Pentoxifylline attenuates leukocyte-endothelial interactions in a two-hit model of shock and sepsis.

BACKGROUND: This study investigated the effects of pentoxifylline (PTX) combined with resuscitation fluids on microcirculatory dysfunctions in a two-hit model of shock and sepsis. MATERIALS AND METHODS: Male Wistar rats (250 g) were submitted to hemorrhagic shock and reperfusion followed by sepsis induced by cecal ligation and puncture. For the initial treatment of shock, rats were randomly divided into: sham, no injury, no treatment; hypertonic saline solution (HS) (7.5%, 4 mL/kg); lactated Ringer's solution (LR, 3 × shed blood volume); HS + PTX (4 mL/Kg + 25 mg/kg PTX); and LR + PTX (3 × shed blood volume + 25 mg/kg PTX). After 48 h of being exposed to the double injury, leukocyte-endothelial interactions were assessed by intravital microscopy of the mesentery. Endothelial expression of P-selectin and intercellular adhesion molecule-1 (ICAM-1) was evaluated by immunohistochemistry, as well as lung neutrophil infiltration by histology. RESULTS: Lactated Ringer's solution induced marked increases (P < 0.001) in the number of rolling leukocytes per 10 min (two-fold), adherent leukocytes per 100 µm venule length (six-fold), migrated leukocytes per 5000 µm(2) (eight-fold), P-selectin and ICAM-1 expression (four-fold), and lung neutrophil infiltration (three-fold) compared with sham. In contrast, PTX attenuated leukocyte-endothelial interactions, P-selectin and ICAM-1 expression at the mesentery when associated with either LR (P < 0.001) or HS (P < 0.05). Neutrophil migration into the lungs was similarly reduced by PTX (P < 0.05). CONCLUSIONS: Data presented showed that pentoxifylline attenuates microcirculatory disturbances at the mesenteric bed with significant minimization of lung inflammation after a double-injury model of hemorrhagic shock and reperfusion followed by sepsis.

17ß-estradiol and methylprednisolone association as a therapeutic option to modulate lung inflammation in brain-dead female rats.

Introduction: Brain death (BD) is known to compromise graft quality by causing hemodynamic, metabolic, and hormonal changes. The abrupt reduction of female sex hormones after BD was associated with increased lung inflammation. The use of both corticoids and estradiol independently has presented positive results in modulating BD-induced inflammatory response. However, studies have shown that for females the presence of both estrogen and corticoids is necessary to ensure adequate immune response. In that sense, this study aims to investigate how the association of methylprednisolone (MP) and estradiol (E2) could modulate the lung inflammation triggered by BD in female rats. Methods: Female Wistar rats (8 weeks) were divided into four groups: sham (animals submitted to the surgical process, without induction of BD), BD (animals submitted to BD), MP/E2 (animals submitted to BD that received MP and E2 treatment 3h after BD induction) and MP (animals submitted to BD that received MP treatment 3h after BD induction). Results: Hemodynamics, systemic and local quantification of IL-6, IL-1ß, VEGF, and TNF-α, leukocyte infiltration to the lung parenchyma and airways, and adhesion molecule expression were analyzed. After treatment, MP/E2 association was able to reinstate mean arterial pressure to levels close to Sham animals (p<0.05). BD increased leukocyte infiltration to the airways and MP/E2 was able to reduce the number of cells (p=0.0139). Also, the associated treatment modulated the vasculature by reducing the expression of VEGF (p=0.0616) and maintaining eNOS levels (p=0.004) in lung tissue. Discussion: Data presented in this study show that the association between corticoids and estradiol could represent a better treatment strategy for lung inflammation in the female BD donor by presenting a positive effect in the hemodynamic management of the donor, as well as by reducing infiltrated leukocyte to the airways and release of inflammatory markers in the short and long term.

[Initial evaluation of systemic and regional pCO2 gradients as markers of mesenteric hypoperfusion]. / Avaliação inicial dos gradientes sistêmicos e regionais da pCO2 como marcadores de hipoperfusão mesentérica.

