Hypothermia In Rats Research Articles

Cold-Induced Thermogenesis and Inflammation-Associated Cold-Seeking Behavior Are Represented by Different Dorsomedial Hypothalamic Sites: A Three-Dimensional Functional Topography Study in Conscious Rats.

In the past, we showed that large electrolytic lesions of the dorsomedial hypothalamus (DMH) promoted hypothermia in cold-exposed restrained rats, but attenuated hypothermia in rats challenged with a high dose of bacterial lipopolysaccharide (LPS) in a thermogradient apparatus. The goal of this study was to identify the thermoeffector mechanisms and DMH representation of the two phenomena and thus to understand how the same lesion could produce two opposite effects on body temperature. We found that the permissive effect of large electrolytic DMH lesions on cold-induced hypothermia was due to suppressed thermogenesis. DMH-lesioned rats also could not develop fever autonomically: they did not increase thermogenesis in response to a low, pyrogenic dose of LPS (10 μg/kg, i.v.). In contrast, changes in thermogenesis were uninvolved in the attenuation of the hypothermic response to a high, shock-inducing dose of LPS (5000 μg/kg, i.v.); this attenuation was due to a blockade of cold-seeking behavior. To compile DMH maps for the autonomic cold defense and for the cold-seeking response to LPS, we studied rats with small thermal lesions in different parts of the DMH. Cold thermogenesis had the highest representation in the dorsal hypothalamic area. Cold seeking was represented by a site at the ventral border of the dorsomedial nucleus. Because LPS causes both fever and hypothermia, we originally thought that the DMH contained a single thermoregulatory site that worked as a fever–hypothermia switch. Instead, we have found two separate sites: one that drives thermogenesis and the other, previously unknown, that drives inflammation-associated cold seeking.SIGNIFICANCE STATEMENT Cold-seeking behavior is a life-saving response that occurs in severe systemic inflammation. We studied this behavior in rats with lesions in the dorsomedial hypothalamus (DMH) challenged with a shock-inducing dose of bacterial endotoxin. We built functional maps of the DMH and found the strongest representation of cold-seeking behavior at the ventral border of the dorsomedial nucleus. We also built maps for cold-induced thermogenesis in unanesthetized rats and found the dorsal hypothalamic area to be its main representation site. Our work identifies the neural substrate of cold-seeking behavior in systemic inflammation and expands the functional topography of the DMH, a structure that modulates autonomic, endocrine, and behavioral responses and is a potential therapeutic target in anxiety and panic disorders.

Clinical Vagus Nerve Stimulation Paradigms Induce Pronounced Brain and Body Hypothermia in Rats

Vagus nerve stimulation (VNS) is a widely used neuromodulation technique that is currently used or being investigated as therapy for a wide array of human diseases such as epilepsy, depression, Alzheimer's disease, tinnitus, inflammatory diseases, pain, heart failure and many others. Here, we report a pronounced decrease in brain and core temperature during VNS in freely moving rats. Two hours of rapid cycle VNS (7s on/18s off) decreased brain temperature by around [Formula: see text]C, while standard cycle VNS (30[Formula: see text]s on/300[Formula: see text]s off) was associated with a decrease of around [Formula: see text]C. Rectal temperature similarly decreased by more than [Formula: see text]C during rapid cycle VNS. The hypothermic effect triggered by VNS was further associated with a vasodilation response in the tail, which reflects an active heat release mechanism. Despite previous evidence indicating an important role of the locus coeruleus-noradrenergic system in therapeutic effects of VNS, lesioning this system with the noradrenergic neurotoxin DSP-4 did not attenuate the hypothermic effect. Since body and brain temperature affect most physiological processes, this finding is of substantial importance for interpretation of several previously published VNS studies and for the future direction of research in the field.

