Mechanism of the salutary effects of estrogen on kupffer cell phagocytic capacity following trauma-hemorrhage: pivotal role of Akt activation.

Kupffer cells are macrophages in the liver whose major role is to clear circulating pathogens. Decreased phagocytic capacity of Kupffer cells may result in severe systemic infection. We tested the hypothesis that the depressed Kupffer cell phagocytic capacity following trauma-hemorrhage is enhanced by estrogen administration and this occurs due to maintenance of Fc receptor expression and cellular ATP content via the activation of Akt. Male C3H/HeN mice were subjected to sham operation or trauma-hemorrhage and sacrificed 2 h thereafter. Estrogen, with or without an estrogen receptor antagonist (ICI 182,780), a PI3K inhibitor (Wortmannin), or vehicle, was injected during resuscitation. Kupffer cell phagocytic capacity was tested in vivo. The expression of Fc receptors, of Akt phosphorylation, of p38 MAPK phosphorylation, of DNA binding activity of NF-kappaB and ATP content of Kupffer cells were also determined. Trauma-hemorrhage suppressed Kupffer cell phagocytosis by decreasing Fc receptor expression and Akt activation; however, it induced p38 MAPK activation and increased NF-kappaB activity. Cellular ATP levels were also decreased following trauma-hemorrhage. Administration of estrogen following trauma-hemorrhage increased phospho-Akt levels and normalized all the parameters described as well as plasma levels of TNF-alpha, IL-6, and IL-10. Coadministration of ICI 182,780 or Wortmannin abolished the beneficial effects of estrogen in improving the phagocytic capacity of Kupffer cells following trauma-hemorrhage. Thus, activation of Akt plays a crucial role in mediating the salutary effect of estrogen in restoring trauma-hemorrhage-induced suppression of Kupffer cell phagocytosis.

[Effect of Soluvit on stress reaction and leucocyte function in serious burn patients in shock stage].

OBJECTIVE: To investigate the effect of Soluvit on stress reaction and leucocyte function in serious burn patients in shock stage. METHODS: Eighty-seven serious burn patients, who did not undergo operation, were divided into Soluvit treatment group and control group randomly. Patients in Soluvit treatment group were treated with two bottles of Soluvit everyday from postburn 1 st to 14 th day. Patients in control group were given 500 ml normal saline infusion instead. Blood samples were collected for determination of cortisol and malondialdehyde every 4 hours on postburn 1 st day. Leucocyte were isolated for testing chemotaxis distance and phagocytic power on postburn 7 th and 14 th day respectively. RESULTS: The serum cortisol contents in serious burn patients were significantly elevated at 2-4 hours after treatment in Soluvit treatment and control groups. Serum cortisol and malondialdehyde levels were higher than normal values in all serious burn patients in shock stage at 10-12 hours after treatment. But the changes of serum cortisol and malondialdehyde in Soluvit treatment group were all lower than those in control group at 10-48 and 14-48 hours after treatment, respectively (all P<0.05). Leucocyte chemotaxis distance in Soluvit treatment group was longer than that in control group on both postburn 7 th and 14 th day (P<0.05 and P<0.01). In contrast, there were no significant differences in phagocytic power between two groups (both P>0.05). CONCLUSION: Above results suggest that Soluvit can mitigate stress reaction and lipid peroxidation action, but enhance leucocyte chemotaxis function in severe burn patients.

[Use of acetylcysteine and polyoxydonium for the correction of immune homeostasis and antioxidant system impaired by combined action of 1,2-dichloroethane and heavy mechanical trauma].

The experiments on noninbred rats showed that a combination of heavy mechanical trauma and 1,2-dichloroethane in a single dose of 0.5 LD50 leads to summation of the immunosuppressive and prooxidizing effects of both factors. A combined administration of acetylcysteine and polyoxidonium on the background of the combined action of a heavy mechanical trauma and 1, 2-dichloroethane (1.5 LD50) reduces the suppression of immune responses and the activation of lipid peroxidation caused by these factors.

[Influence on the concentration of plasma endotoxin by inhibition of complement activation in traumatic hemorrhagic shock rats].

