Manifestation of oxidative stress in the pathogenesis of arterial hypertension in ISIAH rats.

The ISIAH rat strain with stress-sensitive arterial hypertension was intentionally selected to study the role of stress as a factor in the development of arterial hypertension. This study aimed to determine the role of reactive oxygen and nitrogen species (ROS and RNS) in the pathogenesis of hypertension in ISIAH rats. The nitric oxide concentrations measured by EPR were found to be significantly higher for hypertensive ISIAH rats compared with that for normotensive Wistar rats in both the aortic wall (2 times) and cerebellum (1.5 times). The activity of superoxide dismutase measured in the blood of ISIAH rats was found to be about 1.5 times lower compared with that of Wistar rats. These data support the suggestion that ROS and RNS, including superoxide radicals and nitric oxide, may play an important role in development of stress-induced hypertension in ISIAH rats. The tissue content of reduced thiols has been considered as a marker of oxidative damage. To study the tissue oxidative status we used an EPR method for quantitative determination of SH groups. The concentration of reduced thiols in the blood of ISIAH rats was much lower than that in Wistar rats (0.6 +/- 0.05 and 1.57 +/- 0.1 mM, respectively).     

Features of the behavior of the rat with hereditarily determined arterial hypertension

The behaviour of spontaneously hypertensive rats (SHR strain), rats with inherited stress induced arterial hypertension (ISIAH, a new developed strain), and of their normotensive Wistar progenitors was studied. The open-field arena and a device for measuring the total activity in the home cage were used in the behavioural studies. The SHR were much more active in the open--field and home cage tests than the Wistar and ISIAH rats. The basal locomotor activity of the ISIAH strain was lower than that of the Wistar rats, but the ISIAH strain had an index of behavioural reactivity 2.7 fold higher than the Wistar or SHR strains. These behavioural characteristics corresponded to the hypertension patterns of the strains compared. Enhanced spontaneous locomotion of the SHR rats was associated with spontaneous increase in arterial blood pressure. The ISIAH rats showed low spontaneous locomotor activity, but high behavioural and blood pressure reactivity under conditions of mild emotional stress.     

Chronic stress and forming of psychoemotional state during prepubertal period in rats

Immediate and long-lasting effects of chronic stress during prepubertal period (21-32 postnatal days) on anxiety- and depression-related behavior were studied in Wistar and ISIAH (inherited stress-induced arterial hypertension) rats. Significant interstrain differences were found. Both juvenile and adult ISIAH rats were less anxious in the elevated plus-maze and less depressed in the forced swimming test. Immediate effects of the prepubertal stress were similar in both rat strains and depended on the type of stimulation. Long-lasting effects were genotype-dependent. Chronic prepubertal handling exerted an anxiolytic effect in young ISIAH and Wistar rats and adult Wistar rats. Immediate anxiogenic effect of prepubertal unpredictable stress was preserved only in adult ISIAH rats. Depression-related behavior was intensified by the unpredictable stress in young animals, whereas the long-lasting effect was observed only in adult hypertensive rats.     

Typological features of behavior and memory in Norway rats selected for absence of aggression toward a man

Features of behavior and retrieval of passive conditioned avoidance on a new and forgotten stimuli were compared in Wistar rats and Norway rats bred for the absence of aggression toward a man. As distinct from white rats, grey rats were characterized by low anxiety and high locomotor exploratory activity in the elevated plus maze and dark-light chamber. Norway rats demonstrated better avoidance performance with active defensive behavioral strategy than Wistar rats. Latent inhibition during conditioning with a previously forgotten situational stimulus was the same in both rat strains. The results are discussed in terms of the use of grey rats as a model for an in-depth analysis of the mechanisms of memory optimization.     

Effect of chronic stress during prepubertal period of life on development of inherited arterial hypertension

The immediate and long-lasting effects of environmental stress during prepubertal life on arterial blood pressure (AP) were studied in rats with inherited stress-induced arterial hypertension (ISIAH) and normotensive Wistar rats. Two models of chronic stress (the 21st-32nd postnatal days) were used: repeated handling and unpredictable stress of daily exposures to a variety of mild physical or psychoemotional stressors. Chronic prepubertal stress did not affect the basal or stress-induced AP levels in young or adult Wistar rats. In ISIAH rats, chronic stress during the early phase of hypertension development did not accelerate its formation and did not augment its manifestation in adults. Moreover, the basal AP was decreased in young and adult ISIAH rats exposed to prepubertal stress as compared to the age-matched controls. AP elevation under acute stress conditions was lower in young ISIAH rats exposed to unpredictable stress. No long-lasting effect of prepubertal stress on acute stress-induced AP elevation in adults was found. The conclusion was drawn that moderate physical and psychoemotional training at prehypertensive stage can positively affect the development of inherited arterial hypertension.     