Effect of hypothermia on splenic leukocyte modulation and survival duration in severely septic rats

BackgroundTherapeutic hypothermia (HT) in severe septic shock is associated with prolonged survival. We hypothesized that moderate HT would prolong survival and modulate the inflammatory response in rats with septic shock by exerting its therapeutic effect on splenic leukocytes. Materials and methodsSevere septic shock was created in rats by cecal ligation and incision (CLI). One hour after CLI or laparotomy, rats were randomized to sham, normothermia (NT), or 4 h of HT followed by 2 h of rewarming. HT (31 ± 1°C) was induced using a cooling blanket and monitored via a rectal temperature probe. ResultsSurvival duration was 2.78 ± 1.0 h in NT rats and 8.33 ± 0.32 h in HT rats (n = 8/group, P < 0.0001). In separate groups, 3 h after CLI, the spleen weight was significantly smaller in NT rats (769 ± 100 mg) than in HT rats (947 ± 157 mg, P = 0.04). Fluorescent immunostaining of formyl peptide receptors on leukocytes in spleen tissue showed considerably higher formyl peptide receptor expression in HT rats than in NT rats. Significantly elevated proinflammatory cytokines and myeloperoxidase enzyme in plasma were found in NT rats compared with HT rats. Anti-inflammatory cytokine, interleukin-10, was significantly higher in HT rats. Both proinflammatory cytokines and plasma myeloperoxidase were significantly reduced in splenectomized NT rats. ConclusionsModerate hypothermic therapy significantly prolongs the survival duration of rats with severe septic shock. HT dampens the inflammatory response during septic shock by modulating the spleen to an anti-inflammatory mode and preventing the spleen from releasing activated splenic leukocytes into the blood.

Local Hypothermia as a Radioprotector of the Rectal Wall During Prostate Stereotactic Body Radiation Therapy

Tocompare the single-fraction dose-related incidence of rectal obstruction and/or bleeding in normothermic and hypothermic rectums of a rat model. A 1.9-cm length of rectum was irradiated with a single fraction in 57 Sprague-Dawley rats using a dedicated image-guided small animal irradiator and Monte Carlo-based treatment planning system. All rats had a rectal temperature control apparatus placed during irradiation and were stratified to achieve either a normothermic (37°C) or hypothermic (15°C) rectal wall temperature. Radiation was delivered to a 1-cm-diameter cylindrical volume about the cooling device and rectal wall. The radiation dose was escalated from 16Gy up to 37Gy to assess the dose response in each arm. The primary endpoint of this study was rectal obstruction and/or bleeding during a follow-up of 180 to 186days. Histologic scoring was performed on all study rats. Probit analysis showed a dose associated with a 50% incidence of rectal obstruction of 24.6Gy and 40.8Gy for normothermic and hypothermic arms, respectively. The occurrence of obstruction and/or bleeding correlated with the posttreatment histologic score for normothermic rats; however, there was no difference in histologic score between normothermic and hypothermic rats at the highest dose levels evaluated. A significant radioprotective effect was observed using local hypothermia during a single large dose of radiation for the functional endpoint of rectal obstruction and/or bleeding. A confirmatory study in a large animal model with anatomic and physiologic similarities to humans is suggested.

Influence of cooling medium on rat hemostatic system

The purpose of the article is to study the effect of air and water-immersion cooling media on the hemostatic system in the pre-active period of cold trauma. Material and methods. The study was performed on 43 laboratory Wistar rats. Single air hypothermia was modeled by placing animals in individual cells into a cooling chamber at a temperature –25 °С. The animals were in the chamber until the rectal temperature reached 30 °C, which corresponded to a moderate degree of hypothermia in rats. Water immersion hypothermia was modeled by placing animals in individual cells into water with a temperature 5 °C and air 7 °C. The criterion for cessation of exposure was a rectal temperature of 27–30 °C, which also corresponded to a moderate degree of hypothermia. We assessed the state of vascularplatelet and plasma hemostasis, as well as the physiological state of anticoagulant and fibrinolytic systems. The study was performed by using routine techniques and an integral method - thromboelastography. Results. It was established that during water immersion cooling, experimental animals developed thrombocytosis and activated their aggregation function. Laboratory indicators characterizing the initial stages of plasma hemostasis, and external and internal ways of activation did not change at this intensity of hypothermic exposure. At the same time, at the final stage of coagulation, pronounced thrombinemia was recorded, which was confirmed by a significant increase in the concentration of soluble fibrin-monomer complexes and a decrease in the time of their self-assembly. In addition, inhibition of fibrinolytic activity of plasma associated with a decrease in the concentration of antithrombin III was observed. In this case, single air hypothermia, accompanied by the rise in a rectal temperature up to 30 ° C, also caused significant changes in the hemostatic system. Vascular-platelet hemostasis responded to a significant decrease in aggregation activity of platelets. In plasma hemostasis, the final stage of coagulation was involved the most, which was manifested in hypocoagulant shifts. Along with this, pronounced inhibition of fibrinolytic activity of blood plasma was recorded. Thus, the described hemostasiological pictures indicate a pronounced influence of the environment causing supercooling on the response of an organism with hypothermia. In case of immersion hypothermia an increase in a rectal temperature up to 30 oС is accompanied by a pronounced activation of the coagulation processes and the thrombotic state of readiness. When supercooling with air after reaching a rectal temperature of 30 ° C, secondary hypocoagulation shifts are recorded.

