RESUMO
BACKGROUND: Brain death (BD) in potential organ donors is responsible for hemodynamic instability and organ hypoperfusion, leading to myocardial dysfunction. Hypertonic saline (HS) is a volume expander with positive effects on hemodynamics and immunomodulation and was tested in this study to prevent left ventricular (LV) dysfunction and myocardial injury. METHODS: BD was induced in anesthetized Wistar rats by inflating a subdural balloon catheter, except in sham-operated animals (n = 6). After BD induction, Control animals received only normal saline solution (NaCl 0.9%, 4 mL/kg; n = 6), and treated animals were divided to receive HS (NaCl, 7.5% 4 mL/kg) at 1 min (HS1, n = 6) or 60 min (HS60, n = 6) thereafter. We continuously assessed cardiac function for 6 h with LV pressure-volume analysis. Inflammatory response, markers of myocardial injury, and cellular apoptosis-related proteins were investigated. RESULTS: BD was associated with decreased LV systolic and diastolic function. In comparison with the Control group, HS treatments improved LV ejection fraction (HS1, 51% [40-66]; HS60, 71% [28-82]; Control, 46% [23-55]; P < 0.05) and other parameters of LV systolic function 6 h after BD induction. However, no ventricular relaxation advantages were observed during the same period. HS treatments increased antiapoptotic protein expression and decreased vascular adhesion molecule and tumor necrosis factor alpha expression. No significant differences in histologic or structural protein changes were observed between groups. CONCLUSIONS: The observed data suggest that HS ameliorates LV systolic dysfunction and seems to reduce myocardial tissue compromise in BD rats, even when the treatment is performed during the process triggered by this event.
Assuntos
Morte Encefálica/fisiopatologia , Miocárdio/patologia , Solução Salina Hipertônica/uso terapêutico , Disfunção Ventricular Esquerda/prevenção & controle , Animais , Morte Encefálica/patologia , Hemodinâmica/efeitos dos fármacos , Masculino , Ratos , Ratos Wistar , Sódio/sangueRESUMO
BACKGROUND: Experimental hemorrhagic shock (HS) is based on controlling bleeding and the treatment of fluid resuscitation to restore tissue oxygenation and perfusion. The HS could promote ischemia/reperfusion injury, which induces a general exacerbation of the inflammatory process, initially compromising the lungs. N-acetylcysteine (NAC), an antioxidant, may attenuate ischemia/reperfusion injury. This study evaluated the effect of NAC in association with fluid resuscitation on pulmonary injury in a controlled HS model in rats. METHODS: Male Wistar rats were submitted to controlled HS (mean arterial pressure of 35 mm Hg for 60 min). Two groups were constituted according to resuscitation solution administered: RLG (Ringer's lactate solution) and RLG+NAC (Ringer's lactate in association with 150 mg/kg NAC. A control group was submitted to catheterization only. After 120 min of resuscitation, bronchoalveolar lavage was performed to assess intra-alveolar cell infiltration and pulmonary tissue was collected for assessment of malondialdehyde, interleukin 6, and interleukin 10 and histopathology. RESULTS: Compared with the RLG group, the RLG+NAC group showed lower bronchoalveolar lavage inflammatory cell numbers, lower interstitial inflammatory infiltration in pulmonary parenchyma, and lower malondialdehyde concentration. However, tissue cytokine (interleukin 6 and interleukin 10) expression levels were similar. CONCLUSION: N-acetylcysteine was associated with fluid resuscitation-attenuated oxidative stress and inflammatory cell infiltration in pulmonary parenchyma. N-acetylcysteine did not modify cytokine expression.
