RESUMO
Objective: The present study aimed to investigate whether recombinant human erythropoietin (rHuEPO) plays an immunomodulatory function by regulating the TLR4/NF-κB signaling pathway.Materials and methods: C57BL/6 mice were intraperitoneally injected with rHuEPO and, half an hour later, with 50% glycerol at the dose of 7.5 ml/kg to induce crush syndrome (CS)-acute kidney injury (AKI). The levels of TNF-α, IL-1ß, IL-6, serum creatinine (Scr), and creatine kinase (CK) were measured. The kidney tissues were analyzed by HE staining, and macrophage infiltration was detected by immunohistochemistry. Double immunofluorescence staining, RT-qPCR, and western blotting were conducted to analyze TLR4/NF-κB p65 expression. Ferrous myoglobin was co-cultured with RAW264.7 cells to mimic crush injury and the production of proinflammatory cytokines. The expression levels of TLR4 and NF-κB p65 were measured.Results: In vivo study results revealed that rHuEPO ameliorated renal function, tissue damage, production of proinflammatory cytokines, and macrophage infiltration in the kidneys. The protein and mRNA expression levels of genes involved in the TLR4/NF-κB signaling pathway in CS-induced AKI mice were upregulated (p < .05). Meanwhile, the expression levels of TLR4, NF-κB p65, and proinflammatory cytokines in RAW264.7 cells were downregulated in CS-AKI mice injected with rHuEPO (p < .05).Conclusions: Our results demonstrated the immunomodulatory capacity of rHuEPO and confirmed that rHuEPO exerts protective effects against CS-induced AKI by regulating the TLR4/NF-κB signaling pathway in macrophages. Therefore, our findings highlight the therapeutic potential of rHuEPO in improving the prognosis of CS-AKI patients.
Assuntos
Injúria Renal Aguda , Síndrome de Esmagamento , Eritropoetina/farmacologia , Fatores Imunológicos/farmacologia , Macrófagos/imunologia , NF-kappa B/imunologia , Transdução de Sinais/efeitos dos fármacos , Receptor 4 Toll-Like/imunologia , Injúria Renal Aguda/tratamento farmacológico , Injúria Renal Aguda/imunologia , Injúria Renal Aguda/patologia , Animais , Síndrome de Esmagamento/tratamento farmacológico , Síndrome de Esmagamento/imunologia , Síndrome de Esmagamento/patologia , Macrófagos/patologia , Camundongos , Células RAW 264.7 , Proteínas Recombinantes/farmacologia , Transdução de Sinais/imunologiaRESUMO
RATIONALE: Autologous peripheral nerve injury caused by crush syndrome due to alcohol intoxication is relatively rare, and to our knowledge, the compression of 3 upper limb nerves at the same time has not been reported previously. If a compressive peripheral nerve injury is not treated in a timely manner, it is difficult to recover neurological function, and the prognosis is poor. PATIENT CONCERNS: Here, we present a case of a 50-year-old man with ipsilateral radial nerve, median nerve, and ulnar nerve injuries caused by autogenous compression after drunkenness. DIAGNOSIS: Electromyography and nerve conduction studies suggested peripheral nerve injury in the left upper limb. The diagnosis was injury to the radial nerve, median nerve, and ulnar nerve in the left upper arm. INTERVENTIONS: Exploratory neurolysis surgery of the radial nerve, median nerve, and ulnar nerve was performed in the left upper arm. Postoperative oral neurotrophic drugs were administered, and functional exercise was performed. OUTCOMES: After timely diagnosis and treatment, the strength of the left upper arm muscle recovered, and the prognosis of neurological function was satisfactory during 3 years of follow-up sessions. LESSONS: In the treatment of such patients, a comprehensive understanding of their medical history and a strict physical examination should be performed. Combined with neuroelectrophysiological and imaging examination, the diagnosis can be confirmed. After timely diagnosis and treatment, the prognosis is mostly excellent.
Assuntos
Intoxicação Alcoólica/complicações , Síndrome de Esmagamento/etiologia , Nervo Mediano/lesões , Nervo Radial/lesões , Nervo Ulnar/lesões , Intoxicação Alcoólica/patologia , Síndrome de Esmagamento/patologia , Síndrome de Esmagamento/terapia , Humanos , Masculino , Nervo Mediano/patologia , Pessoa de Meia-Idade , Nervo Radial/patologia , Nervo Ulnar/patologiaRESUMO
PURPOSE: Crush syndrome (CS), a serious medical condition characterised by damage to the muscle cells due to pressure, is associated with high mortality, even when patients receive fluid therapy during transit to the hospital or admission to the hospital. There is no standard triage approach for earthquake victims with crush injuries due to the scarcity of epidemiologic and quantitative data. We examined whether mortality can be predicted based on the severity of skin damage so that assess the severity and prognosis in crush syndrome by assessment of skin damage in hairless rats because we have previously observed that CS results in oedema and redness of the skin in rats. METHODS: Anaesthetised rats were subjected to bilateral hind limb compression [1 kg (mild) and 2 kg (severe) loads] with a rubber tourniquet for 5 h. The rats were then randomly divided into three groups: sham, mild CS, and severe CS. RESULTS: The mild and severe CS groups had mortality rates of 20 and 90%, respectively. The severe CS group demonstrated higher rates of hyperkalaemia, hypovolemic shock, acidosis, and inflammation. Skin damage was significantly worse in the severe CS group compared to the mild CS group. Skin damage showed good correlation with pathological severity. CONCLUSIONS: Skin damage is a valid measure of transepidermal water loss and severity of CS. We suggest that these models may be useful to professionals who are not experienced in disaster management to identify earthquake victims at high risk of severe CS.