OBJECTIVE: To investigate the influence on the concentration of plasma endotoxin by inhibition of complement activation in traumatic hemorrhagic shock rats. METHODS: Eighty male SD rats were randomly divided into two groups: control and cobra venom factor (CVF) treatment groups. The hemorrhagic shock induced by trauma was replicated in both groups. The animals were killed preshock and at 1, 6, and 24 hours postresuscitation. Twenty-four hours before hemorrhage, rats were given a mainline dose of either 50 microg/kg CVF or an equal volume of saline solution. The plasma and serum samples were collected at each time point to determine the concentration of endotoxin, the activity of CH50 and diamine oxidase (DAO), and the level of tumor necrosis factor (TNF-alpha) at various time points in two groups. RESULTS: Compared with preshock in control group, serum CH50 levels were decreased promptly at 1 hour postresuscitation. Markedly elevation of the levels of endotoxin and TNF-alpha in blood were found at early time after resuscitation, and they were come rapidly back to the basic level at 6 and 24 hours phase. The activity of DAO in blood was increased significantly at 1 and 6 hours after resuscitation and declined promptly at 24 hours. Compared with the control group, significantly decline of the levels of endotoxin, TNF-alpha and DAO at the various time points after resuscitation were also found in the CVF group. The levels of CH50 in CVF group were always less than 5% during the experiment. CONCLUSION: In traumatic hemorrhagic shock rats CVF pretreatment could decline plasma endotoxin levels by preventing the injury of intestine and gut barrier function, decrease endotoxin translocation and reduce plasma endotoxin levels.

Modulating the Biologic Activity of Mesenteric Lymph after Traumatic Shock Decreases Systemic Inflammation and End Organ Injury.

INTRODUCTION: Trauma/hemorrhagic shock (T/HS) causes the release of pro-inflammatory mediators into the mesenteric lymph (ML), triggering a systemic inflammatory response and acute lung injury (ALI). Direct and pharmacologic vagal nerve stimulation prevents gut barrier failure and alters the biologic activity of ML after injury. We hypothesize that treatment with a pharmacologic vagal agonist after T/HS would attenuate the biologic activity of ML and prevent ALI. METHODS: ML was collected from male Sprague-Dawley rats after T/HS, trauma-sham shock (T/SS) or T/HS with administration of the pharmacologic vagal agonist CPSI-121. ML samples from each experimental group were injected into naïve mice to assess biologic activity. Blood samples were analyzed for changes in STAT3 phosphorylation (pSTAT3). Lung injury was characterized by histology, permeability and immune cell recruitment. RESULTS: T/HS lymph injected in naïve mice caused a systemic inflammatory response characterized by hypotension and increased circulating monocyte pSTAT3 activity. Injection of T/HS lymph also resulted in ALI, confirmed by histology, lung permeability and increased recruitment of pulmonary macrophages and neutrophils to lung parenchyma. CPSI-121 attenuated T/HS lymph-induced systemic inflammatory response and ALI with stable hemodynamics and similar monocyte pSTAT3 levels, lung histology, lung permeability and lung immune cell recruitment compared to animals injected with lymph from T/SS. CONCLUSION: Treatment with CPSI-121 after T/HS attenuated the biologic activity of the ML and decreased ALI. Given the superior clinical feasibility of utilizing a pharmacologic approach to vagal nerve stimulation, CPSI-121 is a potential treatment strategy to limit end organ dysfunction after injury.

Sirtuin 1 Agonist Minimizes Injury and Improves the Immune Response Following Traumatic Shock.

