Electric muscle stimulation in the hind leg of the spontaneously hypertensive rat induces a long-lasting fall in blood pressure

The influence of prolonged low-frequency, low-intensity electric stimulation of the gastrocnemius muscle or of the biceps femoris muscle on blood pressure and heart rate was investigated in unanaesthetized, spontaneously hypertensive rats (SHR). In both groups, elevations of blood pressure and heart rate were elicited during the 60 min of muscle stimulation. After cessation of the stimulation, a depressor response developed within 60 min. Thirty to sixty minutes post-stimulation the fall in blood pressure was 19 ± 3 and 17 ± 4 mmHg, respectively (mean ± SE) compared with controls. In both groups, the depressor response lasted for over 5 h. In addition, the gastrocnemius-stimulated animals also developed a post-stimulatory bradycardia. In one group of SHR the sciatic nerve was anaesthetized with bupivacaine. The arousal response during stimulation was similar to that in the other groups, but after termination of stimulation blood pressure returned to the control level without any further drop. To investigate further the neurotransmitters involved, one group of gastrocnemius-stimulated SHR was given naloxone by infusion during the stimulation. A modest post-stimulatory blood pressure fall also occurred in this group, but it lasted only 90 min. Another SHR group was pre-treated with parachlorophenylalanine, a serotonin synthesis blocker, which completely abolished the post-stimulatory depressor response. These results indicate that prolonged muscle stimulation gives rise to a post-stimulatory long-lasting drop in blood pressure and that this response is mediated by somatic nerve afferents. Involvement of the endorphin and serotonin systems is also suggested.     

Acute mental stress but not enforced muscle activity transiently increases natural cytotoxicity in spontaneously hypertensive rats

The influence of acute mental stress and the effect of electrically induced skeletal muscle contractions on natural cytotoxicity in vivo was investigated in spontaneously hypertensive rats. Natural cytotoxicity in vivo was measured as the clearance of injected ⁵¹Cr-labelled YAC-1 lymphoma cells from the lungs, which are specifically lysed by natural killer cells. The mental stress consisted of an air jet directed towards the animals in their cage for 25 min. During the mental stress there was a significant increase in natural cytotoxicity. Thus, retained radioactivity in the lungs was decreased to 74±6% of the control levels which was set to 100% (P<0.01). This augmentation of YAC-1-cell clearance could be blocked with the β-adrenergic receptor antagonist Timolol. Two hours after termination of the air stress, in vivo cytotoxicity had returned to control levels. In contrast, acute physical stress, consisting of electrically induced muscle contractions for 60 min, had no significant effects on in vivo cytotoxicity, either during the stimulation or 1, 2 or 24 h after the stimulation. Further, significantly increased plasma levels of adrenaline were seen after the air jet stress, but not after muscle stimulation. There were no significant changes in plasma noradrenaline levels either after air stress or muscle stimulation. These results indicate that changes in in vivo cytotoxicity after mild mental stress are dependent on increased plasma catecholamine levels while acute physical stress, without changes in catecholamine levels, does not influence in vivo cytotoxicity.     

Na+, K+-pump activity and skeletal muscle contractile deficits in the spontaneously hypertensive rat

