Hypoxic pulmonary steady-state diffusing capacity for CO and cardiac output in rats born at a simulated altitude of 3500 m

Summary In rats born in the low pressure chamber from sea level parents a higher hypoxic steady-state pulmonary diffusing capacity for CO was found as compared with controls of similar body weight. This difference could be explained by a difference in age or by an increase of blood O₂ capacity. There was no difference in alveolar ventilation and alveolar-arterial O₂ pressure differences, a lower cardiac output, no difference in arterial O₂ tension, no difference in arterial O₂ content but a decreased mixed-venous O₂ content as compared with control rats measured at hypoxia. A shift of the standard blood O₂ dissociation curve to the right was found in the simulated high altitude exposed rats. Calculated mixed-venous O₂ pressure was not altered in these rats; since arterial O₂ pressure was the same no difference in mean tissue capillary O₂ pressure may be presumed as compared with control animals. The results suggest that the first generation of rats exposed to simulated high altitude for their whole life is not only less adapted than animals exposed in their youth (as described in previous work) but that the ability to promote the O₂ transport in time of need in rats born in the low pressure chamber is probably even inferior to that of the controls.     

Hypoxic pulmonary steady-state diffusing capacity for CO and alveolar-arterial O2 pressure differences in growing rats after adaptation to a simulated altitude of 3500 m

Summary Steady-state pulmonary diffusing capacity for CO and alveolar-arterial O₂ pressure differences were measured at hypoxia in growing rats adapted to a simulated altitude of 3500 m. The pulmonary diffusing capacity was significantly higher and the alveolar-arterial gradients were significantly lower in the adapted animals as compared with the controls exposed to hypoxia for the first time. The increased diffusing capacity could be explained entirely by the increase of blood O₂ and CO capacity whereas the decrease of gradients might be explained by the increase of blood O₂ capacity together with an increase of the arterio-venous O₂ difference.     

Cardiac output,arterial and mixed-venous O2 saturation,and blood O2 dissociation curve in growing rats adapted to a simulated altitude of 3500 m

Summary In rats adapted to a simulated altitude of 3500 m cardiac output measured at hypoxia by the direct Fick principle was significantly lower than in the control animals (mean values 54.3 ml/min and 69.8 ml/min, resp.). The decrease of cardiac output was accompanied by an increase of arterio-venous O₂ difference and a decrease of stroke volume in the adapted rats. It is suggested that the decrease of cardiac output might be related to the increase of hematocrit. The adapted rats also showed higher arterial and mixed-venous O₂ content (both at hypoxia) and increased O₂ capacity. Arterial O₂ saturation of the animals previously exposed to simulated high altitude hypoxia was significantly higher (67.3% as against 61.2% in the controls). The standard O₂ dissociation curve showed lower oxygen affinity in the blood of the adapted animals but no physiological advantage concerning the transport of O₂ to the tissues was found. In another group of animals the Bohr factor was estimated and no difference was found between rat and human blood.     

Blood volume and body haematocrit of rats native to a simulated altitude of 3500 m

Circulating blood volume (BV) as the sum of circulating red cell volume (RCV) and plasma volume (PV) was estimated in rats native to a simulated altitude of 3500 m (“natives”), in rats born at sea level and later in life transferred to the simulated high altitude (“newcomers”), and in control sea-level rats. RCV per kg body weight (b.w.) was significantly larger in both “newcomers” and “natives” than in controls. PV per kg b.w. was in the “newcomers” insignificantly and in the “natives” significantly smaller than in the controls. BV per kg b.w. in both high altitude groups tended to be larger than in controls but the difference was not significant. Arterial haematocrit (Ahct) in the “newcomers” was significantly higher than in the controls, and in the “natives” significantly higher than in both other groups. Body haematocrit (the ratio of RCV and BV in per cent) was smaller than Ahct in all groups; this was more pronounced in the “newcomers” than in the controls and even more so in the “natives”. Apparently the haematocrit in the minute vessels of the organs of animals exposed to chronic hypoxic hypoxia increases much less than might be expected from changes of the Ahct. An attempt was made to evaluate the possible error of the more commonly used method of estimating BV, when only RCV, or only PV, is measured, and BV and its complementary fraction are calculated from arterial or venous haematocrit. When, in our results, BV was calculated from RCV and Ahct, the absolute values and also the differences between groups were somewhat underestimated. When BV was calculated from PV and Ahct, the BV itself, and particularly the differences between groups, were overestimated quite considerably. It is suggested that the only safe way to estimate BV is to measure RCV and PV separately.     