Assuntos
Acetilcisteína/farmacologia , Acetilcisteína/uso terapêutico , Lesão Pulmonar Aguda/prevenção & controle , Pulmão/efeitos dos fármacos , Pulmão/patologia , Traumatismo por Reperfusão/prevenção & controle , Choque Hemorrágico/complicações , Lesão Pulmonar Aguda/metabolismo , Lesão Pulmonar Aguda/patologia , Animais , Antioxidantes/farmacologia , Antioxidantes/uso terapêutico , Modelos Animais de Doenças , Hidratação , Interleucina-10/metabolismo , Interleucina-6/metabolismo , Pulmão/metabolismo , Masculino , Malondialdeído/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Ratos , Ratos Wistar , Traumatismo por Reperfusão/metabolismo , Traumatismo por Reperfusão/patologia , RessuscitaçãoRESUMO
OBJECTIVES: Pulmonary arterial hypertension (PAH) is a cardiopulmonary disease that affects the pulmonary vasculature, leading to increased afterload and eventually right ventricular (RV) remodelling and failure. Bilateral sympathectomy (BS) has shown promising results in dampening cardiac remodelling and dysfunction in several heart failure models. In the present study, we investigated whether BS reduces pulmonary arterial remodelling and mitigates RV remodelling and failure. METHODS: PAH was induced in male Wistar rats by intraperitoneal injection of monocrotaline. Rats were divided into 3 groups, involving untreated PAH (n = 15), BS-treated PAH (n = 13) and non-manipulated control rats (n = 13). Three weeks after PAH induction, the rats were anaesthetized and RV function was assessed via the pressure-volume loop catheter approach. Upon completion of the experiment, the lungs and heart were harvested for further analyses. RESULTS: BS was found to prevent pulmonary artery remodelling, with a clear reduction in α-smooth muscle actin and endothelin-1 expression. RV end-systolic pressure was reduced in the BS group, and preload recruitable stroke work was preserved. BS, therefore, mitigated RV remodelling and cardiomyocyte hypertrophy and diminished oxidative stress. CONCLUSIONS: We showed that thoracic BS may be an important treatment option for PAH patients. Blockade of the sympathetic pathway can prevent pulmonary remodelling and protect the RV from oxidative stress, myocardial remodelling and function decay.
Assuntos
Hipertensão Pulmonar , Hipertensão Arterial Pulmonar , Disfunção Ventricular Direita , Animais , Modelos Animais de Doenças , Hipertensão Pulmonar Primária Familiar , Humanos , Masculino , Estresse Oxidativo , Artéria Pulmonar , Ratos , Ratos Wistar , Simpatectomia , Remodelação Vascular , Função Ventricular Direita , Remodelação VentricularRESUMO
BACKGROUND: Brain death (BD) is associated with systemic inflammatory compromise, which might affect the quality of the transplanted organs. This study investigated the expression profile of cardiac microRNAs (miRNAs) after BD, and their relationship with the observed decline in myocardial function and with the changes induced by hypertonic saline solution (HSS) treatment. METHODS: Wistar rats were assigned to sham-operation (SHAM) or submitted to BD with and without the administration of HSS. Cardiac function was assessed for 6 h with left ventricular (LV) pressure-volume analysis. We screened 641 rodent miRNAs to identify differentially expressed miRNAs in the heart, and computational and functional analyses were performed to compare the differentially expressed miRNAs and find their putative targets and their related enriched canonical pathways. RESULTS: An enhanced expression in canonical pathways related to inflammation and myocardial apoptosis was observed in BD induced group, with 2 miRNAs, miR-30a-3p, and miR-467f, correlating with the level of LV dysfunction observed after BD. Conversely, HSS treated after BD and SHAM groups showed similar enriched pathways related to the maintenance of heart homeostasis regulation, in agreement with the observation that both groups did not have significant changes in LV function. CONCLUSIONS: These findings highlight the potential of miRNAs as biomarkers for assessing damage in BD donor hearts and to monitor the changes induced by therapeutic measures like HSS, opening a perspective to improve graft quality and to better understand the pathophysiology of BD. The possible relation of BD-induced miRNA's on early and late cardiac allograft function must be investigated.
Assuntos
Transplante de Coração , MicroRNAs , Animais , Morte Encefálica , Transplante de Coração/efeitos adversos , Humanos , MicroRNAs/genética , Ratos , Ratos Wistar , Solução Salina Hipertônica/farmacologia , Solução Salina Hipertônica/uso terapêutico , Doadores de TecidosRESUMO
Surgical neuromodulation therapies are still considered a last resort when standard therapies have failed for patients with progressive heart failure (HF). Although a number of experimental studies have provided robust evidence of its effectiveness, the lack of strong clinical evidence discourages practitioners. Thoracic unilateral sympathectomy has been extensively studied and has failed to show significant clinical improvement in HF patients. Most recently, bilateral sympathectomy effect was associated with a high degree of success in HF models, opening the perspective to be investigated in randomized controlled clinical trials. In addition, a series of clinical trials showed that bilateral sympathectomy was associated with a decreased risk of sudden death, which is an important outcome in patients with HF. These aspects indicates that bilateral sympathectomy could be an important alternative in the treatment of HF wherein pharmacological treatment barely reaches the target dose.