BACKGROUND: Mesenteric ischemia is a life-threatening emergency with a mortality rates still ranging between 60% and 100%. AIM: To evaluate the systemic and regional pCO2 gradients changes induced by mesenteric ischemia-reperfusion injury. In addition, we sought to determine if other systemic marker of splanchnic hypoperfusion could detect the initial changes in intestinal mucosal microcirculation after superior mesenteric artery occlusion. METHODS: Seven pentobarbital anesthetized mongrel dogs (20.6 +/- 1.1 kg) were subjected to superior mesenteric artery occlusion for 45 minutes, and followed for an additional 120 minutes. Systemic hemodynamic was evaluated through a Swan-Ganz and arterial catheters, while gastrointestinal tract perfusion by superior mesenteric vein and jejunal serosal blood flows (ultrasonic flowprobe). Intestinal oxygen delivery, extraction and consumption (DO2intest, ERO2intest and VO2intest, respectively), intramucosal pH (gas tonometry), and mesenteric-arterial and mucosal arterial pCO2 gradients (D(vm-a)pCO2 and D(t-a)pCO2, respectively) were calculated. RESULTS: Superior mesenteric artery occlusion was not associated with significant changes on systemic hemodynamics parameters. A significant increase of D(vm-a)pCO2 (1.7 +/- 0.5 to 5.7 +/- 1.8 mm Hg) and D(t-a)pCO2 (8.2 +/- 4.8 to 48.7 +/- 4.6 mm Hg) were detected. During the reperfusion period a significant decrease on DO2intest (67.7 +/- 9.9 to 38.8 +/- 5.3 mL/min) and a compensatory increase on ERO2intest from 5.0 +/- 1.1% to 12.4 +/- 2.7% was observed. CONCLUSION: We conclude that gas tonometry can detect the mesenteric blood flow disturbances sooner than other analyzed parameters. Additionally, we demonstrated that changes on systemic or regional pCO2 gradients are not able to detect the magnitude of intestinal mucosal blood flow reduction after mesenteric ischemia-reperfusion injury.

Mesenteric hypoperfusion and inflammation induced by brain death are not affected by inhibition of the autonomic storm in rats.

OBJECTIVES: Brain death is typically followed by autonomic changes that lead to hemodynamic instability, which is likely associated with microcirculatory dysfunction and inflammation. We evaluated the role of the microcirculation in the hemodynamic and inflammatory events that occur after brain death and the effects of autonomic storm inhibition via thoracic epidural blockade on mesenteric microcirculatory changes and inflammatory responses. METHODS: Male Wistar rats were anesthetized and mechanically ventilated. Brain death was induced via intracranial balloon inflation. Bupivacaine (brain death-thoracic epidural blockade group) or saline (brain death group) infusion via an epidural catheter was initiated immediately before brain death induction. Sham-operated animals were used as controls (SH group). The mesenteric microcirculation was analyzed via intravital microscopy, and the expression of adhesion molecules was evaluated via immunohistochemistry 180 min after brain death induction. RESULTS: A significant difference in mean arterial pressure behavior was observed between the brain death-thoracic epidural blockade group and the other groups, indicating that the former group experienced autonomic storm inhibition. However, the proportion of perfused small vessels in the brain death-thoracic epidural blockade group was similar to or lower than that in the brain death and SH groups, respectively. The expression of intercellular adhesion molecule 1 was similar between the brain death-thoracic epidural blockade and brain death groups but was significantly lower in the SH group than in the other two groups. The number of migrating leukocytes in the perivascular tissue followed the same trend for all groups. CONCLUSIONS: Although thoracic epidural blockade effectively inhibited the autonomic storm, it did not affect mesenteric hypoperfusion or inflammation induced by brain death.

Mesenteric hypoperfusion and inflammation induced by brain death are not affected by inhibition of the autonomic storm in rats

OBJECTIVES: Brain death is typically followed by autonomic changes that lead to hemodynamic instability, which is likely associated with microcirculatory dysfunction and inflammation. We evaluated the role of the microcirculation in the hemodynamic and inflammatory events that occur after brain death and the effects of autonomic storm inhibition via thoracic epidural blockade on mesenteric microcirculatory changes and inflammatory responses. METHODS: Male Wistar rats were anesthetized and mechanically ventilated. Brain death was induced via intracranial balloon inflation. Bupivacaine (brain death-thoracic epidural blockade group) or saline (brain death group) infusion via an epidural catheter was initiated immediately before brain death induction. Sham-operated animals were used as controls (SH group). The mesenteric microcirculation was analyzed via intravital microscopy, and the expression of adhesion molecules was evaluated via immunohistochemistry 180 min after brain death induction. RESULTS: A significant difference in mean arterial pressure behavior was observed between the brain death-thoracic epidural blockade group and the other groups, indicating that the former group experienced autonomic storm inhibition. However, the proportion of perfused small vessels in the brain death-thoracic epidural blockade group was similar to or lower than that in the brain death and SH groups, respectively. The expression of intercellular adhesion molecule 1 was similar between the brain death-thoracic epidural blockade and brain death groups but was significantly lower in the SH group than in the other two groups. The number of migrating leukocytes in the perivascular tissue followed the same trend for all groups. CONCLUSIONS: Although thoracic epidural blockade effectively inhibited the autonomic storm, it did not affect mesenteric hypoperfusion or inflammation induced by brain death. .