Coronary blood flow in rats native to simulated high altitude and in rats exposed to it later in life

Summary In rats exposed to a simulated high altitude of 3500 m for their whole prenatal and postnatal life a severe cardiac hypertrophy develops. In rats born and first staying 5 weeks at sea level and then being exposed to simulated high altitude, only a unilateral right cardiac hypertrophy occurs. In both groups nutritional coronary blood flow was estimated in left ventricle, right ventricle, and septum and was compared with control animals of similar age. Coronary blood flow was measured at hypoxia in all groups. At first cardiac output was determined by the Fick principle, then⁸⁶Rb was applied and the animals were killed after 55 sec. Activity of⁸⁶Rb was measured in both cardiac ventricles and septum and the fractional uptake was calculated. According to Sapirstein (1956, 1958) the distribution of⁸⁶Rb follows the distribution of cardiac output and from both these data the nutritional blood flow to the parts of the heart may be estimated.Cardiac output was similar in rats exposed to simulated high altitude later in life (newcomers) and in control animals, but it was significantly lower in rats born in the low pressure chamber (natives).Fractions of cardiac output supplying cardiac ventricles and septum in rats from both hypoxic groups were significantly higher than in control animals. In the natives they were significantly higher than in the newcomers. The fractions of cardiac output in both newcomers and natives remained significantly higher than those of the control animals, also when calculated per gram of heart tissue.Nutritional coronary blood flow (in ml/min) was higher in both ventricles and septum of the newcomers and in the right ventricle of the natives, and lower in the septum of the natives, when compared with control animals. Coronary blood flow per gram of heart tissue (in ml/min·g) was significantly higher in all cardiac parts of the newcomers, but it was about the same in all cardiac parts of the natives when compared with controls.The importance of observed changes concerning myocardial tissue oxygenation is analyzed by using Krogh's cylindrical tissue model.Presented in part at the XXVIth International Congress of Physiological Sciences, New Delhi, India, October 20–26, 1974.     

Relative organ blood flow in rats exposed to intermittent high altitude hypoxia

Summary Circulating blood volume, cardiac output and relative organ perfusion changes were studied, using the Sapirstein method of⁸⁶Rb tissue uptake, in male 75-day-old rats exposed to intermittent high altitude hypoxia (gradually up to 7000 m, 4 h daily, 5 days a week; the total number of exposures was 24).Intermittent hypobaric exposure caused a significant rise of the erythrocyte volume, whereas the plasma volume remained unchanged. The relative perfusion of the left and particularly of the right ventricular myocardium, as well as of the spleen, liver, lung, small intestine and skeletal muscle, was significantly higher. The cardiac output determined in other experimental animals similarly treated was significantly higher after 24 exposures to the intermittent high altitude hypoxia. We suggest that these changes are triggered by tissue hypoxia and a greater blood flow demand.     

Resting values of left ventricular work to coronary blood flow ratio in rats exposed to intermittent high altitude hypoxia and swimming

Summary Total hemodynamic values and left ventricular blood flow were studied using Sapirstein's method of ⁸⁶Rb uptake in female rats 24 h after a last exposure to high altitude. A simulated altitude of 1350 m was used, initial exposure being for 30 min, gradually increased by 30 min daily up to 330 min daily for 5 days a week; the total number of exposures was 32. In another animal group the hypobaric exposure was combined with swimming in water at 37 C.In both experimental groups the cardiac output and stroke volume increased, and in rats undergoing swimming the total peripheral resistance decreased as well.In the rats exposed to intermittent hypoxia only, left ventricular blood flow increased by about the same proportion as the cardiac output. The ratio of left ventricular work to coronary blood flow was significantly increased.In rats exposed to the combined influence of hypoxia and swimming, the increase in left ventricular blood flow did not match either the increase in cardiac output, or the weight gain of the left ventricle. The ventricular work to coronary blod flow ratio was the same as in controls.     