Efficacy of ulinastatin for improvement of cerebral resuscitation using therapeutic hypothermia in rats with cardiac arrest

Objective To investigate the efficacy of ulinastatin for improvement of cerebral resuscitation using therapeutic hypothermia(TH)in rats with cardiac arrest(CA). Methods Sixty pathogen-free healthy male Sprague-Dawley rats, weighing 280-320 g, were divided into 5 groups(n=12 each)using a random number table: sham operation group(group Sham); CA/cardiopulmonary resuscitation(CPR)group(group CA/CPR); group TH; ulinastatin group(group U); ulinastatin + TH group(group U+ TH). CA was induced through transesophageal cardiac pacing, and CPR was performed at 4 min of untreated arrest to establish CA/CPR model.In group TH, the rectal temperature was cooled down to 32-34 ℃ by rubbing the body surface with ethanol within 15 min starting from the time point immediately after restoration of spontaneous circulation(ROSC), and maintained at this level for 6 h. In group U, ulinastatin 100 000 U/kg was injected via the femoral vein immediately after ROSC.In group U+ TH, ulinastatin 100 000 U/kg was intravenously injected immediately after ROSC, and rectal cooling was then started.The neurological function was evaluated and scored at 24 and 72 h after ROSC.The rats were sacrificed after the neurological function was scored at 72 h after ROSC, and the hippocampi were removed for microscopic examination of pathological changes in hippocampal CA1 region and for determination of caspase-3 expression(by immunohistochemical staining and Western blot)and Bax and Bcl-2 expression(by Western blot)in hippocampal tissues.Bax/Bcl-2 ratio was calculated. Results Compared with group Sham, the neurological function score was significantly decreased, the expression of caspase-3 and Bax was up-regulated, the expression of Bcl-2 was down-regulated, and Bax/Bcl-2 ratio was increased after ROSC in group CA/CPR(P<0.05). Compared with group CA/CPR, the neurological function score was significantly increased at 24 and 72 h after ROSC in TH and U+ TH groups and at 24 h after ROSC in group U, caspase-3 expression was significantly down-regulated, Bcl-2 expression was up-regulated, and Bax/Bcl-2 ratio was decreased at 72 h after ROSC in TH, U and U+ TH groups, and the expression of Bax was significantly down-regulated in U and U+ TH groups(P<0.05). Compared with group TH or group U, the neurological function score was significantly increased at 24 h after ROSC, and the expression of caspase-3 was down-regulated and Bax/Bcl-2 ratio was decreased at 72 h after ROSC in group U+ TH(P<0.05). The pathological changes of hippocampal tissues were significantly mitigated in group U+ TH when compared with group TH or group U. Conclusion Ulinastatin can improve the efficacy of TH for cerebral resuscitation in rats with cardiac arrest, and the mechanism may be related to inhibition of neuronal apoptosis. Key words: Trypsin inhibitors; Hypothermia, induced; Heart arrest; Cardiopulmonary resuscitation; Brain

Antidepressant & anxiolytic activities of N-(pyridin-3-yl) quinoxalin-2-carboxamide: A novel serotonin type 3 receptor antagonist in behavioural animal models.