Survival from traumatic injury requires a coordinated and controlled inflammatory and immune response. Mitochondrial and metabolic responses to stress have been shown to play a role in these inflammatory and immune responses. We hypothesized that increases in mitochondrial biogenesis via a sirtuin 1 agonist would decrease tissue injury and partially ameliorate the immunosuppression seen following trauma. C57Bl/6 mice were subjected to a multiple trauma model. Mice were pretreated with either 100 mg/kg per day of the sirtuin 1 agonist, Srt1720, via oral gavage for 2 days prior to trauma and extended until the day the animals were killed, or they were pretreated with peroxisome proliferator-activated receptor γ coactivator 1α (PGC1α) siRNA via hydrodynamic tail vein injection 48 h prior to trauma. Markers for mitochondrial function and biogenesis were measured in addition to splenocyte proliferative capacity and bacterial clearance. Srt1720 was noted to improve mitochondrial biogenesis, mitochondrial function, and complex IV activity following traumatic injury (P < 0.05), whereas knockdown of PGC1α resulted in exacerbation of mitochondrial dysfunction (P < 0.05). These changes in mitochondrial function were associated with altered severity of hepatic injury with significant reductions in serum alanine aminotransferase levels seen in mice treated with srt1720. Splenocyte proliferative capacity and intraperitoneal bacterial clearance were evaluated as markers for overall immune function following trauma-hemorrhage. Treatment with Srt1720 minimized the trauma-induced decreases in splenocyte proliferation (P < 0.05), whereas treatment with PGC1α siRNA led to diminished bacterial clearance. The PGC1α signaling pathway is an important regulator of mitochondrial function and biogenesis, which can potentially be harnessed to protect against hepatic injury and minimize the immunosuppression that is seen following trauma-hemorrhage.

Effects of trauma on immune cell function: impairment of intracellular calcium signaling.

Immunosuppression following injury influences infectious morbidity and mortality. Impaired T-cell activation conceding to inadequate antigen recognition contributes to this immunosuppression. Successful activation and proliferation of T-cells requires precisely specified levels of intracellular calcium thresholds and peak signals. The purpose of this study was to evaluate intracellular calcium signaling following injury. Hospitalized blunt and penetrating trauma patients in a Level 1 Trauma Center following injury and sepsis were tested for immune cell calcium signaling. Peripheral blood mononuclear cells (PBMC) were isolated and calcium signaling tested with Fura-2 AM. PBMC from trauma patients had significantly depressed values of baseline, peak and sustained levels of intracellular calcium prior to and following phytohemagglutinin stimulation when compared to normal controls. This deficit in intracellular calcium signaling is more severe in septic trauma patients (60% reduction). Suppression of calcium signaling appears to be mediated by at least, in part, circulating serum factors. Prostaglandin E2 seems to have a limited contribution to this effect as it is suppressive only when in direct contact with PBMC. Immune cell activation failure can in part be explained by the inadequacy of calcium signaling; restoration of immunocompetence following trauma will have to be addressed by strategies to restore calcium signaling, a vital step necessary for T-cell proliferation following antigen recognition.

Post-traumatic osteomyelitis: analysis of inflammatory cells recruited into the site of infection.

Device-associated infections after implants or endoprostheses inflict local inflammation and ultimately osteolysis, a clinical entity referred to as posttraumatic osteomyelitis. The underlying molecular mechanisms are not yet known; formation of bacterial biofilms on the implant is presumed, conferring resistance to antibiotics and to host defense mechanisms as well. To gain insight into the pathogenesis of post-traumatic osteomyelitis, the infected site was analyzed for the presence of immunocompetent cells. In 18 patients, the infected site was rinsed intraoperatively. This so-called lavage contained 1-2 x 107 leukocytes, predominantly highly activated polymorphonuclear neutrophils (PMNs), as characterized by low expression of CD62L (selectin), and high expression of the adhesion protein CD18, of the high-affinity immunoglobulin (IgG) receptor CD64, and of the LPS-receptor CD14. CD16, the low-affinity IgG receptor, was affected in some patients only. Because the majority of infections were caused by staphylococci species, the effect of bacteria-derived lipoteichoic acid on PMN of healthy donors was tested in vitro. A similar activation pattern was found: rapid down-regulation of CD62L, a slower loss of CD16, and upregulation of CD18, CD64, and CD14. Lipoteichoic acid signaling required p38 mitogen-activated protein kinase and resulted in induction of CD14-specific mRNA and de novo protein synthesis. We conclude that PMNs infiltrate the infected site, but despite local activation they are unable to clear the bacteria, presumably because of biofilm formation. Our data are consistent with the hypothesis that during the ineffective "frustrated" attempt to phagocytose, PMNs release cytotoxic and proteolytic entities that in turn contribute to the progression of tissue injury and ultimately to osteolysis.

Unexpected action of platelet activating factor antagonism after fluid resuscitation from traumatic shock.