The effect of social isolation during juvenile period on the sexual arousal in adult rats

The effects of chronic postweaning social isolation combined with subsequent resocialization on the sexual arousal were studied in male rats with inherited stress-induced arterial hypertension (ISIAH strain) and in Wistar rats. Young males were isolated on the Day 21 of postnatal life for 6 weeks. Then they were kept in groups of 5. 4-month males underwent the partition test: a receptive female was introduced into the male's cage, but the male and the female were separated by a transparent partition. The number of approaches to the partition and total time spent near the partition during the test served as an index of sexual motivation. Hormonal component of sexual arousal was estimated by measuring plasma testosterone level. No interstrain differences in magnitude of the sexual arousal were observed. However, chronic social isolation during juvenile period caused a genotype-dependent diminution of sexual motivation in the adult male rats of both strains. The decrease of sexual motivation in ISIAH rats was more pronounced as compared to Wistar rats. Moreover, the social isolation during the juvenile period completely abolished the female-induced rise in plasma testosterone in ISIAH and Wistar male rats. Plasma corticosterone level was increased during the period of sexual arousal, but this rise of corticosterone was not affected by the social conditions during postweaning period.     

Analysis of polymorphic DNA markers cosegregation with arterial blood pressure in ISIAH hypertensive rats]

A search for DNA markers of hereditary arterial hypertension in ISIAH rats was performed by means of contemporary molecular genetic approaches. The backcross rat population used for the analyses was derived from a cross of the Wistar x ISIAH F1 progeny with the Wistar rats. Hybridization of the HaeIII-digested DNA samples with the (CAC)5 microsatellite probe revealed cosegregation of the basal arterial pressure value with the 4.8-kb polymorphic DNA fragment. Examination of the DNA polymorphism by means of polymerase chain reaction with arbitrary primers showed an association of the 700-bp polymorphic DNA fragment with the increase of arterial blood pressure under conditions of emotional stress.     

Renal and endocrine changes in rats with inherited stress-induced arterial hypertension (ISIAH)

Hypertensive inbred rats (ISIAH; inherited stress-induced arterial hypertension) present with baseline hypertension (>170 mmHg in adult rats), but attain substantially higher values upon mild emotional stress. We aimed to characterize key parameters related to hypertension in ISIAH. Kidneys, adrenals, and systemic endocrine parameters were studied in ISIAH of different ages and compared to normotensive Wistar albino Glaxo (WAG) rats. Native organs were obtained for Western and PCR analysis. Perfusion-fixed organs were prepared for histopathology and quantitative histochemistry. Plasma renin and adrenal hormones were measured. Renal morphology was unaltered in ISIAH. The hypothalamic-pituitary-adrenocortical (HPA) axis was constitutively upregulated with enlarged adrenal cortices and enhanced plasma corticosterone levels. Plasma renin activity was not different between groups, whereas aldosterone levels were in part reduced. Juxtaglomerular NO synthase type 1, cyclooxygenase type 2, and renin expression were significantly reduced, whereas tubular gene products related to sodium transport (bumetanide-sensitive Na, K, 2Cl cotransporter type 2; thiazide-sensitive Na, Cl cotransporter; epithelial Na channel-α; 11β-hydroxysteroid dehydrogenase type 2) were increased. These data suggest enhanced volume conservation by the kidney. Our data define ISIAH as an attractive model for the renal components determining salt and water homeostasis in hypertension. The specific condition of a basally stimulated HPA axis is highlighted, including the option to study effects superimposed by emotional stress.     

Function of hypothalamic-pituitary-adrenocortical system in hypertensive rats of ISIAH strain

Functional activity of hypothalamic-pituitary-adrenocortical axis has been studied under control and restraint stress conditions in rats with inherited stress-sensitive arterial hypertension (ISIAH strain) and in normotensive WAG (Wistar Albino Glaxo) strain. The levels of hypothalamic CRH-mRNA (in control and 2 hrs stress), pituitary and plasma ACTH and plasma corticosterone (in control and after 5, 15 or 30 min of restraint stress), were evaluated. Hypothalamic CRH-mRNA level was found to be approximately the same in the control rats of both strains. In control conditions, the pituitary and plasma ACTH content in ISIAH rats was significantly lower whereas the corticosterone level in the plasma differed from each other in both strain. The restraint stress resulted in a statistically significant increase of the CRH-mRNA in ISIAH rats and not in the WAG rats. Moreover, in spite of the lower ACTH level in stressed ISIAH rats, the corticosterone blood plasma concentration in hypetensive rats was significantly higher. The data obtained confirm the idea that the stress-dependent hypertension might be related to an enhanced sensitivity of the main endocrine links involved in the stress response organization.     