Skeletal muscles in an animal model of genetic hypertension (the spontaneously hypertensive rat, SHR) exhibit significant deficits in contractile performance. These deficits appear to be unrelated to the rise in blood pressure. Slow-twitch soleus muscles show a decrease in specific muscle tension and a reduced resistance to muscle fatigue during prolonged contractile activity. We tested the hypothesis that the reduced fatigue resistance occurs as a consequence of an impaired ability to maintain or restore Na⁺ and K⁺ balance across the sarcolemma during repeated contractions. This may result from a genetically based increase in the Na⁺ permeability of SHR muscles, coupled with a reduction Na⁺, K⁺ pump capacity as the animals mature. Soleus muscles in adult SHR exhibit a significant increase in intracellular Na⁺ content and a significant decrease in intracellular K⁺ content at rest. ⁸⁶Rb⁺ uptake in Na⁺-loaded hypertensive muscles is 45% less than predicted from the number of ouabain-binding sites available. Activation of Na⁺, K⁺ pumps using adrenaline or insulin produces a significantly smaller hyperpolarization in hypertensive soleus than in control muscles. Control soleus muscles are hyperpolarized for at least 10 min after a 4 min period of high-frequency activity, but hypertensive soleus muscles remain at resting polarity. Nonetheless, the number of ouabain-binding sites in hypertensive muscles is significantly greater than in control soleus, and binding affinities are similar. This apparent deficit in pump capacity might lead to a greater and more prolonged increase in extracellular K⁺ during repetitive contractions, and an associated decline in tension. Recently, we have been able to prevent the abnormal decrease in hypertensive soleus fatigue resistance by long-term treatment (8 weeks) with the Ca²⁺ blocker amlodipine. The therapy prevented or reversed the contractile deficits, but did not restore the responsiveness of the Na⁺, K⁺ pump to hormonal stimulation. The current data suggest that both a reduction in Na⁺, K⁺-pump capacity and changes in Ca²⁺ distribution play a role in the development of contractile deficits in hypertensive muscles.     

Effect of different frequencies of music on blood pressure regulation in spontaneously hypertensive rats

The effect of different frequencies of music on brain function was investigated through measurement of blood pressure in spontaneously hypertensive rats (SHR). Previous studies indicated that exposure to Mozart's music (K. 205) leads to increased calcium/calmodulin-dependent dopamine synthesis in the brain, and that the subsequent increase in dopamine reduces blood pressure via D(2) receptors. The present study demonstrated that the blood pressure-reducing response was dependent on the frequency, and was markedly greater at 4 k-16 kHz compared with lower frequencies. These findings suggest that music containing high-frequency sounds stimulates dopamine synthesis, and might thereby regulate and/or affect various brain functions.     

In vitro proliferation of aortic smooth muscle cells from spontaneously hypertensive and normotensive rats

The characteristics and proliferation of aortic smooth muscle cells (SMC) from spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto (WKY) rats were studied in culture. Smooth muscle cells were isolated from the tunica media of the thoracic aorta by an explant method. Immunofluorescence microscopy showed that 93-95 per cent of cells were positively labelled with antibodies raised against smooth muscle actin, indicating that these were smooth muscle cells. The proliferative activity was compared between aortic smooth muscle cells from hypertensive and normotensive rats in culture by thymidine incorporation and cell number determinations. The results demonstrate that aortic smooth muscle cells from hypertensive rats grew faster than those from normotensive rats in culture. The increased proliferative activity of cultured aortic smooth muscle cells from hypertensive rats was detectable even when they were cultured in a chemically defined serum-free medium. These data have shown that an increased proliferative activity of aortic smooth muscle cells from hypertensive rats can occur in culture conditions without the influence of arterial pressure or other stimuli as in intact animals. The mechanisms underlying the accelerated proliferative activity of aortic smooth muscle cells from genetically hypertensive rats in vitro remain to be determined.     

Role of different serotonergic receptors in the long-lasting blood pressure depression following muscle stimulation in the spontaneously hypertensive rat

In a previous study prolonged low-frequency muscle stimulation in the hind leg of the spontaneously hypertensive rat (SHR) was shown to induce a reduction in blood pressure (about 15 mmHg) that lasted for many hours. We showed in that study that endorphin and serotonin systems were involved. In the present study drugs with selective affinity for the serotonin (5-HT) receptors were used to analyse further the involvement of different serotonin systems. In one group of SHR, a prestimulatory dose of metitepine maleate (a 5-HT1 and 5-HT2 receptor antagonist) completely abolished the post-stimulatory depressor response. The long-lasting depressor response was still present, although less pronounced, after a bolus dose of the 5-HT2 blocking agent ritanserin (R 55667) at the start of stimulation. The 5-HT3 receptor antagonist ICS 205-930 did not influence the response at all, nor did the selective 5-HT1a receptor agonist 8-OH-DPAT enhance the depressor response. These results indicate that the reduction in blood pressure after muscle stimulation is mainly mediated by the 5-HT1 receptor.     