模拟高原缺氧不依赖肺动脉高压促进大鼠右心室ACE2的表达

目的： 研究模拟高原缺氧环境下大鼠心脏血管紧张素转换酶2（ACE2）的变化规律及调节因素,初步探讨ACE2与高原心脏病的关系。 方法： 在低氧环境（模拟海拔5 000 m高原、23 h/d）下饲养成年雄性 Sprague Dawley（SD）大鼠,并分为缺氧1、15、30 d组,设立平原对照组;检测各组大鼠右心室ACE2活性,同时测定右心室功能、重量指数以及肺动脉压力变化。在此基础上,我们观察了卡托普利、尼群地平灌胃对慢性缺氧30 d组大鼠右心室ACE2活性的影响。结果： 缺氧30 d 组右心室ACE2 mRNA、蛋白表达显著上调,ACE2活性明显增加,同时伴明显心室肥厚及心功能显著升高。另外,卡托普利、尼群地平虽然显著降低了肺动脉压力以及心脏功能,但对ACE2活性无显著影响。结论： 慢性高原缺氧环境促使心脏ACE2的表达与活性上调,提示ACE2可能与缺氧心脏结构功能变化有关;表达增加是缺氧环境中大鼠右心室ACE2活性升高的原因之一,而肺动脉高压可能并非是缺氧情况下ACE2活性变化的主要原因。     

Effects of exposure to a simulated altitude of 5500 m on energy metabolic pathways in rats

We examined the effect of exposure to 5500 m on three closely related metabolic pathways: anaerobic glycolysis, the pentose phosphate shunt (PPS), and fatty acid metabolism. Rats were exposed to simulated altitude of 5500 m for up to 3 months. The maximal rate of lactate production in tissue homogenates, tissue lactic acid dehydrogenase and blood lactate levels were measured to evaluate the capacity for anaerobic glycolysis. The uptake of 14C-1-palmitate, oxidation of 14C-1-palmitate to 14CO2, incorporation of 14C-1-palmitate into tissue lipids, plasma and tissue free fatty acids (FFA) levels and total lipid contents were measured to assess the magnitude of lipid metabolism. Activities of glucose-6-phosphate dehydrogenase (G-6-PD) and 6-phophogluconate dehydrogenase (6-PGD) in the PPS pathway were measured to assess the capacity to generate reducing power. Acute and chronic hypoxia did not affect most of the measurements of anaerobic glycolysis, but depressed lactate production in liver and kidney. Chronic hypoxia enhanced all aspects of lipid metabolism in liver and enhanced the uptake and oxidation to CO2 of palmitate in skeletal muscle. Chronic hypoxia did not alter the activity of the G-6-PD in any tissue studied, but the activity of 6-PGD was depressed in heart, kidney, thymus and adrenal gland. The lack of major changes in the capacities of anaerobic glycolytic pathways and the activities of the PPS dehydrogenases is consistent with the maintenance of normal aerobic metabolism in rats at 5500 m. We found no evidence that anaerobic metabolic processes were upregulated to sustain energy consumption during chronic hypoxia. On the other hand, enhanced fatty acid metabolism may spare carbohydrate for metabolic fuel under conditions of extreme hypoxic limitation.     