Oxygen consumption, pCO2 gradients and regional blood flow distribution in an alternative model of intestinal autotransplantation.

BACKGROUND: Postoperative complications after intestinal transplantation can be attributed to hypothermic storage and reperfusion injury. Our objective was to evaluate, in an alternative model of intestinal autotransplantation, the initial effects of isolated intestinal hypothermic perfusion (at 4 degrees C, IHP) on mucosal and serosal blood flow distribution and correlate these findings with other systemic and regional markers of mesenteric ischemia. In addition, we sought to obtain evidence that intestinal pCO2 measurement can be a useful method for monitoring graft perfusion and early histological changes after small bowel transplantation. MATERIALS AND METHODS: Eight dogs (23.3+/-1.1 kg) were submitted to a in situ IHP for 30 min, followed by a 180-min reperfusion period. Cardiac output, mesenteric vein, and intestinal serosal blood flows (SMVBF and SBF, ultrasonic flowprobe); intestinal mucosal-arterial pCO2 gradient (Dt-apCO2, tonometry); and O2-derived variables were evaluated. RESULTS: IHP induced a reduction in SMVBF (579+/-53 to 321+/-10 mL/min) and SBF, (44.7+/-3.2 to 29.1+/-5.3 mL/min), and an increase in Dt-apCO2 (2+/-2.8 to 20.5+/-4.5 mm Hg). No alterations on systemic metabolic or O2-derived variables were observed. The increase of the Dt-apCO2 correlated with the grade of mucosal injury. CONCLUSION: IHP induces a proportional reduction on blood flow in all layers of the intestine, and none of the systemic markers of splanchnic ischemia predict the intestinal blood flow disturbances during the early phase of intestinal transplantation. In addition, intestinal pCO2 measurement seems to be a useful way for monitoring graft perfusion and histological changes after hypothermic ischemia and reperfusion.

Avaliação inicial dos gradientes sistêmicos e regionais da pCO2 como marcadores de hipoperfusão mesentérica / Initial evaluation of systemic and regional pCO2 gradients as markers of mesenteric hypoperfusion

RACIONAL: Apesar dos recentes avanços nos métodos de imagem e no cuidado dos doentes críticos, a taxa de mortalidade do abdome agudo vascular nas últimas duas décadas continua praticamente inalterada. OBJETIVOS: Avaliar as alterações imediatas dos gradientes regionais da pCO2 induzidas pela isquemia e reperfusão mesentérica. Determinar se outros marcadores sistêmicos de hipoperfusão esplâncnica são capazes de detectar precocemente as alterações circulatórias ocorridas na mucosa intestinal após oclusão da artéria mesentérica superior. MÉTODOS: Foram utilizados sete cães machos sem raça definida (20,6 ± 1,1 kg), submetidos a oclusão da artéria mesentérica superior por 45 minutos, sendo os animais observados por período adicional de 2 horas após a reperfusão. Variáveis hemodinâmicas sistêmicas foram avaliadas por meio de cateter arterial e Swan-Ganz. A perfusão do sistema digestório foi avaliada pela medida do fluxo sangüíneo da veia mesentérica superior e da serosa jejunal (fluxômetro ultra-sônico). Oferta, taxa de extração e consumo intestinal de oxigênio (DO2intest, TEO2intest e VO2intest, respectivamente), pH intramucoso (tonometria a gás) e os gradientes veia mesentérica-arterial e mucosa-arterial da pCO2 (Dvm-a pCO2 e Dt-a pCO2, respectivamente), foram calculados. RESULTADOS: A oclusão da artéria mesentérica superior não esteve associada a alterações hemodinâmicas sistêmicas, mas pôde-se observar aumento significativo do Dvm-a pCO2 (1,7 ± 0,5 para 5,7 ± 1,8 mm Hg) e do Dt-a pCO2 (8,2 ± 4,8 para 48,7 ± 4,6 mm Hg). Na fase de reperfusão observou-se redução da DO2intest (67,7 ± 9,9 para 38,8 ± 5,3 mL/min) e conseqüente aumento da TEO2intest de 5,0 ± 1,1 por cento para 12,4 ± 2,7 por cento. Não houve correlação entre os gradientes da pCO2 analisados. CONCLUSAO: A tonometria permite detectar de maneira precoce a redução de fluxo intestinal. Além disso, pudemos demonstrar que as variações dos gradientes regionais e/ou sistêmicos da pCO2 não são capazes de avaliar a magnitude da redução de fluxo da mucosa intestinal durante o fenômeno de isquemia e reperfusão mesentérica.