Noradrenergic hyperinnervation may inhibit necrosis of coronary arterial smooth muscle cells in stroke-prone spontaneously hypertensive rats

Noradrenergic (NA) nerve fibre distribution and vascular smooth muscle morphology were investigated in the coronary artery of stroke-prone spontaneously hypertensive rats (SHRSP). Fluorescent NA nerve fibres of SHRSP aged 10, 30, 60, 90 and 180 days were examined by the glyoxylic acid method and compared with those of age-matched normotensive Wistar Kyoto (WKY) rats. The distribution densities of NA nerve fibres were measured by quantitative image analysis using the Interactive Bildanalyse System. The densities of NA nerve fibres of the left coronary artery of SHRSP were significantly higher than those of WKY rats at all ages examined. NA hyperinnervation in the coronary artery of SHRSP may be caused by the hyperfunction of the stellate ganglia which innervate the coronary arteries. Scanning electron microscopy observations showed that the surface of smooth muscle cells of the left coronary artery in SHRSP was smooth and similar to that of WKY rats at 120 days of age, but was slightly modified by more invaginations and projections than that in WKY rats at 180 days of age. No necrotic cells, however, were found in SHRSP. By transmission electron microscopy the smooth muscle cells in SHRSP were shown to be irregular in profile with deep indentations of the plasma membrane and surrounded by many layers of basal laminalike material, but no necrotic cells were found. We suggest that NA hyperinnervation protects the vascular smooth muscle cells from necrosis in the coronary artery of SHRSP by a trophic effect mediated by NA nerve fibres.     

Effects of conditioning vibratory stimulation on pain threshold of the human tooth

The effect of mechanical vibratory stimulation on the human tooth pain threshold has been studied in 4 healthy naive subjects. Vibration at 10 and 100 Hz was applied to a maxillary incisor and the electrical pain threshold was measured in an adjacent tooth. During the conditioning stimulation the pain threshold increased 1.2–1.8 times the unconditioned values. The present study indicates that mechanical vibratory stimulation applied to human teeth elevates the pain threshold in adjacent teeth.     

The working memory capabilities of the spontaneously hypertensive rat

This study addresses the working memory capabilities of the male spontaneously hypertensive rat (SHR) strain as compared to the normotensive inbred strain, Wistar Kyoto (WKY), and the out bred Sprague Dawley (SD) rat as a normal control. The objective was to use two working memory tasks in the water maze with different strategic demands: forced alternation (FA) which allows the use of either an allocentric ("place") or egocentric ("response") localisation strategy and delayed matching-to-place (DMP) which requires an allocentric strategy. In the FA task, SHR reached criterion at the same rate as WKY and SD controls and were impaired to the same extent as WKY at the long (1 h) delay. Furthermore, both SHR and WKY were impaired relative to SD when the memory load was increased through the use of massed trials. In the DMP task, the performance of SHR did not differ from that of either of the control strains, either during training or in response to delay. These findings do not provide evidence of short-term memory impairments in the SHR, which is a commonly-used animal model of Attention Deficit Hyperactivity Disorder (ADHD) in humans.     

Electric muscle stimulation in the spontaneously hypertensive rat induces a post-stimulatory reduction in activity: role of different opioid receptors