改进的大鼠心脏异位移植模型的应用

目的 以0no建立的大鼠心脏异位移植术为基础，并对其进行简化和改进。方法 供体为SD大鼠，受体为Wistar大鼠，将供心升主动脉和肺动脉分别与受体的腹主动脉和下腔静脉吻合。结果 50次正式实验成功率达到85％，手术时间和从心缺血时间缩短，心肌保护良好，移植心存活时间2－14d。结论 该方法方便、简单，是研究器官移植排斥反应以及筛选免疫抑制药物的一种较为理想动物模型，值得进一步推广。     

Age-related changes in parvalbumin in the heart of female rats

Changes of parvalbumin protein levels and immunolocalisation during the postnatal development of the female rat heart were investigated in order to determine if they were correlated with age-related changes in cardiac function. Hearts from newborn, 3-month-old (young), 6-month-old (young adult) and 12-month-old (adult) female Wistar rats were processed for immunohistochemical localization of parvalbumin and for Western blotting assay. Parvalbumin was detected by both methods in all age groups from newborn to 12-month-old rats. In the newborn rat heart, parvalbumin immunoreactivity did not fully fill the sarcoplasm of the cardiac myocytes and the amount of parvalbumin was low compared to the adult levels. In contrast, in 3-12-month-old rats, strong parvalbumin immunoreactivity was detected throughout the sarcoplasm of all cardiac myocytes and the amount of parvalbumin increased with increasing age (from newborn to adult). Our study indicates that an increase of parvalbumin levels in the female rat heart with increasing age may be associated with maintenance of proper relaxation of the cardiac myocytes needed to cope with the increasing workload of the heart during postnatal growth.     

EEG, ECG and oxygen concentration changes from sea level to a simulated altitude of 4000m and back to sea level

In order to describe how high altitude affects the body during a one night stay at 4000m experiments were performed in a hypobaric chamber and compared to a study on Dachstein (mountain in Austria, 2700m). Ten subjects had to perform a reaction time task at different altitudes. The EEG and ECG were recorded simultaneously. Additionally, the oxygen saturation of the blood was measured at different altitudes and the subjects filled out a Lake Louise questionnaire that describes the degree of altitude mountain sickness (AMS). After elevation from 134m to 4000m in the hypobaric chamber heart-rate increased from 68.9bpm to 81.6bpm, RMSSD (root mean square of squared differences of adjacent heart beat intervals) decreased from 54.3ms to 33.3ms, the LF/HF ratio increased from 2.5 to 3.9 and oxygen saturation decreased to 82.7% after 11h at 4000m altitude. The Lake Louise Score (LSS) reached 3.4 after one night at 4000m. EEG beta activity between 14Hz and 18Hz was attenuated at 4000m and also after return to 134m. The results indicate that the subjects were not able to adapt to 4000m within 12h in the hypobaric chamber. Even after 1h after the return to 134m all parameters are still affected from the night at 4000m altitude. ECG and EEG changes are in line with results obtained at 2700m height at Dachstein.     

Cardiac and renal distribution of ACE and ACE-2 in rats with heart failure

Congestive heart failure is often associated with impaired kidney function. Over-activation of the renin–angiotensin–aldosterone system (RAAS) contributes to avid salt and water retention in heart failure. While the expression of angiotensin converting enzyme (ACE), a key enzyme in the synthesis of angiotensin II (Ang II), is well established, the expression of angiotensin converting enzyme-2 (ACE-2), an enzyme responsible for angiotensin 1–7 generation, is largely unknown. This issue is of a special interest since angiotensin 1–7 counteracts many of the proliferative and hypertensive effects of angiotensin II. Therefore, the present study was designed to investigate the expression of both enzymes in the kidney and heart of rats with heart failure. Heart failure (CHF) was induced in male Sprague Dawley rats (n = 9) by the creation of a surgical aorto-caval fistula. Sham-operated rats served as controls (n = 8). Two weeks after surgery, the animals were sacrificed and their hearts and kidneys were harvested for assessment of cardiac remodeling and ACE and ACE-2 immunoreactivity by immunohistochemical staining. ACE immunostaining was significantly increased in the kidneys (4.34 ± 0.39% vs. 2.96 ± 0.40%, P < 0.05) and hearts (4.57 ± 0.54% vs. 2.19 ± 0.37%, P < 0.01) of CHF rats as compared with their sham controls. In a similar manner, ACE-2 immunoreactivity was also elevated in the kidneys (4.65 ± 1.17% vs. 1.75 ± 0.29%, P < 0.05) and hearts (5.48 ± 1.11% vs. 1.13 ± 0.26%, P < 0.01) of CHF rats as compared with their healthy controls. This study showed that both ACE and ACE-2 are overexpressed in the cardiac and renal tissues of animals with heart failure as compared with their sham controls. The increased expression of the beneficial ACE-2 in heart failure may serve as a compensatory response to the over-activity of the deleterious isoform, namely, angiotensin converting enzyme 1(ACE-1).     