Background & objectives:Alteration in the serotonin leads to the psychological illness, such as depression, anxiety, schizophrenia, eating disorders, obsessive-compulsive disorder, panic disorders and migraines. The objective of the current study was to investigate the antidepressant and anxiolytic activities of N-(pyridin-3-yl) quinoxalin-2-carboxamide (QCF-21), a novel 5-HT3 receptor antagonist in preclinical models of depression and anxiety.Methods:Antidepressant activity was evaluated in preliminary tests such as forced swim and tail suspension tests (FST & TST). Anti-anxiety effect of QCF-21 was investigated by employing elevated plus maze (EPM), light/dark and hole board tests. Olfactory bulbectomy (OBX) in rats was used as chronic model of depression. Mechanistic test of QCF-21 was evaluated by reserpine-induced hypothermia and 5-hydroxytryptophan (5-HTP)-induced head-twitch response.Results:The dose-response study revealed an initial antidepressant-like effect of QCF-21(0.25-1 mg/kg, i.p.) in the FST and TST and anxiolytic-like effect in EPM, light and dark and hole board tests. QCF-21 potentiated the 5-HTP-induced head-twitches response in mice and reversed reserpine-induced hypothermia in rats. QCF-21 significantly reversed the behavioural anomalies post-OBX in rats.Interpretation & conclusions:The present findings indicate the potential antidepressant-like and anxiolytic-like effects of QCF-21 at low doses in rodent behavioural models of depression and anxiety. Further studies need to be done to understand the underlying mechanism.

Novel method for inducing rapid, controllable therapeutic hypothermia in rats using a perivascular implanted closed-loop cooling circuit

BackgroundHypothermia is the most potent protective therapy available for cerebral ischemia. In experimental models, cooling the brain even a single degree Celsius alters outcome after global and focal ischemia. Difficulties translating therapeutic hypothermia to patients with stroke or after cardiac arrest include: uncertainty as to the optimal treatment duration; best target-depth temperature; and longest time delay after which therapeutic hypothermia won’t benefit. Recent results from human clinical trials suggest that cooling with surface methods provides insufficient cooling speed or control over target temperature. Comparison with existing methodsAvailable animal models incorporate surface cooling methods that are slow, and do not allow for precise control of the target temperature. New methodTo address this need, we developed a rapid, simple, inexpensive model for inducing hypothermia using a perivascular implanted closed-loop cooling circuit. The method allows precise control of the target temperature. ResultsUsing this method, target temperature for therapeutic hypothermia was reached within 13±1.07min (Mean±SE). Once at target, the temperature was maintained within 0.09°C for 4h. ConclusionsThis method will allow future experiments to determine under what conditions therapeutic hypothermia is effective, determine the optimal relationship among delay, duration, and depth, and provide the research community with a new model for conducting further research into mechanistic questions underlying the efficacy of therapeutic hypothermia.

Artificial hypothermia in rats, unlike natural hibernation in ground squirrels Spermophilus undulatus, is not accompanied by the inhibition of respiration in liver mitochondria

There are different assumptions regarding mechanisms of the suppression of oxidative phosphorylation in liver mitochondria of hibernating animals. In this work, the data suggesting that inhibition of respiratory chain in the segment of complex III may be the main cause of this suppression are presented. A significant inhibition of phosphorylation and maximum rates of respiration in hibernating animals as compared to active animals was demonstrated for liver mitochondria of ground squirrels Spermophilus undulatus using substrates oxidized through complex I (glutamate, pyruvate), as well as through complex II (succinate), while no changes in functioning of complex IV upon oxidation of artificial substrate TMPD oxidized via cytochrome c were detected. This indicates that the point of inhibition of mitochondrial respiration is localized in the area between complex II and cytochrome c. It is unclear whether such inhibition is necessary for the induction of natural hibernation state or it is the consequence of any hypothermia. We used homeothermic animals (rats), which were brought into a state of nondrug hypometabolism under conditions of hypoxia, hypercapnia and low ambient temperature, leading to a decrease of body temperature down to 15°C, and ground squirrels having the same temperature during the entrance into hibernation. Artificial hypometabolism caused by hypothermia in non-hibernating mammals was not accompanied by the inhibition of respiration in liver mitochondria, which is specific to the natural hypometabolic state in hibernating ground squirrels.

Cerebral Blood Flow Threshold Is Higher for Membrane Repolarization Than for Depolarization and Is Lowered by Intraischemic Hypothermia in Rats.