Assuntos
Síndrome de Esmagamento/diagnóstico , Pele/lesões , Animais , Síndrome de Esmagamento/patologia , Modelos Animais de Doenças , Escala de Gravidade do Ferimento , Masculino , Músculo Esquelético/química , Músculo Esquelético/lesões , Músculo Esquelético/metabolismo , Peroxidase/metabolismo , Prognóstico , Ratos Pelados , Espécies Reativas de Oxigênio/metabolismo , Pele/patologiaRESUMO
Rhabdomyolysis is a serious syndrome caused by skeletal muscle injury and the subsequent release of breakdown products from damaged muscle cells into systemic circulation. The muscle damage most often results from strenuous exercise, muscle hypoxia, medications, or drug abuse and can lead to life-threatening complications, such as acute kidney injury (AKI). Rhabdomyolysis and the AKI complication can also occur during crush syndrome, an emergency condition that commonly occurs in victims of natural disasters, such as earthquakes, and man-made disasters, such as wars and terrorism. Myoglobin released from damaged muscle is believed to trigger renal dysfunction in this form of AKI. Recently, macrophages were implicated in the disease pathogenesis of rhabdomyolysis-induced AKI, but the precise molecular mechanism remains unclear. In the present study, we show that macrophages released extracellular traps (ETs) comprising DNA fibers and granule proteins in a mouse model of rhabdomyolysis. Heme-activated platelets released from necrotic muscle cells during rhabdomyolysis enhanced the production of macrophage extracellular traps (METs) through increasing intracellular reactive oxygen species generation and histone citrullination. Here we report, for the first time to our knowledge, this unanticipated role for METs and platelets as a sensor of myoglobin-derived heme in rhabdomyolysis-induced AKI. This previously unknown mechanism might be targeted for treatment of the disease. Finally, we found a new therapeutic tool for prevention of AKI after rhabdomyolysis, which might rescue some sufferers of this pathology.
Assuntos
Injúria Renal Aguda/genética , Síndrome de Esmagamento/genética , Ativação Plaquetária/genética , Rabdomiólise/genética , Injúria Renal Aguda/etiologia , Injúria Renal Aguda/patologia , Animais , Citrulinação/genética , Síndrome de Esmagamento/etiologia , Síndrome de Esmagamento/patologia , DNA/genética , DNA/metabolismo , Modelos Animais de Doenças , Armadilhas Extracelulares/genética , Armadilhas Extracelulares/metabolismo , Heme/metabolismo , Histonas/metabolismo , Humanos , Macrófagos/metabolismo , Macrófagos/patologia , Camundongos , Músculo Esquelético/lesões , Músculo Esquelético/patologia , Mioglobina/genética , Espécies Reativas de Oxigênio/metabolismo , Rabdomiólise/complicações , Rabdomiólise/patologia , Vesículas Secretórias/genéticaRESUMO
BACKGROUND: Inflammation plays a crucial role in kidney damage after crush syndrome (CS). Several researchers report that high mobility group box-1 protein (HMGB1) may be the vital trigger in kidney damage, and tumor necrosis factor-α (TNF-α) and c-Jun N-terminal kinase (JNK) are involve in this pathophysiological process, but their biological roles are unclear. This study aimed to explore the relationship between HMGB1, JNK, and TNF-α in kidney damage. METHODS: The crush injury model was established using weight compression. The reliability of the crush injury model was determined by hematoxylin-eosin (HE) staining. Western blot was used to detect the expression of HMGB1, JNK, and TNF-α, and TUNEL was used to mark apoptotic cells in the renal cortex. RESULTS: The results showed that the highest expression of HMGB1 in muscle was 12 h after CS. JNK and TNF-α increased and peaked at 1 day after CS in kidneys. Western blot analysis revealed that anti-HMGB1 antibody could downregulate the expression of JNK and TNF-α. Anti-TNF-α could downregulate activation of JNK, and SP600125 could downregulate expression of TNF-α in the kidneys. In addition, anti-HMGB1 antibody, anti-TNF-α antibody, and SP600125 could reduce cellular apoptosis in the renal cortex. CONCLUSIONS: It is possible that JNK and TNF-α commonly contribute to kidney damage by assembling a positive feedback cycle after CS, leading to increased apoptosis in the renal cortex. HMGB1 from the muscle may be the trigger.