模拟急进高原过程对清醒和麻醉状态大鼠血压和呼吸的影响

目的:为了全面地反映急进高原过程中机体的一些真实改变,本实验通过动态监测清醒和麻醉2种不同状态下大鼠血流动力学指标,旨在探讨清醒和麻醉状态大鼠在急性缺氧时血流动力学的差异,并以此进一步探讨其可能的机制。方法:实验将SD大鼠随机分为麻醉组、清醒组、5 000 m麻醉对照(A-5000-control)组、5 000m麻醉氨基胍(A-5000-AG)组、5 000 m清醒对照(C-5000-control)组和5 000 m清醒氨基胍(C-5000-AG)组。麻醉组和清醒组大鼠在低压氧舱从2 260 m开始,以2 m/s模拟急进高原5 000 m过程;其余4组均在模拟5 000 m海拔条件下进行。实验期间通过Power Lab生理记录仪实时、动态地监测整个过程中大鼠的系统动脉压(system arterial pressure,Psa)、中心静脉压(central venous pressure,CVP)、心率(heart rate,HR)和呼吸频率(breathing rate,BR)。结果:清醒组大鼠的HR和BR明显高于麻醉组,但MAP明显低于麻醉组。随着海拔的逐渐升高,清醒组和麻醉组大鼠均出现平均动脉压(mean arterial pressure,MAP)降低,且清醒组大鼠降低更为显著。另外,在5 000 m时,清醒组大鼠HR明显降低,而整个过程中2组大鼠的BR均无明显改变。静脉注射诱导型一氧化氮合酶(inducible nitric oxide synthase,i NOS)抑制剂氨基胍后,C-5000-AG组和A-5000-AG组大鼠动脉血压均明显升高,而HR和BR未见明显变化。结论:在急进高原过程中,血压和心率会明显下降,而呼吸频率变化不大。该现象可能的机制为:急性缺氧早期机体启动自我保护机制,活化i NOS,大量产生并释放NO,使血管舒张,可调节肺通气、引起血压下降;达到海拔5 000 m左右甚至更早时,机体可能出现失代偿,使心率减慢,引起血压进一步降低。由于受麻醉药物戊巴比妥钠的影响,麻醉状态的大鼠血压下降出现得较为迟缓,而清醒大鼠对急进高原性低氧反应迅速,能够更真实全面地反映急进高原过程中低氧引起的血流动力学改变。     

Plasma soluble HLA-G levels in a cohort of heart failure patients exposed to chemicals

Heart failure (HF) is a syndrome caused by structural and/or functional cardiac abnormalities, resulting in a reduced cardiac output and/or elevated intracardiac pressures. Several studies reported a crucial role of immune activation and inflammation in the chronic heart failure (HF) pathogenesis, suggesting that pro-inflammatory and anti-inflammatory mediators could be predictive markers of the HF development and/or progression. Human Leukocyte Antigen-G (HLA-G), a tolerogenic and anti-inflammatory class I non-classical major histocompatibility complex molecule, was reported to be upregulated in patients diagnosed with HF, suggesting a tentative to regulate the inflammatory condition. We evaluated soluble (s)HLA-G plasmatic levels in patients with stable chronic heart failure at baseline visit and after 6 and 12 months. The 14 bp Insertion/Deletion polymorphisms of the HLA-G gene was also analyzed. We showed that in HF subjects, sHLA-G levels were higher in NYHA class II and III subjects (mild-severe symptoms) (6.11 ± 1.15 ng/ml; 8.25 ± 2.27 ng/ml, respectively) in comparison with NYHA class I subjects (no symptoms) (2.35 ± 0.43 ng/ml) (I vs II: p = 0.0156; I vs III: p = 0.0122). Moreover, the exposure to chemicals seems to affect sHLA-G levels, with higher sHLA-G levels in exposed patients (3.36 ± 5.12 ng/ml) in comparison with unexposed subjects (2.01 ± 2.84 ng/ml). The HLA-G 3′UTR 14 bp INS/DEL polymorphism correlated with sHLA-G, with the 14 bp INS/INS genotype associated with higher sHLA-G levels during the 12 months follow-up in unexposed subjects (p = 0.008). In conclusion, these results support a correlation between sHLA-G levels, genetics and HF disease in presence of work chemical exposition.     