To evaluate the cerebral blood flow thresholds for membrane depolarization and repolarization and the effect of brain hypothermia on the cerebral blood flow threshold for membrane repolarization. Prospective animal study. Experimental laboratory in a university hospital. Male Sprague-Dawley rats (n = 40). Cerebral blood flow and membrane depolarization and repolarization in the cerebral cortex were simultaneously monitored by laser Doppler and extracellular potential, respectively. Following bilateral occlusion of the common carotid arteries, cerebral blood flow was decreased by draining blood at a rate of 2.5% of the control level/min until membrane depolarization was initiated. At 5 and 10 minutes (Normothermia 5 and Normothermia 10 groups, respectively) after depolarization onset, cerebral blood flow was restored at the same rate until membrane repolarization was observed. In some animals, intraischemic brain hypothermia targeting 31°C was initiated immediately after the onset of depolarization (Hypothermia 5 and Hypothermia 10 groups). The cerebral blood flow threshold for repolarization (46.5% ± 12%) was significantly higher than that for depolarization (18.9% ± 4.8%; p < 0.01) in the Normothermia 5 group and was further increased to 61.5% ± 14% (p < 0.01) in the Normothermia 10 group. With initiation of hypothermia, the cerebral blood flow threshold for membrane repolarization was suppressed to 33.8% ± 10% in the Hypothermia 5 group (p < 0.01 vs Normothermia 5 group) and was unaltered by prolongation of ischemia (Hypothermia 10 group; 36.6% ± 6%). Cerebral blood flow thresholds were significantly higher for repolarization than for depolarization and were further increased by prolonged ischemia. Intraischemic brain hypothermia decreased the repolarization threshold and abrogated the increase in the repolarization threshold caused by prolonged ischemia.

Restoration of Vital Activity in Deeply Cooled Animals Using Physiological Methods without Rewarming

White male Wistar rats were cooled in water (9–10°C) to respiratory arrest (rectal temperature 14.7 ± 0.5°C, brain temperature 16.0 ± 0.3°C). After removal from water, animals received intravenous 0.5% ethylenediaminetetraacetic acid disodium salt (Na2EDTA) to decrease the blood Ca2+ concentration. Respiration was reinitiated 4–8 min after administration of Na2EDTA. One group of rats was then further cooled to a body temperature of 12.2 ± 0.7°C (brain temperature 14.9 ± 0.3°C); at these low temperatures, rats maintained respiration at a level of 12 ± 3 cycles/min for 2–3 h. After restarting respiration, rats of group 2 were not subjected to further cooling, and the same dose of Na2EDTA, at rectal and brain temperatures of 15–16°C, increased respiratory rate to the higher level of 29 ± 4 cycles/min. Administration of Na2EDTA into the circulatory system of hypothermic rats evidently slowed the process of accumulation of Ca2+ in the cytoplasm of respiratory center cells, impairing their functioning. These results suggest that in hypothermia, injection of Na2EDTA activates the mechanisms protecting cells from cold injury. These results, along with published data, lead to the conclusion that non-hibernating homeothermic organisms are able to maintain the main vital functions at signifi cantly reduced body temperatures.

Platelet-activating factor is a potent pyrogen and cryogen, but it does not mediate lipopolysaccharide fever or hypothermia

We examined whether platelet-activating factor (PAF) and its receptor mediate lipopolysaccharide (LPS)-induced fever and hypothermia in rats. Two highly potent, structurally distinct antagonists of the PAF receptor, CV6209 and WEB2086, were used. At a neutral ambient temperature (Ta) of 30ºC, administration of LPS at a low (10 μg/kg, i.v.) or high (1,000 μg/kg, i.v.) dose resulted in fever. The response to the high dose was turned into hypothermia at a subneutral Ta of 22ºC. Neither LPS-induced fever nor hypothermia was affected by pretreatment with CV6209 (5 mg/kg, i.v.) or WEB2086 (5 mg/kg, i.v.). However, both PAF antagonists were efficacious in blocking the thermoregulatory response caused by PAF (334 pmol/kg/min, 1 h, i.v.), regardless of whether the response was a fever (at 30ºC) or hypothermia (at 22ºC). Additional experiments showed that the thermoregulatory responses to LPS and PAF are also distinct in terms of their mediation by prostaglandins. Neither PAF fever nor PAF hypothermia was affected by pretreatment with the cyclooxygenase-2 inhibitor SC236 (5 mg/kg, i.p.), which is known to abrogate LPS fever. The responses to PAF were also unaffected by pretreatment with the cyclooxygenase-1 inhibitor SC560 (5 mg/kg, i.p.), which is known to attenuate LPS hypothermia. In conclusion, PAF infusion at a picomolar dose causes fever at thermoneutrality but hypothermia in a subthermoneutral environment, both responses being dependent on the PAF receptor and independent of prostaglandins. However, the PAF receptor does not mediate LPS-induced fever or hypothermia, thus challenging the dogma that PAF is an upstream mediator of responses to LPS.