Assuntos
Síndrome de Esmagamento/metabolismo , Proteína HMGB1/metabolismo , Rim/patologia , Sistema de Sinalização das MAP Quinases , Fator de Necrose Tumoral alfa/metabolismo , Animais , Antracenos , Síndrome de Esmagamento/mortalidade , Síndrome de Esmagamento/patologia , Proteína HMGB1/antagonistas & inibidores , Marcação In Situ das Extremidades Cortadas , Rim/metabolismo , Masculino , Camundongos Endogâmicos C57BL , Músculo Esquelético/metabolismo , Músculo Esquelético/patologiaRESUMO
BACKGROUND: We produced fibroblast growth factor (FGF)-2-containing low-molecular-weight heparin (Fragmin)/protamine nanoparticles (FGF-2 + F/P NPs). The purpose of this study was to evaluate the effectiveness of the local administration of FGF-2 + F/P NPs on repairing crush syndrome (CS)-injured lesions after compression release using a nonlethal and reproducible CS injury rat model. MATERIALS AND METHODS: The hind limbs of the anesthetized rats were compressed for 6 h using 3.6 kg blocks, as previously described. The effects of administering FGF-2 + F/P NPs (group A), F/P NPs alone (group B), FGF-2 alone (group C), and saline (control; group D) were examined. Motor function, surface blood flow in the hind limbs, and the wet/dry weight ratio in the tibialis anterior muscle were examined for 1-28 d after the compression release. Histologic analyses were also performed. RESULTS: At the middle and late stages (3-28 d after the compression release), group A had higher scores in the motor function, improved blood flow, increased number of blood vessels, and faster recovered muscle tissue, compared with the other groups. There was no significant difference in enhanced edema in the tibialis anterior muscle among all groups. CONCLUSIONS: The local administration of FGF-2 + F/P NPs to a CS-injured lesion was effective in repairing damaged muscle tissue after compression release.
Assuntos
Anticoagulantes/administração & dosagem , Síndrome de Esmagamento/tratamento farmacológico , Dalteparina/administração & dosagem , Fator 2 de Crescimento de Fibroblastos/administração & dosagem , Neovascularização Fisiológica/efeitos dos fármacos , Protaminas/administração & dosagem , Animais , Síndrome de Esmagamento/patologia , Modelos Animais de Doenças , Avaliação Pré-Clínica de Medicamentos , Quimioterapia Combinada , Edema/prevenção & controle , Injeções Intralesionais , Locomoção/efeitos dos fármacos , Extremidade Inferior/irrigação sanguínea , Extremidade Inferior/patologia , Masculino , Nanopartículas/administração & dosagem , Ratos Sprague-Dawley , Fluxo Sanguíneo Regional/efeitos dos fármacosRESUMO
BACKGROUND: Forklift-related crush injuries of the foot and ankle are relatively common in cities with shipping and construction industries. There is a paucity of literature on the incidence and sequelae of such injuries. We aimed to describe the incidence, patterns of injuries, sequelae, and morbidity associated with this type of injury. METHODS: A retrospective review of all patients with forklift-related crush injuries of the foot and ankle for 4 years was conducted. Patients' demographics, mechanisms and patterns of injury, fracture type, compartment syndrome, number of reconstructive operations, operative details, length of hospital stay, medical leave, repeat evaluation in emergency room, and complications were recorded and analyzed. RESULTS: There were 113 (2.17%) patients with forklift-related crush injuries out of 5209 patients seen in our institution for injuries of the foot and ankle. Crush injury from the wheels of the forklift truck was the most common mechanism at 71 (62.8%) patients. The forefoot was the most commonly injured region, followed by the midfoot, hindfoot, and ankle, with almost one-third (28.3%) of the patients having multiple injuries to the foot. Nine (8%) had open fractures, while 5 (4.4%) had compartment syndromes. Forty (35.4%) patients required hospitalization, and 35 (87.5%) of those hospitalized required operative intervention. Those who had surgery were more likely to have complications compared with those who did not require operative intervention (16 [45.7%] of 35 patients vs 7 [9%] of 78 patients; P < .05) and more likely to require longer medical leave (mean, 183 vs 30 days, P < .05). CONCLUSION: Forklift-related crush injuries of the foot and ankle are increasingly common in industrialized cities. The forefoot is commonly affected with involvement of multiple regions. Up to one-third of affected patients required hospitalization and multiple operative interventions resulting in loss of productivity, income, and significant morbidity. The possibility of residual disabilities must be clearly defined to the patients and their employers to manage potential workplace limitations and long-term expectations. LEVEL OF EVIDENCE: Level IV retrospective case series.