Assuntos
Insuficiência Cardíaca , Hiperidrose , Procedimentos Cirúrgicos Torácicos , Insuficiência Cardíaca/cirurgia , Humanos , Hiperidrose/cirurgia , Simpatectomia , Resultado do TratamentoRESUMO
INTRODUCTION: Supraceliac aortic clamping in major vascular procedures promotes splanchnic ischemia and reperfusion (I/R) injury that may induce endothelial dysfunction, widespread inflammation, multiorgan dysfunction, and death. We tested the hypothesis that local or remote ischemic preconditioning (IPC) may be protective against injury after supraceliac aortic clamping through the modulation of mesenteric leukocyte-endothelial interactions, as evaluated with intravital microscopy and expression of adhesion molecules. METHODS: Fifty-six male Wistar rats (weight, 190 to 250 g), were divided into four groups of 14 rats each: control-sham surgery without aortic occlusion; I/R through supraceliac aortic occlusion for 20 minutes, followed by 120 minutes of reperfusion; local IPC through supraceliac aortic occlusion for two cycles of 5 minutes of ischemia and 5 minutes of reperfusion, followed by the same protocol of the IR group; remote IPC through infrarenal aortic occlusion for two cycles of 10 minutes of ischemia and 10 minutes of reperfusion, followed by the same protocol of the IR group. Seven animals per group were used to evaluate in vivo leukocyte-endothelial interactions in postcapillary venules with intravital microscopy and another seven animals per group were used to collect mesentery samples for immunohistochemistry demonstration of adhesion molecules expression. RESULTS: Supraceliac aortic occlusion increased the number of rolling leukocytes with slower velocities and increased the number of adherent leukocytes to the venular surface and leukocyte migration to the interstitium. The expression of P-selectin, E-selectin, and intercellular adhesion molecule-1 was also increased significantly after I/R. Local or remote IPC reduced the leukocyte recruitment in vivo and normalized the expression of adhesion molecules. CONCLUSIONS: Local or remote IPC reduces endothelial dysfunction on mesenteric microcirculation caused by I/R injury after supraceliac aortic clamping.
Assuntos
Aorta/cirurgia , Moléculas de Adesão Celular/metabolismo , Células Endoteliais/imunologia , Precondicionamento Isquêmico/métodos , Migração e Rolagem de Leucócitos , Traumatismo por Reperfusão/prevenção & controle , Circulação Esplâncnica , Procedimentos Cirúrgicos Vasculares/efeitos adversos , Animais , Constrição , Selectina E/metabolismo , Imuno-Histoquímica , Molécula 1 de Adesão Intercelular/metabolismo , Leucócitos/imunologia , Masculino , Microcirculação , Microscopia de Vídeo , Selectina-P/metabolismo , Ratos , Ratos Wistar , Traumatismo por Reperfusão/etiologia , Traumatismo por Reperfusão/imunologia , Traumatismo por Reperfusão/fisiopatologia , Fatores de Tempo , Vênulas/imunologia , Vênulas/fisiopatologiaRESUMO
OBJECTIVE: Brain death (BD) triggers important hemodynamic and inflammatory alterations, compromising the viability of organs suitable for transplantation. To better understand the microcirculatory alterations in donor lungs caused by BD. The present study investigated the pulmonary microcirculation in a rodent model of BD via intravital microscopy. METHODS: Male Wistar rats were anaesthetized and mechanically ventilated. They were trepanned and BD was induced through the increase in intracranial pressure. As control group, sham-operated (SH) rats were trepanned only. In both groups, expiratory O2 and CO2 were monitored and after three hours, a thoracotomy was performed, and a window was created to observe the lung surface using an epi-fluorescence intravital microscopy. Lung expression of intercellular adhesion molecule (ICAM)-1 and endothelial nitric oxide synthase (eNOS) was evaluated by immunohistochemistry, and cytokines were measured in lung samples. RESULTS: Three hours after the surgical procedures, pulmonary perfusion was 73% in the SH group. On the other hand, BD animals showed an important decrease in organ perfusion to 28% (p = 0.036). Lung microcirculatory compromise after BD induction was associated with an augmentation of the number of leukocytes recruited to lung tissue, and with a reduction in eNOS expression and an increase in ICAM-1 expression on lung endothelial cells. BD rats showed higher values of expiratory O2 and lower values of CO2 in comparison with SH animals after three hours of monitoring. CONCLUSION: Data presented showed that BD triggers an important hypoperfusion and inflammation in the lungs, compromising the donor pulmonary microcirculation.