Expression of the pro-opiomelanocortin gene in rats with inherited arterial hypertension]

Expression of pro-opiomelanocortin (POMC) gene in pituitary of rats from newly developed hypertensive strain (ISIAH strain) was studied by dot hybridization. The pUC8 plasmid containing 900 base pair (bp) segment or the human POMC gene corresponding to the major portion of the 3'-nontranslated mRNA region and 60 bp coding for the signal peptide, was used as a probe for hybridization. It was found that the expression of the POMC in pituitary of the hypertensive JSJAH rats was more than 3-fold gene lower as compared to normotensive Wistar strain. The latter is the original strain from which the ISIAH rats were bred. The mechanism of this phenomenon and its possible relation to the arterial hypertension are discussed.     

Behavioral characterictics of ISIAH rat strain

This work was performed in hypertensive ISIAH and normotensive WAG rat strains. The latter was used as a control group. The general evaluation of behavior of the ISIAH and WAG rats tested in the open field, in the light-dark test, in the sound stress test, and in the fatigue test showed that the motor and exploratory activity provoked by an unfamiliar environment was much higher in the ISIAH rats as compared to the control WAG strain. Spontaneous locomotor activity of the ISIAH rats in the home cage was significantly lower as compared to the control WAG strain. This finding suggests that the ISIAH rats are hyperreactive in an novel environment. It is concluded that the hypertensive ISIAH rats are significantly different from the control WAG rats not only in the arterial blood pressure level, but also in behavioral patterns.     

Characteristics of glucocorticoid receptor gene expression in spontaneously hypertensive rat strain

In ISIAH rat strain with stress-sensitive form of hypertension, the expression level of glucocorticoid receptor (GR) gene has been evaluated in hippocampus, hypothalamus and pituitary under basal and 2-hr restraint stress conditions. Corticosterone (CS) level in peripheral blood was also evaluated. Normotensive WAG strain was used as a control. Under basal condition, there were no interstrain differences in GR-mRNA level in any brain region under study. However, under stress condition, ISIAH rats demonstrated a significant fall of GR-mRNA in hippocampus and increase the pituitary gland as compared to basal level. On the contrary, no differences with basal level were found in stressed WAG rats. CS concentration in blood was nearly the same in nonstressed WAG and ISIAH rats. Stress influence led to a marked increase of CS in both strains. However CS level was significantly higher in stressed ISIAH rats than in stressed WAG group.     

Blood pressure increase in foster mothers in ISIAH and Wistar rats: effect of reciprocal cross-fostering

Maternal arterial blood pressure (ABP) of hypertensive (ISIAH) and normotensive (Wistar) rats rearing their natural litter or litter of opposite strain, was assessed when the pups were 1.5-, 3- and 4-month old and compared with the ABP of these rats prior to their mating. The ABP was increased in both breeds of rat strains for either cross-fostered or infostered pups. No ABP changes were observed in control rats.     

Genetic correlation between the arterial pressure response during emotional stress and the alpha1-adrenoreceptor concentration in brain regions]

Arterial blood pressure reactivity to the emotional stress and brain alpha 1-adrenoreceptors concentrations were studied in hypertensive (ISIAH strain) and normotensive (Wistar strain) rats and their F1 and F2 hybrids. Significant correlations between the stress-induced increase in the arterial blood pressure and the amount of alpha 1-adrenoreceptors in hypothalamus (+0.46) and medulla (+0.38) were found in the F2. This cosegregation may point to the significant role of genetically determined peculiarities of expression of alpha 1-adrenoreceptors in brain regions during pathogenesis of arterial hypertension in the ISIAH strain.     

Expression of the renin angiotensin system genes in the kidney and heart of ISIAH hypertensive rats

The content of mRNA of renin-angiotensin system (RAS) genes was measured in the kidney and heart of hypertensive ISIAH and normotensive WAG rats using the real-time PCR. A statistically significant decrease in the mRNA level of RAS genes was registered in the kidney of ISIAH rats, including Ren (by 45%), Ace (43%), AT1A (34%), COX-2 (50%). The level of myocardial expression of AT1A decreased by 28% while Ace expression increased by 80%. These results suggest reduction of renal RAS basal activity in the hypertensive ISIAH rats, and therefore this strain of rats may be referred to the group of models of low-renin hypertension. The ISIAH rats were also characterized by a two-fold increase in the connective tissue sodium concentration and also by a small (but statistically significant) increase in plasma sodium concentration (139 ± 0.3 mmol/l versus 136 ± 0.25 mmol/l in WAG rats). These results together with a tendency to a decrease of plasma aldosterone level also support existence of a classical low-renin hypertension in the ISIAH rats. It is suggested that altered function of renal ion channels represents a basis for the development of low renin hypertension in the ISIAH rats. In addition, impairments in renal system of NO synthesis may also contribute to the pathogenesis of arterial hypertension in the ISIAH rats.     