Assuntos
Acidentes de Trabalho/estatística & dados numéricos , Traumatismos do Tornozelo/epidemiologia , Síndrome de Esmagamento/epidemiologia , Traumatismos do Pé/epidemiologia , Adulto , Idoso , Traumatismos do Tornozelo/patologia , Traumatismos do Tornozelo/cirurgia , Síndrome de Esmagamento/patologia , Síndrome de Esmagamento/cirurgia , Feminino , Traumatismos do Pé/patologia , Traumatismos do Pé/cirurgia , Fraturas Ósseas/epidemiologia , Fraturas Ósseas/patologia , Fraturas Ósseas/cirurgia , Humanos , Incidência , Escala de Gravidade do Ferimento , Tempo de Internação , Masculino , Pessoa de Meia-Idade , Procedimentos Ortopédicos , Estudos Retrospectivos , Adulto JovemRESUMO
OBJECTIVE: To explore the pathogenic mechanism of myoglobin-induced endoplasmic reticulum stress (ERS) and apoptosis in tubular epithelial cells in acute kidney injury (AKI) mouse model of crush syndrome. METHODS: Eighteen C56BL/6 mice were randomly divided into control group, modeling 8 h group and modeling 24 h group. The AKI model of crush syndrome was established by intramuscular injection of 50% glycerol saline solution into thigh (8 microL/g), while equivalent volume of physiological saline was injected in control group. AKI was diagnosed when serum creatinine (sCr) level increased to double value of control group. The mice in the experimental groups were sacrificed at the time points of 8 h and 24 h after injection respectively. Serum Cr was detected and renal tissues was observed under electron microscopy. Apoptosis was detected by TUNEL technique. Marker proteins and mRNA of apoptosis and ERS were detected by immunohistochemistry and real-time PCR. Human kidney proximal tubular cell (HK-2) cells cultured in vitro were randomly divided into control, intervention 6 h and intervention 12 h groups. Control group were incubated in standard cell culture (DMEM/F12) and the two intervention groups were incubated in special DMEM/F12 in which ferrohemoglobin was added. After 6 h and 12 h incubation, the cells were collected and apoptosis was detected by flow cytometry. RESULTS: AKI model of crush syndrome was successfully established, which was proved with sCr doubling at the 8 h after the intramuscular injection of glycerol saline. Swelling of endoplasmic reticulum and mitochondria in proximal tubular epithelial cells was more obvious in the two model groups than that in control group. TUNEL staining showed the percentage of positive cells in AKI groups was higher than that in control group (P<0.05). Immunohistochemistry and real-time PCR showed the expressions of caspase3, caspase12 and CHOP in AKI groups were higher than those in control group (P<0.05). Flow cytometry showed cell apoptosis ratio was higher in model groups than control groiap (P<0.05). CONCLUSION: Myoglobin induced ERS and apoptosis may be involved in the pathogenesis of AKI in crush syndrome.
Assuntos
Injúria Renal Aguda/patologia , Apoptose , Síndrome de Esmagamento/patologia , Estresse do Retículo Endoplasmático , Mioglobina/farmacologia , Animais , Caspase 12/metabolismo , Caspase 3/metabolismo , Linhagem Celular , Creatinina/sangue , Células Epiteliais/patologia , Humanos , Túbulos Renais Proximais/citologia , Camundongos , Camundongos Endogâmicos C57BL , Mitocôndrias/patologia , Fator de Transcrição CHOP/metabolismoRESUMO
Crush syndrome is characterized by prolonged shock resulting from extensive muscle damage and multiple organ failure. However, the pathogenesis of multiple organ failure has not yet been completely elucidated. Therefore, we investigated the molecular biological and histopathological aspects of distant organ injury in crush syndrome by using tourniquet shock model mice. DNA microarray analysis of the soleus muscle showed an increase in the mRNA levels of Cox-2, Hsp70, c-fos, and IL-6, at 3h after ischemia/reperfusion injury at the lower extremity. In vivo staining with hematoxylin and eosin (HE) showed edema and degeneration in the soleus muscle, but no change in the distant organs. Immunohistological staining of the HSP70 protein revealed nuclear translocation in the soleus muscle, kidney, liver, and lung. The c-fos mRNA levels were elevated in the soleus muscle, kidney, and liver, displaying nuclear translocation of c-FOS protein. Terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) analysis suggested the involvement of apoptosis in ischemia/reperfusion injury in the soleus muscle. Apoptotic cells were not found in greater quantities in the kidney. Oxidative stress, as determined using a free radical elective evaluator (d-ROM test), markedly increased after ischemia/reperfusion injury. Therefore, examination of immunohistological changes and determination of oxidative stress are proposed to be useful in evaluating the extent of tourniquet shock, even before changes are observed by HE staining.
Assuntos
Síndrome de Esmagamento/patologia , Insuficiência de Múltiplos Órgãos/patologia , Músculo Esquelético/patologia , Estresse Oxidativo , Traumatismo por Reperfusão/patologia , Animais , Biomarcadores/sangue , Síndrome de Esmagamento/metabolismo , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/metabolismo , Edema/patologia , Proteínas de Choque Térmico HSP70/genética , Proteínas de Choque Térmico HSP70/metabolismo , Imuno-Histoquímica , Marcação In Situ das Extremidades Cortadas , Interleucina-6/genética , Interleucina-6/metabolismo , Rim/metabolismo , Rim/patologia , Fígado/metabolismo , Fígado/patologia , Pulmão/metabolismo , Pulmão/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Modelos Animais , Insuficiência de Múltiplos Órgãos/metabolismo , Músculo Esquelético/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos , Proteínas Proto-Oncogênicas c-fos/genética , Proteínas Proto-Oncogênicas c-fos/metabolismo , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Traumatismo por Reperfusão/metabolismo , Coloração e Rotulagem , Torniquetes , Translocação GenéticaRESUMO
BACKGROUND: The present study aimed to determine whether restraint stress aggravates kidney injury caused by a crush injury through endoplasmic reticulum stress (ERS). METHODS: In this study, Sprague-Dawley rat restraint stress, crush injury, and stressful injury models consisting of restraint stress and crush injury were established. An ERS inhibitor, Salubrinal (Sal), was administered intraperitoneally 30 minutes before induction of daily injury in the stressful injury group. At the end of the experimental procedures, plasma levels of noradrenaline and adrenaline, creatine phosphokinase, creatinine, and blood urea nitrogen were measured. Kidneys were harvested, and paraffin-embedded sections of kidney tissues were processed for hematoxylin-eosin staining and TUNEL assay to verify pathologic changes. Western blot was used to determine the protein levels of glucose-regulated protein 78, CCAAT/enhancer-binding protein-homologous protein, caspase 12, caspase 3, and MCP-1 in kidney specimens. RESULTS: Compared with crush injury, the most significant changes in kidney injury occurred in the stressful injury group, which was inhibited by Sal. The results suggested that restraint stress aggravates kidney injury caused by a crush injury, and the mechanism might involve ERS. Further study showed that double attacks induced a significant increase in the levels of glucose-regulated protein 78, CCAAT/enhancer-binding protein-homologous protein, caspase 12, and caspase 3, which was inhibited by Sal. The same changes were observed using the TUNEL assay. Double attacks also induced an increased expression of the proinflammatory cytokine, MCP-1, which was inhibited by Sal. CONCLUSION: Apoptosis and inflammation induced by ERS are important mechanisms by which restraint stress aggravates kidney injury caused by a crush injury.