OBJETIVO: A morte cerebral (MC) desencadeia alterações hemodinâmicas e inflamatórias importantes, comprometendo a viabilidade dos órgãos empregados em transplantes. Para compreender melhor as alterações microcirculatórias nos pulmões de doadores com MC, o presente estudo investigou a microcirculação pulmonar em um modelo de roedor com MC via microscopia intravital. MÉTODOS: Ratos Wistar machos foram anestesiados e ventilados mecanicamente. Eles foram submetidos a trepanação e a MC induzida por meio do aumento da pressão intracraniana. Os ratos do grupo Sham (SH), utilizado como controle, foram submetidos apenas à trepanação. Em ambos os grupos, foram monitorados o O2 expiratório e o CO2, e, após 3 horas, foi realizada a toracotomia e criada uma janela para observar a superfície pulmonar usando o sistema de microscopia intravital. As expressões pulmonares das moléculas de adesão intercelular (ICAM)-1 e da óxido nítrico-sintase endotelial (eNOS) foram avaliadas por imuno-histoquímica, e as citocinas foram medidas em amostras pulmonares. RESULTADOS: Três horas após os procedimentos cirúrgicos, a perfusão pulmonar foi de 73% no grupo SH. Por outro lado, os animais com MC apresentaram uma importante diminuição na perfusão do órgão para 28% (p = 0,036). O comprometimento microcirculatório pulmonar após a indução de MC foi associado a um aumento do número de leucócitos recrutados para o tecido pulmonar, além de uma redução na expressão de eNOS e um aumento na expressão de ICAM-1 nas células endoteliais do pulmão. Os ratos com MC apresentaram valores mais elevados de O2 expiratório e valores mais baixos de CO2 em comparação com os animais SH após 3 horas de monitorização. CONCLUSÕES: Os dados apresentados demonstraram que a MC desencadeia uma importante hipoperfusão e inflamação nos pulmões, comprometendo a microcirculação pulmonar do doador.
Assuntos
Morte Encefálica/fisiopatologia , Células Endoteliais , Pulmão/irrigação sanguínea , Microcirculação/fisiologia , Doadores de Tecidos , Animais , Masculino , Microvasos , Modelos Teóricos , Ratos , Ratos WistarRESUMO
OBJECTIVE: The study objective was to evaluate the effect of bilateral sympathectomy on ventricular remodeling and function in a rat model of dilated cardiomyopathy induced by doxorubicin. METHODS: Dilated cardiomyopathy was induced in male Wistar rats by weekly intraperitoneal injection of doxorubicin (2 mg/kg) for 9 weeks. Animals were divided into 4 groups: dilated cardiomyopathy; bilateral sympathectomy, submitted on day 15 of the protocol to bilateral sympathectomy; angiotensin-converting enzyme inhibitor, treated with enalapril through day 15 until the end of the experimental protocol; and sham, nonsubmitted through doxorubicin protocol, with weekly intraperitoneal injections of saline solution (0.9%). The left ventricular function was assessed, and the heart was collected for posterior analyses. RESULTS: The dilated cardiomyopathy group presented a significant decrease in the myocardial efficiency when compared with the sham group (33.4% vs 71.2%). Only the bilateral sympathectomy group was able to preserve it (57.5%; P = .0001). A significant dilatation in the left ventricular chamber was observed in the dilated cardiomyopathy group (15.9 µm2) compared with the sham group (10.2 µm2; P = .0053). Sympathectomy and enalapril prevented ventricular remodeling (9.5 and 9.6 µm2, respectively; P = .0034). There was a significant increase in interstitial myocardial fibrosis in the dilated cardiomyopathy group (14.8%) when compared with the sham group (2.4%; P = .0001). This process was significantly reduced with sympathectomy and enalapril (8.7 and 3.9%, respectively; P = .0001). CONCLUSIONS: Bilateral sympathectomy was effective in preventing remodeling and left ventricular dysfunction in a rat model of dilated cardiomyopathy induced by doxorubicin.