BACKGROUND: We have shown that therapy directed at polymorphonuclear neutrophil (PMN) CD18 receptors attenuates sequelae associated with a post-trauma endotoxin (lipopolysaccharide [LPS]) challenge. Platelet activating factor (PAF) stimulates PMNs by CD18-independent pathways, and WEB-2086, a PAF receptor antagonist, blunts septic symptoms in many experimental models. This study tested the hypothesis that the blockade of non-CD18 dependent PMN adherence attenuates trauma- and LPS-evoked pulmonary dysfunction. METHODS: We performed three experiments. First, anesthetized swine were subjected to hind-limb trauma and 30% hemorrhage. After 1 hour animals were resuscitated with shed blood, lactated Ringer's solution (LRS), and WEB-2086 (10 mg/kg/hr) or vehicle. After a 72-hour recovery period, LPS was administered. LPS was then administered without an earlier episode of traumatic shock to animals treated with WEB-2086 or vehicle. Finally, PAF was infused before and after trauma and a dose response curve was obtained. RESULTS: Surprisingly, PAF blockade increased mortality after trauma (5 of 11 WEB-2086 animals versus 1 of 9 vehicle animals; p = 0.15) and depressed cardiac index and O2 delivery at 72 hours (p < 0.05). After LPS administration WEB-2086 treated pigs were unable to manifest the hyperdynamic circulatory compensation seen in the vehicle pigs. In the absence of traumatic shock, WEB-2086 was associated with reduced mortality (four of five WEB-2086 treated pigs versus two of five vehicle pigs survived 5 hours; p = 0.07) and improved arterial PO2 (p = 0.05) and base excess (p = 0.04) 60 minutes after LPS administration. The dose response curve for PAF infusion on the cardiac index was altered after trauma compared with the nontraumatized state. CONCLUSIONS: Because WEB-2086 had unexpected and fundamentally opposite properties before and after trauma, PAF may have a previously undescribed homeostatic role in the compensatory response to injury. These results also suggest that blockade of endogenous inflammatory mediators can have a positive or negative action, depending on the timing of administration and the preexisting condition.

Dextran 70 administration after trauma-hemorrhagic shock does not impair cellular immune functions.

PURPOSE: Although the effects of the colloid dextran 70 on induction of anaphylactoid reactions or reticuloendothelial phagocytosis have been examined previously, its effects on specific cell-mediated immunity after trauma-hemorrhage shock remain unknown. METHODS: Nonheparinized C3H/HeN mice underwent a laparotomy, were bled, and then maintained at a blood pressure of 35 mm Hg for 60 minutes. Then they were resuscitated with either 4 x the shed blood volume as lactated Ringer's solution (LRS) or 2 x LRS + 1 x dextran 70. Control mice underwent all operative protocols but were neither hemorrhaged, nor resuscitated. At 2 or 24 hours posthemorrhage, serum, splenocytes (SPL), and peritoneal macrophages (pM phi, splenic Mo (sM phi) were obtained. Bioassays were used to determine interleukin-2 (IL-2), IL-3, IL-6, and SPL proliferation. RESULTS: Trauma-hemorrhage markedly depressed lymphokine release, splenocyte proliferation, and IL-6 release at 2 hours after the insult. The combination of LRS + dextran did not restore lymphocyte functions, but also did not further suppress them. The release of IL-6 by pM phi and sM phi at 2 and 24 hours after dextran infusion and serum IL-6 remained at the same level as in LRS-treated animals. CONCLUSIONS: The combination of LRS and colloid dextran 70 does not adversely affect ex vivo cell-mediated immune functions during the first 24 hours after its administration after trauma-hemorrhage. Thus, from the immunological standpoint, dextran is a safe resuscitation adjunct.

The gene expression of cytokines and chemokines induced by tourniquet shock in mice.