Assuntos
Injúria Renal Aguda/metabolismo , Apoptose , Síndrome de Esmagamento/complicações , Citocinas/metabolismo , Estresse do Retículo Endoplasmático/fisiologia , Rim/lesões , Injúria Renal Aguda/etiologia , Injúria Renal Aguda/patologia , Animais , Western Blotting , Creatinina/metabolismo , Síndrome de Esmagamento/metabolismo , Síndrome de Esmagamento/patologia , Modelos Animais de Doenças , Proteínas de Choque Térmico/metabolismo , Marcação In Situ das Extremidades Cortadas , Rim/metabolismo , Rim/patologia , Masculino , Ratos , Ratos Sprague-Dawley , Fator de Transcrição CHOP/metabolismoRESUMO
OBJECTIVE: To establish a repeatable, simple, and effective model of rat crush injury and crush syndrome. METHODS: A total of 42 female Sprague Dawley rats (2-month-old, (CS) so as to lay a foundation for further study on CS. weighing 160-180 g) were divided randomly into the control group (n=6) and experimental group (n=36). The rats of the experimental group were used to establish the crush injury and CS model in both lower limbs by self-made crush injury mould. The survival rate and hematuria rate were observed after decompression. The biochemical indexes of blood were measured at 2, 4, 8, 12, 24, and 48 hours after decompression. The samples of muscle, kidney, and heart were harvested for morphological observation. There was no treatment in the control group, and the same tests were performed. RESULTS: Seven rats died and 15 rats had hematuria during compression in the experimental group. Swelling of the lower limb and muscle tissue was observed in the survival rats after reperfusion. The liver function test results showed that the levels of alanine transaminase and aspartate aminotransferase in the experimental group were significantly higher than those in the control group (P < 0.05). The renal function test results showed that blood urea nitrogen level increased significantly after 2 hours of decompression in the experimental group, showing significant difference when compared with that in the control group at 12, 24, and 48 hours after decompression (P < 0.05); the creatinine level of the experimental group was higher than that of the control group at 4, 8, 12, and 24 hours, showing significant difference at 8, 12, and 24 hours (P < 0.05). The serum K+ concentration of the experimental group was higher than that of the control group at all time, showing significant difference at the other time (P < 0.05) except at 2 hours. The creatine kinase level showed an increasing tendency in the experimental group, showing significant difference when compared with the level of the control group at 4, 8, 12, and 24 hours (P < 0.05). The histological examination of the experimental group showed that obvious edema and necrosis of the muscle were observed at different time points; glomeruli congestion and swelling, renal tubular epithelial cell degeneration, edema, necrosis, and myoglobin tube type were found in the kidneys; and myocardial structure had no obvious changes. CONCLUSION: The method of the crush injury and CS model by self-made crush injury mould is a simple and effective procedure and the experimental result is stable. It is a simple method to establish an effective model of rats crush injury and CS.
Assuntos
Síndrome de Esmagamento/fisiopatologia , Membro Posterior/lesões , Modelos Animais , Alanina Transaminase/sangue , Animais , Aspartato Aminotransferases/sangue , Creatina Quinase/sangue , Síndrome de Esmagamento/sangue , Síndrome de Esmagamento/patologia , Feminino , Rim/patologia , Testes de Função Renal , Miocárdio/patologia , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Reprodutibilidade dos Testes , Taxa de SobrevidaRESUMO
BACKGROUND: Nitrite is an intrinsic signaling molecule with potential therapeutic implications in mammalian ischemia/reperfusion (I/R) injury of the heart, liver, and kidney. Although limb muscle compression and subsequent reperfusion are the causative factors in developing crush syndrome (CS), there has been no report evaluating the therapeutic effects of nitrite on CS. We therefore tested whether nitrite could be a therapeutic agent for the treatment of CS. METHODS: To create a CS model, anesthetized rats were subjected to bilateral hind limb compression with rubber tourniquets for 5 hours, followed by reperfusion for 0 hour to 6 hours while monitoring blood pressure. Saline for the CS group or sodium nitrite (NaNO(2)-100, 200, and 500 µmol/kg) for the nitrite-treated CS groups was intravenously administered immediately before reperfusion. Blood and tissue samples were collected for biochemical analysis. RESULTS: Tissue nitrite levels in injured muscles were significantly reduced in the CS group compared with the sham group during I/R injury. Nitrite administration to CS rats restored nitric oxide bioavailability by enhancing nitrite levels of the muscle, resulting in a reduction of rhabdomyolysis markers such as potassium, lactate dehydrogenase, and creatine phosphokinase. Nitrite treatment also reduced plasma levels of interleukin-6 and myeloperoxidase activities in muscle and lung tissues, finally resulting in a dose-dependent improvement of survival rate from 24% (CS group) to 36% (NaNO(2)-100 group) and 64% (NaNO(2)-200 and 500 groups). CONCLUSION: These results indicate that nitrite reduces I/R-induced muscle damage through its cytoprotective action and contributes to improved survival rate in a rat CS model.