Moderate Hypothermia Significantly Decreases Hippocampal Cell Death Involving Autophagy Pathway after Moderate Traumatic Brain Injury.

Here, we evaluated changes in autophagy after post-traumatic brain injury (TBI) followed by moderate hypothermia in rats. Adult male Sprague-Dawley rats were randomly divided into four groups: sham injury with normothermia group (37 °C); sham injury with hypothermia group (32 °C); TBI with normothermia group (TNG; 37 °C); and TBI with hypothermia group (THG; 32 °C). Injury was induced by a fluid percussion TBI device. Moderate hypothermia (32 °C) was achieved by partial immersion in a water bath (0 °C) under general anesthesia for 4 h. All rats were killed at 24 h after fluid percussion TBI. The ipsilateral hippocampus in all rats was analyzed with hematoxylin and eosin staining; terminal deoxynucleoitidyl transferase-mediated nick end labeling staining was used to determine cell death in ipsilateral hippocampus. Immunohistochemistry and western blotting of microtubule-associated protein light chain 3 (LC3), Beclin-1, as well as transmission electron microscopy performed to assess changes in autophagy. At 24 h after TBI, the cell death index was 27.90 ± 2.36% in TNG and 14.90 ± 1.52% in THG. Expression level of LC3 and Beclin-1 were significantly increased after TBI and were further up-regulated after post-TBI hypothermia. Further, ultrastructural observations showed that there was a marked increase of autophagosomes and autolysosomes in ipsilateral hippocampus after post-TBI hypothermia. Our data demonstrated that moderate hypothermia significantly attenuated cell death and increased autophagy in ipsilateral hippocampus after fluid percussion TBI. In conclusion, autophagy pathway may participate in the neuroprotective effect of post-TBI hypothermia.

5'-adenosine monophosphate-induced hypothermia attenuates brain ischemia/reperfusion injury in a rat model by inhibiting the inflammatory response.

Hypothermia treatment is a promising therapeutic strategy for brain injury. We previously demonstrated that 5′-adenosine monophosphate (5′-AMP), a ribonucleic acid nucleotide, produces reversible deep hypothermia in rats when the ambient temperature is appropriately controlled. Thus, we hypothesized that 5′-AMP-induced hypothermia (AIH) may attenuate brain ischemia/reperfusion injury. Transient cerebral ischemia was induced by using the middle cerebral artery occlusion (MCAO) model in rats. Rats that underwent AIH treatment exhibited a significant reduction in neutrophil elastase infiltration into neuronal cells and matrix metalloproteinase 9 (MMP-9), interleukin-1 receptor (IL-1R), tumor necrosis factor receptor (TNFR), and Toll-like receptor (TLR) protein expression in the infarcted area compared to euthermic controls. AIH treatment also decreased the number of terminal deoxynucleotidyl transferase dUTP nick end labeling- (TUNEL-) positive neuronal cells. The overall infarct volume was significantly smaller in AIH-treated rats, and neurological function was improved. By contrast, rats with ischemic brain injury that were administered 5′-AMP without inducing hypothermia had ischemia/reperfusion injuries similar to those in euthermic controls. Thus, the neuroprotective effects of AIH were primarily related to hypothermia.

Drug-induced Hypothermia by 5HT1A Agonists Provide Neuroprotection in Experimental Stroke: New Perspectives for Acute Patient Treatment

