronger in C rats than in HA and CR rats. The activity of GS was significantly increased in CR rats, while the protein content of GS was decreased in HA rats compared to C group. The data suggest mat both HA and CR decreases the accumulation of RCD in the brain, however the mechanism of the decrease seems to be different during HA and CR.

Role of sex hormones in development of chronic mountain sickness in rats.

After chronic exposure to hypoxia, Hilltop Sprague-Dawley rats developed excessive polycythemia and severe pulmonary hypertension and right ventricular (RV) hypertrophy, signs consistent with human chronic mountain sickness; however, there were gender differences in the magnitude of the polycythemia and susceptibility to the fatal consequence of chronic mountain sickness. Orchiectomy and ovariectomy were performed to evaluate the role of sex hormones in the gender differences in these hypoxic responses. After 40 days of exposure to simulated high altitude (5,500 m; barometric pressure of 370 Torr and inspired Po2 of 73 Torr), both sham-gonadectomized male and female rats developed polycythemia and had increased RV peak systolic pressure and RV hypertrophy. The hematocrit was slightly but significantly higher in males than in females. Orchiectomy did not affect these hypoxic responses, although total ventricular weight was less in the castrated high-altitude rats. At high altitude, the mortality rates were 67% in the sham-operated male rats and 50% in the castrated animals. In contrast, ovariectomy aggravated the high-altitude-associated polycythemia and increased RV peak systolic pressure and RV weight compared with the sham-operated high-altitude female rats. Both sham-operated control and ovariectomized females suffered negligible mortality at high altitude. The present study demonstrated that 1) the male sex hormones play no role in the development of the excessive polycythemia, pulmonary hypertension, and RV hypertrophy during chronic hypoxic exposure or in the associated high mortality and 2) the female sex hormones suppressed both the polycythemic and cardiopulmonary responses in vivo during chronic hypoxic exposure.

Chronic exposure to simulated altitude does not increase angiogenic activity in skeletal muscle of rats.

In order to examine whether the hypoxia of high altitude increases angiogenic activity in skeletal muscle, six male Wistar rats were subjected to a simulated altitude of about 5,500 m (ambient pressure 380 mmHg) for 3 weeks, and the whole soleus, gastrocnemius, and extensor digitorum longus muscles were collected. As a result, any muscle extracts from high altitude rats did not significantly enhance the capillary growth in an in vitro angiogenesis model compared with those from sea-level rats. This appeared to confirm previous morphological studies that hypobaric-hypoxic environment did not cause the formation of new capillaries in skeletal muscles.

Role of erythrocyte deformability in the acute hypoxic pressor response in the pulmonary vasculature

To assess the importance of erythrocyte deformability in the pulmonary hypoxic pressor response (HPR) we examined whether alterations is erythrocyte deformability are related to the differences between the brisk HPR in rats vs the small HPR in hamsters, and between the HPR in low altitude rats vs high altitude rats (10 days in 10% oxygen). Deformability of the erythrocytes (RBC) was assessed by filtering equal volume of RBC suspension through Nucleopore filters (4.7 μm) using the same pressure head across the filter. The results show that during hypoxia, rat RBC become relatively nondeformable compared to hamster's RBC. This finding is consistent with a large HPR in rats but a small HPR in hamsters. Furthermore, the deformability of RBC from high altitude rats became unaffected by hypoxia and was associated with blunting in the HPR in isolated lungs from high altitude rats. The HPR in isolated lungs from low altitude rats was larger when they were perfused with blood from normal rats (= 86% increase in resistance) than when perfused with blood from high altitude rats (= 36% increase in resistance). This finding further supports the possible role of RBC deformability in HPR. Inconsistent with the importance of deformability, however, was the finding that high altitude rat lungs had a blunted HPR whether they were perfused with normal rat blood or high altitude rat blood. This may be due to restructuring of the pulmonary microvascular bed in the lung from high altitude rats. The results favor the idea that changes in erythrocyte deformability may be responsible for the difference between the HPR in low altitude rats and hamsters, and between the HPR in low and high altitude rats. We suggest that ‘obstruction’ of the capillaries by less deformable erythrocyte is another factor, besides smooth muscle contraction, responsible for the hypoxic pressor response in the pulmonary vasculature.

Reticulocytosis, increased mean red cell volume, and greater blood viscosity in altitude susceptible compared to altitude resistant rats

We have identified two strains (H and M) of Sprague-Dawley rat with markedly different susceptibilities and cardiopulmonary responses to chronic hypobaria. To further characterize factors responsible for these differing cardiopulmonary responses to chronic hypobaria, the present study examined differences in hematologic responses between the strains and assessed the contribution of differences in blood viscosity to differences in pulmonary vascular resistance. Following a 4–5 week exposure to simulated high altitude (0.5 atm), hemoglobin, hematocrit, mean red cell volume, and reticulocyte count were all increased in the susceptible H compared to the resistant M rats, whereas red blood cell counts were similar. Sea level controls manifested no differences. Blood viscosity, measured in a capillary viscometer, was 53% greater in chronically hypoxic H than in M rats, and plasma viscosities were similar. Blood from high altitude H rats increased pulmonary vascular resistance more than blood from high altitude M rats when perfused into lungs isolated from high altitude rats of either strain. In conclusion, high altitude H rats have an increased population of immature red cells, leading to a greater mean red cell volume and hematocrit than in high altitude M rats. These hematologic differences contribute to the increased blood viscosity and greater pulmonary vascular resistance of H compared to M rats after 4 weeks' high altitude exposure.