Traumatic shock is one of the major fields in forensic pathology, but its mechanism remains elusive from the pathophysiological aspects. Tourniquet shock has been established as one of the animal models of traumatic shock, and we examined the gene expression of cytokines and chemokines in the lung and liver in tourniquet shock using mice. Tourniquet was conducted by the application of elastic bands with five turns at both the thighs as high as possible for 2 h, followed by reperfusion. In this procedure, more than 90% mice died within 48 h after reperfusion. Serum hepatic transaminase and hematocrit values significantly increased at 2 h after reperfusion, and their elevation was still evident after 10 h. Histopathologically, hemorrhages, congestion and leukocyte recruitment were observed in the lung and liver specimens after 6 h of reperfusion. Immunohistochemical analysis with anti-myeloperoxidase antibody demonstrated a massive neutrophil infiltration in the lung and liver at 2 h or more after reperfusion. RT-PCR analyses demonstrated that the gene expression of interleukin-1beta, tumor necrosis factor-alpha, monocytes chemoattractant protein-1, macrophage inflammatory protein (MIP)-1alpha, MIP-2, KC and vascular endothelial adhesion molecule-1 was most enhanced in the lung and liver at 2 h after reperfusion. Thus, the gene expression of cytokines and chemokines is presumed to be closely related with the onset of tourniquet shock. From the forensic aspects, these cytokines and chemokines are considered to be useful markers for the early diagnosis of tourniquet shock.

[Escharectomy during burn shock on Th1/Th2 polarization of helper T lymphocytes in rats after thermal injury].

OBJECTIVE: To determine the type-1/type-2 cytokine response of helper T lymphocytes after thermal injury, and to investigate the effects of escharectomy during burn shock stage on the polarization of Th cells. METHODS: One hundred and sixty male Wistar rats were randomized into four groups. In group A, animals were subjected to a 30 percent full-thickness thermal injury without escharectomy. In groups B, C and D, escharectomy and skin allograft were done at 8, 24, and 96 hours postburn, respectively. At 4, 12, 24, 48, 96, 120 and 168 hours postburn, animals were killed and bloods samples as well as spleens were harvested. Enzyme linked immunoadsorbent assay (ELISA) was applied to determine interferon-gamma(IFN-gamma) and interleukin-4(IL-4) levels in blood and spleen tissues. RESULTS: Levels of IFN-gamma and IL-4 rapidly and significantly were increased after scald injury. IFN-gamma levels began to rise within 4 hours postburn, peaking at 24 hours later. IL-4 showed a persistent elevation up to 168 hours postburn, thereby leading to a dominant tendency of Th2 cytokine response on postburn day 7. In group A, all above parameters revealed most obvious changes compared with controls, then ranked in group D, B and C. CONCLUSION: Escharectomy during burn shock stage is helpful to prevent the shift to Th2 cell response after severe thermal injury.

[Microphagal system in inflammatory and infectious processes]. / Mikrofagal'naia sistema pri vospalitel'no-infektsionnykh protsessakh.

The information accrued by now concerning disorders in polymorphonuclear leukocytes (PMNL) and the system of serum factors determining leukotaxis and PMNL bactericidal properties in inflammatory-infectious processes (IIP) of various origin is presented. Generalization of the available information suggests that IIPs develop not in the highest manifestations of the PMNL system phagocytic properties but not infrequently in the presence of its deficiency. The structurally-functionally undisturbed PMNL system favours good health. Disorders in the PMNL system are the structural-functional equivalent of clinical concepts: "reduced defence capacities of the host", "reduced nonspecific antibacterial resistance", etc. Sepsis is characterized by extreme disorders in the PMNL system.

[Sandoglobulin immunocorrection in trauma]. / Immunokorrektsiia sandoglobulinom pri travme.

Twenty one patients with the long-term compression syndrome (LCS) and 12 patients with burns treated with sandoglobulin in combination with antibacterial therapy were followed up. The control groups included 14 and 18 patients, respectively. All the patients had wound infections. Increased or lowered respiratory burst of peripheral blood neutrophils and lowered contents of active T-lymphocytes were detected in the majority of the patients. The patients had also an increased respiratory burst of tissue homogenate in the primary focus. Sandoglobulin decreased the periods of normalization of the immunological indices, body temperature and leukogram shifts to the right. The most pronounced effect of the drug was recorded before radical operations, i.e. in the presence of acute microbial toxemia or in patients with severe and extended burns. The procedure of immunological monitoring developed by the authors rapidly estimates the indications to the use of sandoglobulin alone or in combination with other immunomodulators.

[Prehospital care in traumatic shock]. / Dogospital'naia pomoshch' pri travmaticheskom shoke.