We have previously shown that prolonged low-frequency muscle stimulation, inducing contractions of the gastrocnemius muscle, in conscious spontaneously hypertensive rats leads to an opioid-mediated post-stimulatory reduction in blood pressure and analgesia. In the present study we investigated whether muscle stimulation would also induce a post-stimulatory reduction in behavioural activity in the spontaneously hypertensive rats. Selective opioid receptor antagonists were used to analyse the involvement of endogenous opioids. Muscle stimulation, lasting 60 min, induced a post-stimulatory sedation that outlasted the stimulation for hours. Sniffing, locomotor activity and total behavioural activity were significantly reduced. The post-stimulatory reduction in activity was reversed back to control levels by a high dose of naloxone (15 mg kg-1 i.v.). The selective mu-receptor antagonist beta-funaltrexamine, given intracerebroventricularly before stimulation, did not influence the development of the post-stimulatory drop in activity. The delta-receptor antagonist ICI 154,129 had no effect at all on the already developed sedation, whereas MR 2266 BS, a kappa-receptor antagonist (3 mg kg-1 i.v.), completely reversed the drop in activity. These results show that muscle stimulation gives rise to an opioid-mediated post-stimulatory reduction in activity in spontaneously hypertensive rats. The results also indicate the involvement of the opioid kappa-receptor in the behavioural response.     

Experimental muscle pain decreases the frequency threshold of electrically elicited muscle cramps

This study in humans tested the hypothesis that nociceptive muscle afferent input facilitates the occurrence of muscle cramps. In 13 healthy adults, muscle cramps were experimentally induced in the foot by stimulating the tibialis posterior nerve at the ankle with 2-s bursts of stimuli separated by 30 s, with stimulation frequency increasing by 2-Hz increments from 10 Hz until the cramp appeared. The minimum stimulation frequency that induced the cramp was defined “cramp frequency threshold”. In 2 days, elicitation of the cramp was performed in the two-feet with and without (baseline condition) injection of hypertonic (painful condition) or isotonic (control condition) saline into the deep midportion of the flexor hallucis brevis muscle, from where surface EMG signals were recorded. The cramp frequency threshold was lower for the painful condition with respect to its baseline (mean ± SE, hypertonic saline: 25.7 ± 2.1 Hz, corresponding baseline: 31.2 ± 2.8 Hz; P < 0.01) while there was no difference between the threshold with isotonic injection with respect to baseline. EMG average rectified value and power spectral frequency were higher during the cramp than immediately before the stimulation that elicited the cramp (pre-cramp: 13.9 ± 1.6 μV and 75.4 ± 3.8 Hz, respectively; post-cramp: 19.9 ± 3.2 μV and 101.6 ± 6.0 Hz; P < 0.05). The results suggest that nociceptive muscle afferent activity induced by injection of hypertonic saline facilitates the generation of electrically elicited muscle cramps.     

Sodium balance during development of hypertension in the spontaneously hypertensive rat (SHR)

Sodium balance was studied in 7 and 16 week old male spontaneously hypertensive rats (SHR), in matched normotensive Wistar rats (NCR) and in Wistar Kyoto rats (WKR). The animals were placed in metabolic cages and given diets with either normal sodium content (5.35 mmol sodium/100 g food) or with a sodium content 3 or 10 times the normal. Whether on normal or increased sodium diet we did not observe any increased sodium retention in either SHR age group. However, in both SHR groups urinary sodium excretion was significantly decreased, while faecal sodium excretion was correspondingly increased compared with the controls. This shift of sodium excretion from kidneys to gastrointestinal tract in SHR did not reflect any ‘primary’ inability of the SHR kidneys to excrete sufficient sodium amounts since on high sodium diet they excreted the increased sodium load as readily as the normotensive controls. The present results do not support the concept that a primary renal retention of sodium and water should be of pathogenetic importance for the SHR variant of primary hypertension.     

Mechanism of acidic pH‐induced contraction in spontaneously hypertensive rat aorta: role of Ca2+ release from the sarcoplasmic reticulum