The role of pulmonary vascular responses to chronic hypoxia in the development of chronic mountain sickness in rats

A strain of Sprague-Dawley rat obtained from Hilltop Labs, Scottsdale, PA (H rats), develops more severe pulmonary hypertension, right ventricular hypertrophy, and polycythemia than a strain obtained from Madison, WI (M rats), following exposure to simulated high altitude. We sought to determine whether differences in pulmonary vascular responses to chronic hypoxia could explain the differing high altitude susceptibilities of the two strains. Vasoconstrictor responses to hypoxia and angiotensin II were tested in blood perfused lungs isolated from rats of both groups exposed to simulated high altitude (4 to 5 weeks, 0.5 atm), or from sea level controls. Pressure-flow curves, serving as an index of ‘passive’ vascular resistance, were also determined in the isolated lungs. Vasoconstrictor responses to hypoxia were blunted in high altitude rats of both the H and M strains compared to sea level controls, and the H sea level rats had blunted vasoconstrictor responses to hypoxia compared to the M sea level rats. Vasoconstrictor responses to angiotensin II were similar among the groups and were unaffected by chronic high altitude exposure. Pressure-flow curves were greater in both high altitude groups than in the sea level groups, and those of the H high altitude rats were slightly greater than those of the M high altitude rats. Thus, differences in vasoconstrictor responses to hypoxia do not explain the greater pulmonary hypertension of H high altitude rats. However, greater ‘passive’ vascular resistance, probably due to more extensive structural remodeling of pulmonary vessels, does appear to contribute to the greater pulmonary hypertension of the H rats.

Pulmonary vascular changes in young and aging rats exposed to 5,486 m altitude.

Young (YNG) and middle-aged (MA) male rats were exposed to 5,486 m for durations ranging from 1 to 42 days to determine the effect of age on the progression of polycythemia, right ventricular hypertrophy (RVH), lung vascular muscularization, and pulmonary vascular responsiveness. Other rats were exposed for 42 days at 5,486 m and were then allowed to recover at 1,520 m for periods up to 42 days. The progression and subsequent regression of polycythemia and RVH with altitude exposure were similar for YNG and MA rats. However, YNG rats exhibited vascular muscularization during the altitude exposure, characterized by hypertrophy of smooth muscle cells, whereas MA rats exhibited little or no change in vascular morphology. Lungs from both altitude-exposed YNG and MA rats exhibited blunting of acute hypoxic pulmonary vasoconstriction upon exposure to 5,486 m, with more severe blunting apparent in MA rats. Pressor responses to angiotensin II (AII) were potentiated in lungs from high altitude rats, particularly in the YNG rats, and this increased responsiveness persisted during the recovery period. A positive correlation was found in YNG rats between the degree of vascular muscularization and the pressor response to AII, suggesting that increased muscle mass was partially responsible for the potentiated AII responses. However, MA rats did not exhibit the same correlation for AII, and neither YNG nor MA rats exhibited increased responsiveness to 5-hydroxytryptamine. The results indicate that age influences the morphologic responses to altitude exposure and vascular responsiveness to AII, but does not affect the polycythemic response or the degree of RVH.

Hypoxia impairs vasodilation in the lung.

Alveolar hypoxia causes pulmonary vasoconstriction; we investigated whether hypoxia could also impair pulmonary vasodilation. We found in the isolated perfused rat lung a delay in vasodilation following agonist-induced vasoconstriction. The delay was not due to erythrocyte or plasma factors, or to alterations in base-line lung perfusion pressure. Pretreating lungs with arachidonic acid abolished hypoxic vasoconstriction, but did not influence the hypoxia-induced impairment of vasodilation after angiotensin II, bradykinin, or serotonin pressor responses. Progressive slowing of vasodilation followed angiotensin II-induced constriction as the lung oxygen tension fell progressively below 60 Torr. KCl, which is not metabolized by the lung, caused vasoconstriction; the subsequent vasodilation time was delayed during hypoxia. However, catecholamine depletion in the lungs abolished this hypoxic vasodilation delay after KCl-induced vasoconstriction. In lungs from high altitude rats, the hypoxia-induced vasodilation impairment after an angiotensin II pressor response was markedly less than it was in lungs from low altitude rats. We conclude from these studies that (a) hypoxia impairs vasodilation of rat lung vessels following constriction induced by angiotensin II, serotonin, bradykinin, or KCl, (b) hypoxia slows vasodilation following KCl-induced vasoconstriction probably by altering lung handling of norepinephrine, (c) the effect of hypoxia on vasodilation is not dependent on its constricting effect on lung vessels, (d) high altitude acclimation moderates the effect of acute hypoxia on vasodilation, and (e) the hypoxic impairment of vasodilation is possibly the result of an altered rate of dissociation of agonists from their membrane receptors on the vascular smooth muscle.