Assuntos
Cardiomiopatia Dilatada/induzido quimicamente , Cardiomiopatia Dilatada/cirurgia , Doxorrubicina/toxicidade , Simpatectomia , Inibidores da Enzima Conversora de Angiotensina/farmacologia , Animais , Modelos Animais de Doenças , Enalapril/administração & dosagem , Humanos , Masculino , Ratos , Ratos Wistar , Disfunção Ventricular Esquerda/prevenção & controle , Remodelação VentricularRESUMO
OBJECTIVES: To evaluate the influence of bilateral or left sympathectomy on left ventricular remodeling and function after myocardial infarction in rats. METHODS: Myocardial infarction was induced in rats by ligation of the left anterior descending coronary. Seven days later, rats were divided into 4 groups: the myocardial infarction, myocardial infarction with left sympathectomy, myocardial infarction with bilateral sympathectomy, and sham groups. After 8 weeks, left ventricular function was evaluated with the use of a pressure-volume conductance catheter under steady-state conditions and pharmacological stress. Infarct size and extracellular matrix fibrosis were evaluated, and cardiac matrix metalloproteinases and myocardial inflammatory markers were analyzed. RESULTS: The myocardial infarction and left sympathectomy group had an increased end diastolic volume, whereas the bilateral sympathectomy group had a mean end-diastolic volume similar to that of the sham group (P < .002). Significant reduction in ejection fraction was observed in the myocardial infarction and left sympathectomy group, whereas it was preserved after bilateral sympathectomy (P < .001). In response to dobutamine, left ventricular contractility increased in sham rats, rising stroke work, cardiac output, systolic volume, end-diastolic volume, ejection fraction, and dP/dt max. Only bilateral sympathectomy rats had significant increases in ejection fraction (P < .001) with dobutamine. Fibrotic tissue and matrix metalloproteinase expression decreased in the bilateral sympathectomy group compared to that in the myocardial infarction group (P < .001) and was associated with left ventricular wall thickness maintenance and better apoptotic markers in noninfarcted myocardium. CONCLUSIONS: Bilateral sympathectomy effectively attenuated left ventricular remodeling and preserved systolic function after myocardial infarction induction in rats.
Assuntos
Ventrículos do Coração/inervação , Infarto do Miocárdio/cirurgia , Simpatectomia/métodos , Função Ventricular Esquerda , Remodelação Ventricular , Animais , Apoptose , Catecolaminas/metabolismo , Citocinas/metabolismo , Modelos Animais de Doenças , Fibrose , Ventrículos do Coração/metabolismo , Ventrículos do Coração/patologia , Ventrículos do Coração/fisiopatologia , Masculino , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Contração Miocárdica , Infarto do Miocárdio/metabolismo , Infarto do Miocárdio/patologia , Infarto do Miocárdio/fisiopatologia , Peptídeo Natriurético Encefálico/metabolismo , Ratos Wistar , Recuperação de Função Fisiológica , Volume Sistólico , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
Surgical neuromodulation therapies are still considered a last resort when standard therapies have failed for patients with progressive heart failure (HF). Although a number of experimental studies have provided robust evidence of its effectiveness, the lack of strong clinical evidence discourages practitioners. Thoracic unilateral sympathectomy has been extensively studied and has failed to show significant clinical improvement in HF patients. Most recently, bilateral sympathectomy effect was associated with a high degree of success in HF models, opening the perspective to be investigated in randomized controlled clinical trials. In addition, a series of clinical trials showed that bilateral sympathectomy was associated with a decreased risk of sudden death, which is an important outcome in patients with HF. These aspects indicates that bilateral sympathectomy could be an important alternative in the treatment of HF wherein pharmacological treatment barely reaches the target dose.