Drug-induced hypothermia reduces brain damage in animal stroke models and is an undiscovered potential in human stroke treatment. We studied hypothermia induced by the serotonergic agonists S14671 (1-[2-(2-thenoylamino)ethyl]-4[1-(7- methoxynaphtyl)]piperazine) and ipsapirone in a rat stroke model and in man by literature meta-analysis. Rats had 60 minutes of middle cerebral artery occlusion (MCAO) and then 7 days of survival. Body temperatures were monitored for 22 hours. Thirty minutes after MCAO, 1 group (n = 9) received bolus of S14671 (.75 mg/kg) and continuous infusion of .06 mg/kg hour(-1) S14671 for 20 hours. Other MCAO rats (n = 7) had bolus of ipsapirone (.75 mg/kg) and continuous infusion of .25 mg/kg hour(-1) ipsapirone for 3 hours. Controls (n = 9; n = 5) received similar amounts of vehicle as bolus and continuous infusion for 20 hours/3 hours. Additional controls of the S14761 effect in MCAO were performed as previously mentioned (n = 10) but with rats kept normothermic by a heating lamp for 22 hours. Finally, a meta-analysis of ipsapirone-induced hypothermia in man was included. Infarct volumes were reduced by 50% in hypothermic rats versus controls (P < .05). S14671 rats kept normothermic did not show infarct reduction (P > .05). The body temperature after stroke was reduced 1.0-3.0°C compared with controls for 20 hours with S14671 treatment and for 6 hours with ipsapirone treatment. In humans, ipsapirone reduced temperature in average with .55 °C ranging between .1-1.4 °C. 5-hydroxytryptamine receptor 1A (5HT(1A)) agonists significantly reduce infarct volumes in MCAO rats primarily because of the hypothermic drug effect. 5HT(1A) agonists may be introduced to reduce body temperatures rapidly and prepare patients for further therapeutic hypothermia.

Cardiovascular effects of levosimendan during rewarming from hypothermia in rat

BackgroundPrevious research aimed at ameliorating hypothermia-induced cardiac dysfunction has shown that inotropic drugs, that stimulate the cAMP, – PKA pathway via the sarcolemmal β-receptor, have a decreased inotropic effect during hypothermia. We therefore wanted to test whether levosimendan, a calcium sensitizer and dose-dependent phosphodiesterase 3 (PDE3) inhibitor, is able to elevate stroke volume during rewarming from experimental hypothermia. MethodsA rat model designed for circulatory studies during experimental hypothermia (4h at 15°C) and rewarming was used. The following three groups were included: (1) A normothermic group receiving levosimendan, (2) a hypothermic group receiving levosimendan the last hour of stable hypothermia and during rewarming, and (3) a hypothermic placebo control group. Hemodynamic variables were monitored using a Millar conductance catheter in the left ventricle (LV), and a pressure transducer connected to the left femoral artery. In order to investigate the level of PKA stimulation by PDE3 inhibition, myocardial Ser23/24-cTnI phosphorylation was measured using Western-blot. ResultsAfter rewarming, stroke volume (SV), cardiac output (CO) and preload recruitable stroke work (PRSW) were restored to within pre-hypothermic values in the levosimendan-treated animals. Compared to the placebo group after rewarming, SV, CO, PRSW, as well as levels of Ser23/24-cTnI phosphorylation, were significantly higher in the levosimendan-treated animals. ConclusionThe present data shows that levosimendan ameliorates hypothermia-induced systolic dysfunction by elevating SV during rewarming from 15°C. Inotropic treatment during rewarming from hypothermia in the present rat model is therefore better achieved through calcium sensitizing and PDE3 inhibition, than β-receptor stimulation.

The effect of controlled mild hypothermia on large scald burns in a resuscitated rat model.

ObjectiveEarly surface cooling of burns reduces pain, depth of injury and improves healing. We hypothesized that controlled mild hypothermia would also prolong survival in a fluid resuscitated rat model of large scald burns.MethodsForty rats were anesthetized and a single full-thickness scald burn covering 40% of total body surface area was created on each of the rats. The rats were then randomized to hypothermia (n=20) or no hypothermia (n=20). Mild hypothermia (a reduction of 2°C) was induced with intraperitoneal 4°C normal saline and ice packs. After 2 hours of hypothermia, the rats were rewarmed back to their baseline temperature with a heating pad. The control rats received room temperature intraperitoneal saline. The difference in survival between the groups was determined using Kaplan-Meier analysis and the log-rank test.ResultsHypothermia was induced in all experimental rats within a mean of 22 minutes (95% confidence interval, 17 to 27). The number of normothermic and hypothermic rats that expired at each time interval were: at 1 hour, 4 vs. 0; at 10 hours, 2 from each group; at 24 hours, 0 vs. 1; at 48 hours, 2 vs. 2; at 72 hours, 1 vs. 1; and at 120 hours, 1 vs. 1 respectively. There were no differences in time to survival between the groups.ConclusionInduction of brief, mild hypothermia does not prolong survival in a resuscitated rat model of large scald burns.