The growth of a primary tumor (sarcoma M-1) during the training of rats to hypoxia in the low pressure chamber

Summary The effect of hypoxic training of rats on the growth of transplantable tumors (sarcoma M-1) was studied in experiments involving 161 animals. In training the rats at an altitude of 6,000 m, the tumor growth was delayed by 18.6 per cent. With the rise of the training altitude there is a progressively increasing development of the tumor growth-inhibiting mechanisms. At the maximal altitude of training (10,750m) the tumor growth inhibition reached 57.5%.(Presented by Active Member, Academy of Medical Sciences, USSR, V. V. Parin) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 51, No. 6, pp. 73–76, June, 1961     

Distribution of acetylcholinesterase activity in the rat embryonic heart with reference to HNK-1 immunoreactivity in the conduction tissue

Acetylcholinesterase (AChE) activity was topographically investigated in the presumptive cardiac conduction tissue regions visualized by HNK-1 immunoreactivity in rat embryos, and AChE-positive cells were examined with the electron microscope. On embryonic day (ED) 14.5, when HNK-1 was most intensely visualized, AChE activity could not be detected enzyme-histochemically in the conduction tissue regions, except in the ventricular trabeculae and part of the AV node. On ED 16.5, however, the AChE activity was clearly demonstrated in some parts of the developing conduction tissue. One exception was the AV node region, where an AChE-positive area was in close proximity to an area showing HNK-1 immunoreactivity but did not overlap. Furthermore, AChE activity was demonstrated predominantly in the ventricular trabeculae, including cardiac myocytes, but was rather weak in the atrium. With the electron microscope, AChE reaction products were observed predominantly intracellulary in both developing conduction tissue cells and developing ordinary myocytes, and no reactivity was found in neuronal components. From ED 18.5 until birth, both AChE activity and HNK-1 immunoreactivity faded away in the conduction tissue. Thus, transient AChE activity in the embryonic heart seems to be different from the developing adult form and may be related to a morphogenetic function in embryonic tissues, as proposed by other authors.     

Oxidative stress induced by intermittent exposure at a simulated altitude of 4000 m decreases mitochondrial superoxide dismutase content in soleus muscle of rats

The effects were examined of 6-month intermittent hypobaric (4000 m) exposure on the antioxidant enzyme systems in soleus and tibialis muscles of rats. At the end of the 6-month experimental exposure, the six rats in both the exposed group and the control group were sacrificed. Immunoreactive mitochondrial superoxide dismutase (Mn-SOD) contents were measured as well as the activities of antioxidant enzymes [Mn-SOD, cytosolic SOD (Cu,Zn-SOD), catalase (CAT), and glutathione peroxidase (GPX)]. Thiobarbituric acid-reactive substances (TBARS) were also determined as an indicator of lipid peroxidation. The high altitude exposure resulted in a marked increase in TBARS content in soleus muscle, suggesting increased levels of oxygen free radicals. Conversely, significant decreases in both Mn-SOD content and activity in solens muscle were oted affer exposure. Such trends were not noticed in tibialis muscle. On the other hand, no significant changes in Cu,Zn-SOD, CAT, or GPX were observed in either muscle. These results suggested that the increases in lipid peroxidation were most probably a result of decreased Mn-SOD function which was more depressed in oxidative than in glycolytic muscle.