Assuntos
Humanos , Procedimentos Cirúrgicos Torácicos , Insuficiência Cardíaca/cirurgia , Hiperidrose/cirurgia , Simpatectomia , Resultado do TratamentoRESUMO
OBJECTIVE: Experimental findings support clinical evidence that brain death impairs the viability of organs for transplantation, triggering hemodynamic, hormonal, and inflammatory responses. However, several of these events could be consequences of brain death-associated trauma. This study investigated microcirculatory alterations and systemic inflammatory markers in brain-dead rats and the influence of the associated trauma. METHOD: Brain death was induced using intracranial balloon inflation; sham-operated rats were trepanned only. After 30 or 180 min, the mesenteric microcirculation was observed using intravital microscopy. The expression of Pselectin and ICAM-1 on the endothelium was evaluated using immunohistochemistry. The serum cytokine, chemokine, and corticosterone levels were quantified using enzyme-linked immunosorbent assays. White blood cell counts were also determined. RESULTS: Brain death resulted in a decrease in the mesenteric perfusion to 30%, a 2.6-fold increase in the expression of ICAM-1 and leukocyte migration at the mesentery, a 70% reduction in the serum corticosterone level and pronounced leukopenia. Similar increases in the cytokine and chemokine levels were seen in the both the experimental and control animals. CONCLUSION: The data presented in this study suggest that brain death itself induces hypoperfusion in the mesenteric microcirculation that is associated with a pronounced reduction in the endogenous corticosterone level, thereby leading to increased local inflammation and organ dysfunction. These events are paradoxically associated with induced leukopenia after brain damage.
Assuntos
Morte Encefálica/fisiopatologia , Corticosterona/sangue , Hemodinâmica/fisiologia , Mediadores da Inflamação/sangue , Circulação Esplâncnica/fisiologia , Animais , Modelos Animais de Doenças , Molécula 1 de Adesão Intercelular/fisiologia , Leucopenia/sangue , Leucopenia/etiologia , Masculino , Microcirculação/fisiologia , Microscopia de Fluorescência , Selectina-P/fisiologia , Distribuição Aleatória , Ratos , Ratos WistarRESUMO
PURPOSE: Bacterial translocation has been shown to occur in critically ill patients after extensive trauma, shock, sepsis, or thermal injury. The present study investigates mesenteric microcirculatory dysfunctions, the bacterial translocation phenomenon, and hemodynamic/metabolic disturbances in a rat model of intestinal obstruction and ischemia. METHODS: Anesthetized (pentobarbital 50 mg/kg, i.p.) male Wistar rats (250-350 g) were submitted to intestinal obstruction or laparotomy without intestinal obstruction (Sham) and were evaluated 24 hours later. Bacterial translocation was assessed by bacterial culture of the mesenteric lymph nodes (MLN), liver, spleen, and blood. Leukocyte-endothelial interactions in the mesenteric microcirculation were assessed by intravital microscopy, and P-selectin and intercellular adhesion molecule (ICAM)-1 expressions were quantified by immunohistochemistry. Hematocrit, blood gases, lactate, glucose, white blood cells, serum urea, creatinine, bilirubin, and hepatic enzymes were measured. RESULTS: About 86% of intestinal obstruction rats presented positive cultures for E. coli in samples of the mesenteric lymph nodes, liver, and spleen, and 57% had positive hemocultures. In comparison to the Sham rats, intestinal obstruction induced neutrophilia and increased the number of rolling (approximately 2-fold), adherent (approximately 5-fold), and migrated leukocytes (approximately 11-fold); this increase was accompanied by an increased expression of P-selectin (approximately 2-fold) and intercellular adhesion molecule-1 (approximately 2-fold) in the mesenteric microcirculation. Intestinal obstruction rats exhibited decreased PaCO2, alkalosis, hyperlactatemia, and hyperglycemia, and increased blood potassium, hepatic enzyme activity, serum urea, creatinine, and bilirubin. A high mortality rate was observed after intestinal obstruction (83% at 72 h vs. 0% in Sham rats). CONCLUSION: Intestinal obstruction and ischemia in rats is a relevant model for the in vivo study of mesenteric microcirculatory dysfunction and the occurrence of bacterial translocation. This model parallels the events implicated in multiple organ dysfunction (MOD) and death.