Changes in cardiac thrombomodulin and heat shock transcription factor 1 expression and peripheral thrombomodulin and catecholamines during hypothermia in rats

Effects of hypothermia and rewarming on thrombomodulin, catecholamines and heat shock transcription factor 1 (HSF1) were studied in rats. The aims of this study were to clarify whether cold stress, under anesthesia, is sufficient to change levels of thrombomodulin in healthy endothelium and in the circulation and whether adrenaline, noradrenaline and HSF1 could act as regulators in the process. Rats were divided into control, mild hypothermia (2 and 4.5 hours at + 21 °C; MH1, MH2), severe hypothermia (2 and 4.5 h at + 10 °C; SH1, SH2) and two rewarming groups (2 h at + 10 °C followed by 2 h at + 21 °C or 3 h at + 28 °C; SHW1, SHW2) (n = 15/group, except n = 6 in MH1). Fentanyl-fluanisone-midazolam was used as anesthetic. Low levels of thrombomodulin in plasma and myocardial arterioles/venules measured by ELISA and immunohistochemistry were associated with significant increase of thrombomodulin transcript level in SH1 rats analyzed by quantitative PCR. Plasma adrenaline correlated negatively with the relative amount of myocardial thrombomodulin transcripts and positively with plasma thrombomodulin in SH. Transcript levels of thrombomodulin and HSF1 correlated strongly (r = 0.83; p < 0.001) in SH. Plasma/urine ratio of thrombomodulin and plasma adrenaline (r = 0.87; p = 0.005) or noradrenaline (r = 0.78; p = 0.023) were strongly correlated in SHW1 rats. Hence, cellular and soluble levels of thrombomodulin are modified by cold stress in healthy rats, possibly via catecholamines and HSF1.

Neuroprotective effect of epidural hypothermia after spinal cord lesion in rats

OBJECTIVES: To evaluate the neuroprotective effect of epidural hypothermia in rats subjected to experimental spinal cord lesion.METHODS:Wistar rats (n = 30) weighing 320-360 g were randomized to two groups (hypothermia and control) of 15 rats per group. A spinal cord lesion was induced by the standardized drop of a 10-g weight from a height of 2.5 cm, using the New York University Impactor, after laminectomy at the T9-10 level. Rats in the hypothermia group underwent epidural hypothermia for 20 minutes immediately after spinal cord injury. Motor function was assessed for six weeks using the Basso, Beattie and Bresnahan motor scores and the inclined plane test. At the end of the final week, the rats' neurological status was monitored by the motor evoked potential test and the results for the two groups were compared.RESULTS:Analysis of the Basso, Beattie and Bresnahan scores obtained during the six-week period indicated that there were no significant differences between the two groups. There was no significant difference between the groups in the inclined plane test scores during the six-week period. Furthermore, at the end of the study, the latency and amplitude values of the motor evoked potential test were not significantly different between the two groups.CONCLUSION:Hypothermia did not produce a neuroprotective effect when applied at the injury level and in the epidural space immediately after induction of a spinal cord contusion in Wistar rats.

Fuzi-Lizhong pill compensates hypothyroid-hypothermia via ghrelin release

Ethnopharmacological relevanceFuzi-Lizhong pill (FLZ) is a traditional Chinese medicine for treating patients with Spleen Yang deficient syndrome. Ghrelin, a peptide with 28 amino acid residues, plays multiple roles in thermogenesis. This study aims to explore FLZ regulating ghrelin to compensate hypothermia in rats with hypothyroid and indigestion. Materials and methodsIn litter-matched rats, hypothermia was developed with both thyroidectomy at d1 and interscapular brown adipose (IBA) removal at d42, indigestion was induced with both high fat diet and fasting-feeding cycle from d56; the littermates with hypothermia and indigestion were administrated with FLZ from d70. Adaptive thermogenesis, thyroid hormones, metabolites, ghrelin dynamics were measured at d98. ResultsThe results showed that plasma ghrelin levels were inversely correlated with the gastric ghrelin levels and adaptive thermogenesis in rats undergone both thyroidectomy and IBA removal. Fatty diet and FLZ enhanced the increase of plasma ghrelin of hypothyroid rats. These were supported by the changes of plasma thyroid related hormones, plasma metabolites, gastric ghrelin mRNA and protein, and the effects of fatty diet or FLZ. ConclusionsOur results suggest that more ghrelin release compensate chronic hypothermia in rats with both hypothyroidism and indigestion. It could explain the mechanisms of FLZ in relieving chronic hypothermia.