Assuntos
Síndrome de Esmagamento/tratamento farmacológico , Síndrome de Esmagamento/mortalidade , Músculo Esquelético/efeitos dos fármacos , Nitritos/farmacologia , Traumatismo por Reperfusão/tratamento farmacológico , Rabdomiólise/tratamento farmacológico , Análise de Variância , Animais , Determinação da Pressão Arterial , Síndrome de Esmagamento/patologia , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Hemodinâmica/efeitos dos fármacos , Membro Posterior/irrigação sanguínea , Interleucina-6/análise , Interleucina-6/metabolismo , Estimativa de Kaplan-Meier , Masculino , Músculo Esquelético/irrigação sanguínea , Músculo Esquelético/lesões , Peroxidase/sangue , Peroxidase/metabolismo , Distribuição Aleatória , Ratos , Ratos Wistar , Valores de Referência , Traumatismo por Reperfusão/mortalidade , Traumatismo por Reperfusão/fisiopatologia , Rabdomiólise/mortalidade , Rabdomiólise/prevenção & controle , Fatores de Risco , Análise de Sobrevida , Resultado do TratamentoRESUMO
BACKGROUND: Erythropoietin (EPO) is a pleiotropic cytokine with neuroprotective, anti-inflammatory, and muscle regenerative properties. The purpose of our study was to analyze the regenerative capacity of systemically applied EPO in a combined muscle-nerve injury model. METHODS: We performed a crush injury to the left soleus muscle in 84 male Wistar rats. Using an instrumented clamp, the muscle was crushed over its complete length. Simultaneously, the ipsilateral sciatic nerve was sham manipulated or crushed. Upon induction of the trauma, animals received either EPO (E) (single application of 5,000 IU/kg body weight intraperitonial) or vehicle solution (K). After in vivo assessment of mechanical pain according to Frey, thermal hyperalgesia, latency of nerve conduction velocity, and strength of the soleus muscle were analyzed at days 1, 7, and 42 postinjury (n = 7 per group). Cell proliferation and apoptosis were assessed by means of histology and immunohistochemistry. RESULTS: Combined muscle-nerve injury showed a significant loss of muscle strength, which incompletely recovered within 42 days. Rats treated with EPO showed an increased muscle strength after 7 days and 42 days compared with the control group. Pain behavior was highest in the muscle-nerve injured animals at day 7. EPO decreased the pain and increased nerve conduction velocity. Nerve injury diminished proliferation of muscle cells, whereas simultaneous therapy with EPO resulted in a boost of bromdesoxyuridine-positive cells. CONCLUSIONS: EPO promoted muscle restoration and enhanced nerve recovery after combined muscle-nerve injury. Thus, EPO might represent an attractive therapeutic option to optimize the posttraumatic course after injury.
Assuntos
Síndrome de Esmagamento/tratamento farmacológico , Eritropoetina/administração & dosagem , Músculo Esquelético/lesões , Regeneração Nervosa/fisiologia , Traumatismos dos Nervos Periféricos/tratamento farmacológico , Nervo Isquiático/fisiologia , Animais , Síndrome de Esmagamento/patologia , Modelos Animais de Doenças , Membro Posterior/lesões , Imuno-Histoquímica , Injeções Intraperitoneais , Masculino , Força Muscular/efeitos dos fármacos , Força Muscular/fisiologia , Músculo Esquelético/efeitos dos fármacos , Condução Nervosa/efeitos dos fármacos , Traumatismos dos Nervos Periféricos/patologia , Distribuição Aleatória , Ratos , Ratos Wistar , Valores de Referência , Regeneração/efeitos dos fármacos , Regeneração/fisiologia , Nervo Isquiático/efeitos dos fármacos , Nervo Isquiático/patologia , Sensibilidade e EspecificidadeRESUMO
Patients with crush injury often present systemic inflammatory response syndrome and fall into multiple organ failure. The mechanism by which the local tissue damage induces distant organ failure is still unclear. We focused on high-mobility group box 1 protein (HMGB1) as one of the damage-associated molecular pattern molecules that cause systemic inflammation in crush injury. We investigated involvement of HMGB1 and the effects of treatment with anti-HMGB1 antibody in a rat model of crush injury. Both hindlimbs of rats were compressed for 6 h and then released. In the crush injury group, the level of serum HMGB1 peaked at 3 h after releasing compression, followed by the increasing in the serum levels of interleukin 6 and tumor necrosis factor α. Hematoxylin-eosin staining showed substantial damage in the lung 24 h after the crush injury, with upregulation of the expression of receptor for advanced glycation end products, as revealed by immunohistochemical analysis. Intravenous administration of anti-HMGB1 antibody improved survival (n = 20 each group) and significantly suppressed serum levels of HMGB1, interleukin 6, and tumor necrosis factor α compared with the untreated crush injury group (n = 6-9 each group). Histological findings of lung damage were ameliorated, and the expression of receptor for advanced glycation end products was hampered by the treatment. These results indicate that HMGB1 is released in response to damage immediately after crush injury and acts as a proinflammatory mediator. Administration of anti-HMGB1 antibody reduced inflammatory reactions and improved survival by blocking extracellular HMGB1. Thus, HMGB1 appears to be a therapeutic target, and anti-HMGB1 antibody may become a promising novel therapy against crush injury to prevent the progression to multiple organ failure.