Aim: This study was conducted to investigate the mechanism of acidic pH‐induced contraction (APIC) with regard to Ca²⁺ handling using isometric tension recording experiments. Results: Decreasing extracellular pH from 7.4 to 6.5 produced a marked and sustained contraction of spontaneously hypertensive rat (SHR) aorta, that was 128.7 ± 2.0% of the 64.8 mm KCl‐induced contraction. Verapamil, an inhibitor of voltage‐dependent Ca²⁺ channels (VDCC) significantly inhibited the APIC. In Ca²⁺‐deficient solution, sustained contraction induced by acidic pH was abolished completely, while a transient contraction was still observed suggesting the release of Ca²⁺ from intracellular site. Ryanodine (1 μm ), a ryanodine receptor blocker, and 10 μm cyclopiazonic acid (CPA; a sarco/endoplasmic reticulum Ca²⁺ ATPase inhibitor) abolished the transient contraction induced by acidosis. In normal Ca²⁺‐containing solution, ryanodine significantly decreased the rate of rise as well as maximum level of APIC. Interestingly, ryanodine and CPA showed an additive inhibitory effect with verapamil and the combined treatment of ryanodine or CPA with verapamil nearly abolished the APIC. Conclusions: It is concluded that acidic pH induces Ca²⁺ release from ryanodine/CPA‐sensitive store of sarcoplasmic reticulum in SHR aorta. This Ca²⁺ plays an important role in the facilitation of the rate of rise of APIC, as well as contributing to the sustained contraction via a mechanism which is independent of Ca²⁺ influx through VDCC.     

依那普利对自发性高血压大鼠心肌ACE和ACE2表达的影响

 目的 观察自发性高血压大鼠(spontaneously hypertensive rats, SHR)心肌的血管紧张素转换酶(angiotensin-converting enzyme, ACE)和ACE2的表达,以及依那普利干预的影响。方法 将15只SHR随机分为2组：SHR对照组(n=7)和依那普利组(n=8),分别给以安慰剂、依那普利15mg.kg-1.d-1灌胃干预4周。干预结束后处死大鼠,分离左心室,行RT-PCR、western blot蛋白质免疫印迹检测。同步取10只WKY大鼠作为正常血压对照组。结果SHR心肌的ACE的mRNA和蛋白质的表达都显著高于)WKY组(1.68±0.34 vs 0.33±0.12, P＜0.05;1.21±0.14 vs 0.71±0.11, P＜0.05),而ACE2 的mRNA和蛋白质表达皆明显低于WKY组(0.50±0.15 vs 1.16±0.24, P＜0.05; 0.71±0.24 vs 1.22±0.14, P＜0.05)。依那普利明显降低ACE的mRNA和蛋白质表达(0.44±0.19 vs 1.68±0.34, P＜0.01; 0.87±0.13 vs 1.21±0.14, P＜0.05),提升ACE2的mRNA表达(1.77±0.49 vs 0.50±0.15, P＜0.05),对ACE2的蛋白表达无明显影响(0.42±0.22 vs 0.71±0.24, P＞0.05)。结论 SHR心肌ACE明显升高,ACE2显著降低,有利于血压上调。依那普利能降低ACE,提升ACE2,可能是血管紧张素转换酶抑制剂(angiotensin-converting enzyme inhibitors, ACEI)的降压机制之一。     

Immobilization stress induces vasodepressor and altered neuroendocrine responses in the adult stroke-prone spontaneously hypertensive male rat

The effects of acute (1 h) and daily repeated immobilization stress (14 days, twice-daily, 1 h) were studied on arterial blood pressure and heart rate and on the blood levels of several hormones in the adult (5 months old) stroke-prone spontaneously hypertensive rat (SHRSP) and in the age-matched normotensive Wistar-Kyoto (WKY) rat. The major result was the development of a long-lasting vasodepressor response in the SHRSP, while the same acute or repeated immobilization stress in the WKY rat led to the development of a prolonged vasopressor response. Differential changes to stress were also observed in practically all neuroendocrine axes with the exception of the pituitary-adrenal axis. The vasodepressor response to immobilization stress in SHRSP may be related to an exaggerated defence-like reaction causing an enhanced vasodilation in the skeletal muscle beds associated with a tachycardia similar to that in the normotensive control rats.     