Assuntos
Translocação Bacteriana/fisiologia , Escherichia coli/fisiologia , Obstrução Intestinal/fisiopatologia , Intestino Delgado/irrigação sanguínea , Isquemia/fisiopatologia , Microcirculação/fisiologia , Animais , Biomarcadores/sangue , Modelos Animais de Doenças , Imuno-Histoquímica , Obstrução Intestinal/sangue , Obstrução Intestinal/microbiologia , Intestino Delgado/microbiologia , Intestino Delgado/fisiopatologia , Masculino , Insuficiência de Múltiplos Órgãos/fisiopatologia , Ratos , Ratos WistarRESUMO
OBJECTIVE: Experimental findings support clinical evidence that brain death impairs the viability of organs for transplantation, triggering hemodynamic, hormonal, and inflammatory responses. However, several of these events could be consequences of brain death-associated trauma. This study investigated microcirculatory alterations and systemic inflammatory markers in brain-dead rats and the influence of the associated trauma. METHOD: Brain death was induced using intracranial balloon inflation; sham-operated rats were trepanned only. After 30 or 180 min, the mesenteric microcirculation was observed using intravital microscopy. The expression of Pselectin and ICAM-1 on the endothelium was evaluated using immunohistochemistry. The serum cytokine, chemokine, and corticosterone levels were quantified using enzyme-linked immunosorbent assays. White blood cell counts were also determined. RESULTS: Brain death resulted in a decrease in the mesenteric perfusion to 30 percent, a 2.6-fold increase in the expression of ICAM-1 and leukocyte migration at the mesentery, a 70 percent reduction in the serum corticosterone level and pronounced leukopenia. Similar increases in the cytokine and chemokine levels were seen in the both the experimental and control animals. CONCLUSION: The data presented in this study suggest that brain death itself induces hypoperfusion in the mesenteric microcirculation that is associated with a pronounced reduction in the endogenous corticosterone level, thereby leading to increased local inflammation and organ dysfunction. These events are paradoxically associated with induced leukopenia after brain damage.
Assuntos
Animais , Masculino , Ratos , Morte Encefálica/fisiopatologia , Corticosterona/sangue , Hemodinâmica/fisiologia , Mediadores da Inflamação/sangue , Circulação Esplâncnica/fisiologia , Modelos Animais de Doenças , Molécula 1 de Adesão Intercelular/fisiologia , Leucopenia/sangue , Leucopenia/etiologia , Microscopia de Fluorescência , Microcirculação/fisiologia , Selectina-P/fisiologia , Distribuição Aleatória , Ratos WistarRESUMO
PRUPOSE: Bacterial translocation has been shown to occur in critically ill patients after extensive trauma, shock, sepsis, or thermal injury. The present study investigates mesenteric microcirculatory dysfunctions, the bacterial translocation phenomenon, and hemodynamic/metabolic disturbances in a rat model of intestinal obstruction and ischemia. METHODS: Anesthetized (pentobarbital 50 mg/kg, i.p.) male Wistar rats (250-350 g) were submitted to intestinal obstruction or laparotomy without intestinal obstruction (Sham) and were evaluated 24 hours later. Bacterial translocation was assessed by bacterial culture of the mesenteric lymph nodes (MLN), liver, spleen, and blood. Leukocyte-endothelial interactions in the mesenteric microcirculation were assessed by intravital microscopy, and P-selectin and intercellular adhesion molecule (ICAM)-1 expressions were quantified by immunohistochemistry. Hematocrit, blood gases, lactate, glucose, white blood cells, serum urea, creatinine, bilirubin, and hepatic enzymes were measured. RESULTS: About 86 percent of intestinal obstruction rats presented positive cultures for E. coli in samples of the mesenteric lymph nodes, liver, and spleen, and 57 percent had positive hemocultures. In comparison to the Sham rats, intestinal obstruction induced neutrophilia and increased the number of rolling (~2-fold), adherent (~5-fold), and migrated leukocytes (~11-fold); this increase was accompanied by an increased expression of P-selectin (~2-fold) and intercellular adhesion molecule-1 (~2-fold) in the mesenteric microcirculation. Intestinal obstruction rats exhibited decreased PaCO2, alkalosis, hyperlactatemia, and hyperglycemia, and increased blood potassium, hepatic enzyme activity, serum urea, creatinine, and bilirubin. A high mortality rate was observed after intestinal obstruction (83 percent at 72 h vs. 0 percent in Sham rats). CONCLUSION: Intestinal obstruction and ischemia in rats is a relevant model for ...