Assuntos
Anticorpos Neutralizantes/uso terapêutico , Síndrome de Esmagamento/metabolismo , Proteína HMGB1/metabolismo , Animais , Síndrome de Esmagamento/mortalidade , Síndrome de Esmagamento/patologia , Síndrome de Esmagamento/terapia , Selectina E/sangue , Proteína HMGB1/imunologia , Proteína HMGB1/uso terapêutico , Membro Posterior/lesões , Interleucina-6/sangue , Rim/patologia , Fígado/patologia , Pulmão/patologia , Masculino , Ratos , Ratos Wistar , Receptor para Produtos Finais de Glicação Avançada , Receptores Imunológicos/sangue , Receptor 4 Toll-Like/sangue , Fator de Necrose Tumoral alfa/sangueRESUMO
Crush-syndrom (CS) was characterized by Bywaters E.G.L. in 1941 after London blitz. The soft tissues is followed by acute hemodynamic shock, myoglobinuria, acute renal insufficiency, and lethal endotoxicity. Data of CS pathogenesis study has shown that the largest changes in Crush occur during decompression and are accompanied by acute alteration of brain protein synthesis and strong morphological changes of brain structures. The period of decompression might be characterized by the proteolytic breakdown of the myoglobine and formation of toxic peptides. In our current work we have identified four newly formed peptides in the brain of the animals subjected to the experimental muscle tissue injury. Our investigations related with the CS experimental model have demonstrated that during the 2-hours compression protein synthesis was decreased in cytosol (32,7%) and mitochondria (49%), after 5-h compression there were registered non-significant changes in the level of protein synthesis. Intraperitoneal administration of Proline-rich peptide, ((PRP), 1 mcg/100g weight of rats), originating from proteolysis of C-terminal glycoprotein a neurophysin II along with vasopressin and oxytocin and transferring from the hypothalamus to the neurohypophysis by axonal transport, initiates activation of the protein synthesis in all studied cellular subcomponents of brain cells. The positive effect of the peptide is conditioned, most probably, by activation of the immune system and adaptation mechanisms, including mobilization of endogen-protective mechanisms of the organism.
Assuntos
Síndrome de Esmagamento/metabolismo , Citocinas/farmacologia , Hipotálamo/metabolismo , Peptídeos/farmacologia , Proteínas/metabolismo , Animais , Síndrome de Esmagamento/patologia , Síndrome de Esmagamento/fisiopatologia , Citocinas/química , Citocinas/genética , Humanos , Masculino , Peptídeos/química , Peptídeos/genética , Proteínas/química , Proteínas/genética , Ratos , Ratos WistarRESUMO
BACKGROUND: Post-traumatic inflammatory changes have been identified as major causes of altered organ function and failure. Both hemorrhage and soft tissue damage induce these inflammatory changes. Exposure to heterologous bone in animal models has recently been shown to mimic this inflammatory response in a stable and reproducible fashion. This follow-up study tests the hypothesis that inflammatory responses are comparable between a novel trauma model ("pseudofracture", PFx) and a bilateral femur fracture (BFF) model. MATERIALS AND METHODS: In C57BL/6 mice, markers for remote organ dysfunction and inflammatory responses were compared in four groups (control/sham/BFF/PFx) at the time points 2, 4, and 6 h. RESULTS: Hepatocellular damage in BFF and PFx was highly comparable in extent and evolution, as shown by similar levels of NFkappaB activation and plasma ALT. Pulmonary inflammatory responses were also comparably elevated in both trauma models as early as 2 h after trauma as measured by myeloperoxidase activity (MPO). Muscle damage was provoked in both BFF and PFx mice over the time course, although BFF induced significantly higher AST and CK levels. IL-6 levels were also similar with early and sustained increases over time in both trauma models. CONCLUSIONS: Both BFF and PFx create similar reproducible inflammatory and remote organ responses. PFx will be a useful model to study longer term inflammatory effects that cannot be studied using BFF.