Endocrine-metabolic relations in rats with genetic arterial hypertension

Levels of triiodothyronine, thyroxine, insulin, glucose and free fatty acids in the blood; contents of adrenaline, noradrenaline in adrenals and glycogen in the liver; activity of phenylethanolamine-N-methyltransferase in adrenals, hexokinase and glucose-6-phosphatase in the liver were studied in male Wistar rats and rats with inherited stress-induced arterial hypertension /ISIAH/. It was found that genetically caused rise of hypophyseal-thyroid systems activity in ISIAH-rats leads to a decrease of insulin blood level, activation of lipolysis and breach of glucose tolerance.     

Oxidative stress in pathogenesis of arterial hypertension in ISIAH rats]

The NO-synthase activity and the rate of NO production in the rat aortic wall and cerebellum were 2-1.5-fold higher in the ISIAH rats than in normotensive WAG rat strain. In contrast, the superoxide dismutase (SOD) activity was significantly reduced in the ISIAH rats. The blood level of reduced thiols was also much lower in the ISIAH rats. The findings suggest that oxidative stress may play a significant role in pathogenesis of stress-sensitive hypertension in the ISIAH rat strain.     

Genetic loci for spleen weight and blood pressure in ISIAH rats with inherited stress-induced arterial hypertension

Recently, the important role of the spleen’s function in hypertension development was demonstrated. In this study, the genetic control of absolute and relative spleen weight was investigated to reveal the genetic loci common for spleen traits and for arterial blood pressure at rest and under the emotional stress conditions in ISIAH rats with inherited stress-induced arterial hypertension. The search for genetic loci for absolute and relative spleen weight was performed on 6-month-old F₂ (ISIAH × WAG) hybrid males derived from a cross of hypertensive ISIAH and normotensive WAG rats. One significant QTL mapped on chromosome 1 and 5 suggestive loci were found for relative spleen weight. Four suggestive loci were detected for absolute spleen weight. All detected loci were novel. The significant QTL on chromosome 1 was common for relative spleen weight and arterial blood pressure at rest and under the emotional stress conditions in ISIAH rats. The results suggest that the manifestation of the stress-sensitive arterial hypertension in ISIAH rats may be related to the changes in genetic control of the spleen function.     

A search for genetic loci responsible for emotional stress-induced arterial hypertension in the ISIAH rats

Hypertension is a widespread human disease caused by a complex interaction of a series of the genetic factors with both each other and the environmental conditions. In this study we aimed at determining the candidate genetic loci responsible for hypertension in the ISIAH rats and studying the dynamics of the relevant genetic and physiological mechanisms in rat ontogeny. The candidate genetic loci were identified from association of the microsatellite markers linked to these loci with arterial hypertension in rat F2 hybrids exposed to stress. Two populations of F2 hybrids of different age (3-4 and 6 months) were obtained by crossing hypertensive ISIAH and normotensive WAG rats. We present the results of cosegregation analysis for the following loci: the gene for the Na+, K(+)-ATPase alpha 1 subunit isoform (Atp1a1), the endothelin-2 gene (Edn2), the low affinity nerve growth factor receptor gene (Lngfr), and a region of chromosome 10 marked with the D10Rat58 microsatellile located 3 cM away of the aldolase C gene (AldC). The results obtained allowed us to localize the genes responsible for the stress-induced arterial hypertension in the ISIAH rats to the Atp1a1 locus (P < 0.05), chromosome 2 and to the Lngfr gene locus (P < 0.05), chromosome 10. The association of hypertensive status with the Lngfr gene was found only in young ISIAH rats whereas in adult rats of this line, hypertension was associated with the Atp1a1 locus.     

The electrical resistance of the lungs,intercostal muscles,and kidneys in hypertensive ISIAH rats

A comparative study of the bioelectrical impedance of normotensive and hypertensive animal tissues was carried out. It was found that the electrical resistance of kidneys, lungs, and intercostal muscles in rats with arterial hypertension (ISIAH strain) was significantly lower than in normotensive Wistar rats, which indicates that the volume of circulating blood and the total amount of fluid were increased in animals with arterial hypertension. The fact that the resistance of the conducting medium is decreased in arterial hypertension should be taken into account in the analysis of cardioelectric potentials on the body surface and electrocardiograms in conventional leads, as well as for the purposes of the development of heterogeneous torso models and for verification of recovery algorithms for electrical properties of chest tissues.