Olmesartan restores the protective effect of remote ischemic perconditioning against myocardial ischemia/reperfusion injury in spontaneously hypertensive rats

OBJECTIVES:Remote ischemic perconditioning is the newest technique used to lessen ischemia/reperfusion injury. However, its effect in hypertensive animals has not been investigated. This study aimed to examine the effect of remote ischemic perconditioning in spontaneously hypertensive rats and determine whether chronic treatment with Olmesartan could influence the effect of remote ischemic perconditioning.METHODS:Sixty rats were randomly divided into six groups: vehicle-sham, vehicle-ischemia/reperfusion injury, vehicle-remote ischemic perconditioning, olmesartan-sham, olmesartan-ischemia/reperfusion and olmesartan-remote ischemic perconditioning. The left ventricular mass index, creatine kinase concentration, infarct size, arrhythmia scores, HIF–1α mRNA expression, miR-21 expression and miR-210 expression were measured.RESULTS:Olmesartan significantly reduced the left ventricular mass index, decreased the creatine kinase concentration, limited the infarct size and reduced the arrhythmia score. The infarct size, creatine kinase concentration and arrhythmia score during reperfusion were similar for the vehicle-ischemia/reperfusion group and vehicle-remote ischemic perconditioning group. However, these values were significantly decreased in the olmesartan-remote ischemic perconditioning group compared to the olmesartan-ischemia/reperfusion injury group. HIF–1α, miR-21 and miR-210 expression were markedly down-regulated in the Olmesartan-sham group compared to the vehicle-sham group and significantly up-regulated in the olmesartan-remote ischemic perconditioning group compared to the olmesartan-ischemia/reperfusion injury group.CONCLUSION:The results indicate that ¹ the protective effect of remote ischemic perconditioning is lost in vehicle-treated rats and that chronic treatment with Olmesartan restores the protective effect of remote ischemic perconditioning; ² chronic treatment with Olmesartan down-regulates HIF–1α, miR-21 and miR-210 expression and reduces hypertrophy, thereby limiting ischemia/reperfusion injury; and ³ recovery of the protective effect of remote ischemic perconditioning is related to the up-regulation of HIF–1α, miR-21 and miR-210 expression.     

心前区疼痛刺激对实验性大鼠急性心梗范围的影响

目的：观察心前区疼痛刺激对急性心梗大鼠心梗范围的影响。方法：SD雄性大鼠40只,随机分为4组：急性心梗对照组,心前区疼痛刺激组,六烃季(Hexamethonium,HEX)注射组,8-对苯磺酸基茶碱(8-p-sulfophenyl)theophylline,8-SPT)注射组。左心室连同室间隔横切,硝基四氮唑兰染色法测量心梗面积。结果：心前区疼痛刺激组较急性心梗对照组心梗面积显著减小。HEX注射组和8-SPT注射组与急性心梗对照组心梗面积无明显改变。结论：心前区疼痛刺激可能通过交感神经兴奋,间接起到心肌保护作用。     

Immune system of the spontaneously hypertensive rat: II. Morphology and function

The spontaneously hypertensive rat (SHR) is a stress-sensitive animal which exhibits moderate immune dysfunction that has been implicated in the onset of hypertension. In this study, we examined the morphology of SHR thymus and spleen and further characterized the immune deficiency using Wistar-Kyoto (WKY) and Fisher 344 (F-344) rats for comparison. The adult SHR thymus does not display the increase in medullary volume typically noted with aging and the volume density of the marginal zone is decreased in the spleen. In vivo tritiated-thymidine incorporation is also decreased in the spleen of unstimulated SHR. In mixed lymphocyte reactions (MLR), the proliferative response of SHR splenocytes is significantly decreased relative to controls, WKY and F-344. Addition of interleukin-1 (IL-1), interleukin-2 (IL-2), or indomethacin to the MLR cultures does not increase proliferation. The proliferative response to T cell receptor monoclonal antibody (mAb-TCR) or interleukin-2 (IL-2) are similarly impaired in the SHR. The depressed proliferative T cell response is reversed by prolactin. It is suggested that the SHR is a valuable model for the study of immune deficiency. © 1993 Wiley-Liss Inc.