Assuntos
Síndrome de Esmagamento/imunologia , Fraturas do Fêmur/imunologia , Inflamação/imunologia , Traumatismos da Perna/imunologia , Lesões dos Tecidos Moles/imunologia , Lesão Pulmonar Aguda/imunologia , Lesão Pulmonar Aguda/patologia , Animais , Aspartato Aminotransferases/sangue , Creatina Quinase/sangue , Síndrome de Esmagamento/patologia , Modelos Animais de Doenças , Fraturas do Fêmur/patologia , Hemorragia/imunologia , Hemorragia/patologia , Tolerância Imunológica/fisiologia , Inflamação/patologia , Interleucina-6/sangue , Traumatismos da Perna/patologia , Hepatopatias/imunologia , Hepatopatias/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Músculo Esquelético/imunologia , Músculo Esquelético/lesões , Músculo Esquelético/patologia , Lesões dos Tecidos Moles/patologiaRESUMO
To determine differences in clinical outcomes in patients harboring both cubital tunnel syndrome (CuTS) and cervical radiculopathy and the influence of the so-called double crush syndrome. Both procedures were performed in 24 patients, mean age 55 years; first group of 14 patients underwent CuTS surgery as a first procedure. Second group of 10 patients underwent anterior cervical discectomy and fusion (ACDF) then ulnar nerve release (UNR). Two patients underwent bilateral nerve surgery and six multiple cervical discectomies. Surgeries consisted in 26 nerve releases with associated external neurolysis in five, and 34 ACDF procedures, with plating in six. Clinical complaints (mean time 12 months) were sensory in 20 arms, with associated motor weakness and hypothenar atrophy involvement in another six. Electromyography changes were mild (two arms), moderate (16 arms), and severe (eight arms). Mean time of follow-up was 3 years (range 18 months-14 years). Clinical improvement was evidenced in 14 patients. Sensory nerve symptoms improved in 13 limbs in both groups and motor improvement was evident in three patients with UNR as first surgery. A comparative cohort of 20 patients with UNR but without cervical radiculopathy was studied to disclose outcome differences. Of these, 13 patients had clinical improvement. No differences were found among groups. In patients with double crush syndrome, factors that seemed to influence a poor CuTS outcome were evolution of symptoms longer than a year, history of multiple neuropathies or radiculopathies, and ACDF performed before UNR.
Assuntos
Vértebras Cervicais/cirurgia , Síndrome de Esmagamento/cirurgia , Síndrome do Túnel Ulnar/complicações , Síndrome do Túnel Ulnar/cirurgia , Descompressão Cirúrgica , Procedimentos Neurocirúrgicos , Radiculopatia/complicações , Radiculopatia/cirurgia , Adulto , Idoso , Vértebras Cervicais/patologia , Estudos de Coortes , Síndrome de Esmagamento/patologia , Discotomia , Eletrodiagnóstico , Eletromiografia , Feminino , Seguimentos , Humanos , Deslocamento do Disco Intervertebral/cirurgia , Masculino , Pessoa de Meia-Idade , Fusão Vertebral , Resultado do Tratamento , Síndromes de Compressão do Nervo Ulnar/cirurgiaRESUMO
Cells and the extracellular matrix of the loose connective tissue as well as the macrophage system were studied in experimental crush-syndrome in 50 rats using histological, histochemical and electron microscopical methods. The results of the investigation have shown that the components of the loose connective tissue and the macrophage system actively responded to 1-hour-long hindlimb compression. Severe structural and functional injury of the cells and extracellular matrix of the subcutaneous loose connective tissue and of the stroma of different organs (lung, heart, kidney) was detected on dayl, was especially pronounced on day 7 and partly regressed by day 30 after the decompression. These changes included the structural damage of fibroblasts, changes in the number and degranulation of the mast cells, inhibition of RNA synthesis in plasmoblasts and lymphoblasts and the destruction of the collagen, elastic and reticular fibers. The number of macrophages in the liver, lungs, spleen, lymph nodes was reduced on days 1 and 7 after the decompression, in some organs this effect persisted until day 20. The activity of acid phosphatase was reduced in the macrophages in crush-syndrome.
Assuntos
Tecido Conjuntivo/patologia , Síndrome de Esmagamento/patologia , Fibroblastos/patologia , Macrófagos/patologia , Animais , Fígado/patologia , Pulmão/patologia , Linfonodos/patologia , Masculino , Ratos , Baço/patologiaRESUMO
Poor healing of high-energy fractures is often associated with severe muscle damage. This may be partly due to the production, by the injured muscle, of inflammatory cytokines that somehow misdirect bone healing. In order to investigate this question, an animal model was established which embodies a controlled degree of muscle injury with a dose response to the energy absorbed, that can be characterised histologically. Using a custom crush jig, 60 CFLP mice had either 100 or 200 g masses dropped from a fixed height onto the quadriceps muscle, with mechanical measurement of the impact. Energy of impact was reliably and significantly different between the small and large impact conditions, though there was more variability when the large mass was used. Animals were sacrificed at day 2, 4, 8, 16, and 24 post-injury. Muscle histomorphometry at all time points and immunohistochemistry for IL-1beta, IL-6, and TNF-alpha up to day 8 were used as measures of muscle damage, inflammation and repair. Histological sections were analysed into areas of normal muscle fibres, damaged/regenerating muscle fibres and fibrous/inflammatory infiltrate. Early histological response was similar between the two groups; the large crush group displayed significantly greater areas of inflammatory infiltrate and damaged muscle at the later time points after day 8. In the large crush group, IL-1beta and IL-6 expression were significantly higher at day 2 and TNF-alpha was higher at day 8 when compared to the small crush group. The experiment demonstrated that more severe injury to muscle was reliably followed by increased inflammatory cytokine production and a greater degree of inflammation and fibrosis. Increased production of inflammatory cytokines such as TNF-alpha and IL-1beta in the damaged muscles may activate macrophages and recruit fibroblasts, promote scar formation and lead to delayed union or non-union of the adjacent fracture(s).