Clinical investigations performed in 735 patients have shown that in addition to the improvement of organisation of the medical aid and the universally recognized resuscitation measures for trauma shock, the prophylactics and treatment of intoxication, metabolic, immune and other disorders developing in the patients soon after trauma should be started as early as at the prehospital step.

[Use of immunomodulator derinat in the treatment of patients with surgical sepsis in traumatic shock]. / Primenenie derinata v lechenii bol'nykh s khirurgicheskim sepsisom pri travme.

The reparative effects of Derinat were studied in 15 patients with traumatic shock whose state was complicated by the development of sepsis. The immunomodulating effect of the drug lies in increasing the number of lymphocytes, reducing the total number of granulocytes and in elevation of the number of functionally valuable cells in them. Using Derinat was shown to facilitate the restoration of the initially reduced number of blood erythrocytes. The maximum effect of the drug was observed within 3-4 days. Readministrations of Derinat should be made twice a week.

[The treatment of soft-tissue wounds by using sorbent bandages]. / Lechenie ran miagkikh tkanei s ispol'zovaniem sorbiruiushchikh poviazok.

On the basis of experimental and clinical researches the authors has found out that the sorption via wound using activated carbonic fibrous material if started at an early post shock period could be considered as an effective method of treatment of severe mechanic injuries with a vast wound surface. This method contributes to the favourable outcome of local wound healing and diminishes the number of complications mainly of purulent-septic nature. The positive influence of sorbent applied directly on wound after surgical treatment consists in promoting tissue metabolism, and reducing the risk of inflammatory processes. There are straight manifestations of general detoxicating effect: diminishing of catabolism processes, reducing of toxic metabolite concentration, stimulating of immune system, falling the rate of toxemia. As a result the number of complications has diminished 40%, and the number of lethal cases--15%.

[The immunologic aspects of inflammatory complications in combined maxillofacial and craniocerebral trauma]. / Immunologicheskie aspekty vospalitel'nykh oslozhnenii pri sochetannoi cheliustno-litsevoi i cherepno-mozgovoi travme.

In 56 patients with combined maxillofacial and craniocerebral trauma the indices of cellular and humoral immunity were investigated in the course of acute post-trauma period and related to trauma severity. Investigated was also the influence of ACTH and corticosteroid hormones on the mechanisms of secondary transitory immune deficiency and related complications. It was established that in combined craniocerebral and maxillofacial trauma the late complications developed against the background of a considerable catabolic activation (increased levels of ACTH and cortisol), and suppression of cellular immunity. These were indications to goal-directed immune correction as a part in combined therapeutic intervention at the early stages of traumatic disease.

[Immunopathogenesis of severe wounds and traumas: possibilities of immune correction]. / Immunopatogenez tiazhelykh ranenii i travm: vozmozhnosti immunokorrektsii.

The authors describe the present-day views on the nature of immune dysfunctions in severe traumas. Based on personal clinical experiences and literature data the authors discuss the role of immune dysfunctions in pathogenesis of the traumatic disease. Special attention is given to the role of the immune system in the development of the life-threatening condition: polyorganic insufficiency whose formation mainly results from disorganization and functional failure of the system of immune reactivity. Clinical investigations have shown high effectiveness of early administration for severe wounds and traumas of a new means of immunocorrection--yeast recombinant interleukin-2 of man (preparation Roncoleukin). The administration of this immunocorrector in complex schemes of intensive therapy of the victims was shown to prevent the development of severe pyo-septic pathology and perfectly change the course of the traumatic disease.

[Postaggressive metabolism in severe mechanical trauma]. / Postagressionnyi obmen veshchestv pri tiazheloi mekhanicheskoi travme.

Posttraumatic adaptive reactions are divided by the authors into three periods: the period of "urgent" adaptation (1-2 days), intermediate period (3-7 days) and the period of "long-term" adaptation (after 7 days). The period of "urgent" adaptation may be considered to be more favorable for surgical interventions under conditions of adequate anesthesiological maintenance. A critical associated trauma unlike an isolated one was found to cause more pronounced and continuous strain of the endocrine and metabolic reactions related with the "urgent" adaptation which influences the frequency of complications and lethal outcomes.