Assuntos
Animais , Masculino , Ratos , Translocação Bacteriana/fisiologia , Escherichia coli/fisiologia , Obstrução Intestinal/fisiopatologia , Intestino Delgado/irrigação sanguínea , Isquemia/fisiopatologia , Microcirculação/fisiologia , Biomarcadores/sangue , Modelos Animais de Doenças , Imuno-Histoquímica , Obstrução Intestinal/sangue , Obstrução Intestinal/microbiologia , Intestino Delgado/microbiologia , Intestino Delgado/fisiopatologia , Insuficiência de Múltiplos Órgãos/fisiopatologia , Ratos WistarRESUMO
INTRODUÇÃO: Estudos demonstram que a solução salina hipertônica melhora a hemodinâmica, a microcirculação e modula o sistema imune, atenuando a resposta inflamatória associada ao choque e trauma. Este estudo tem por objetivo avaliar e comparar os efeitos da solução salina hipertônica (NaCl, 7,5%) e do Ringer lactato seguido da ressecção de segmento do intestino necrosado no tratamento da obstrução intestinal e isquemia, através da análise da translocação bacteriana, da disfunção microcirculatória mesentérica, dos distúrbios hemodinâmicos e metabólicos e da disfunção orgânica. MÉTODOS: Ratos Wistar machos (250 300 g) anestesiados (pentobarbital sódico, 50 mg/kg, i.p.) foram submetidos à obstrução intestinal e isquemia (OI, ligadura ao nível do íleo terminal seguida de ligadura de ramos da artéria mesentérica correspondentes à irrigação de 7 10 cm do íleo). Duas horas após os animais foram randomizados em: OI sem tratamento (OI); OI tratado com Ringer lactato (RL, 4 mL/kg, i.v.); e OI tratado com solução salina hipertônica (SH 7,5%, 4 mL/kg, i.v.). Vinte e quatro horas após os procedimentos cirúrgicos iniciais, os ratos obstruídos (OI, RL e SH) foram submetidos à enterectomia. Ratos controles falso-operados (FO) foram submetidos à lapatotomia. Os seguintes parâmetros foram analisados: 1) cultura bacteriana (E. coli) em amostras de linfonodos mesentéricos, fígado, baço e sangue; 2) análise das interações leucócito-endotélio na microcirculação mesentérica por técnica de microscopia intravital; 3) expressão de moléculas de adesão endoteliais (P-selectina e ICAM-1) por imunohistoquímica; 4) quantificação das citocinas e quimiocinas CINC-1 e CINC-2 no soro por enzimaimunoensaio; 5) histologia intestinal; 6) bioquímica sérica; 7) gasometria, hematócrito, lactato, glicose e leucograma; 8) insulina e corticosterona e; 9) sobrevida. RESULTADOS: O tratamento com SH reduziu a bacteremia, a incidência de animais com amostras positivas para E. coli (57%) e a quantidade de...
BACKGROUND: It has been shown that hypertonic saline solution improve hemodynamics, the microcirculation, and modulate the immune system, attenuating the inflammatory response associated with shock and trauma. The present study aims to investigate and compare the effects of hypertonic saline solution (NaCl, 7.5%) and lactated Ringer´s solution in the treatment of intestinal obstruction and ischemia, analysing the bacterial translocation phenomenon, mesenteric microcirculatory dysfunctions, hemodynamic/metabolic disturbances, and organ dysfunction in a rat model of intestinal obstruction and ischemia (IO) followed by resection of the necrotic small bowel segment. METHODS: Anesthetized (pentobarbital 50 mg/kg, i.p.) male Wistar rats (250-300 g) were submitted to IO (ligature at the level of the terminal ileum, followed by ligation of mesenteric vessels that supply 7 10 cm of the ileal loop). Two hours thereafter animals were randomized into: IO without treatment; IO treated with lactated Ringer´s (LR, 4 ml/kg, i.v.) solution; IO treated with hypertonic saline (HS, 7.5%, 4 mL/kg, i.v.) solution. Twenty-four hours thereafter, IO rats (IO, LR, HS) were submitted to enterectomy. Control Sham-operated rats were submitted to laparotomy only. The following parameters were analysed: 1) bacterial cultures (E. coli) from mesenteric lymph nodes, liver, spleen, and blood samples; 2) analyses of leukocyte-endothelial interactions at the mesenteric microcirculation by intravital microscopy; 3) expression of endothelial adhesion molecules (P-selectin, ICAM-1) by immunohistochemistry; 4) quantification of serum cytokines and chemokines (CINC-1, CINC-2) by enzyme-linked immunosorbent assay; 5) intestinal histology; 6) serum biochemistry; 7) blood gases, hematocrit, lactate, glycemia, white blood cell counts; 8) serum insulin and corticosterone; 9) survival rate. RESULTS: Treatment with HS reduced the number of animals with positive samples for the presence of E. coli (57%)...