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INTRODUCTION: To establish an animal model of delayed intravenous resuscitation following seawater immersion after hemorrhagic shock (HS). METHODS: Adult male SD rats were randomly divided into three groups: group NI (HS with no immersion), group SI (HS with skin immersion), and group VI (HS with visceral immersion). Controlled HS in rats was induced by withdrawing 45% of the calculated total blood volume within 30 min. In SI group, immediately after blood loss, 0.5 cm below the xiphoid process was immersed in artificial seawater, at (23 ± 1) °C, for 30 min. In VI group, the rats were performed by laparotomy and the abdominal organs were immersed in (23 ± 1) °C seawater for 30 min. Two hours after seawater immersion, the extractive blood and lactated Ringer's solution were delivered intravenously. The mean arterial pressure (MAP), lactate, and other biological parameters were investigated in different time points. The survival rate of 24 h after HS was recorded. RESULTS: After seawater immersion following HS, MAP and abdominal viscera blood flow decreased significantly, and the plasma levels of lactate and the organ function parameters were increased than the baseline. The above changes in VI group were more serious than those in SI and NI group, especially in myocardial and small intestine damage. The hypothermia, hypercoagulation, and metabolic acidosis were also observed after seawater immersion; the injury was more severely in VI group than that of SI group. However, the plasma levels of sodium, potassium, chlorine, and calcium in VI group were significantly higher than those before injury and in the other two groups. In the VI group, the level of plasma osmolality in instant, 2 h, and 5 h after immersion was 111%, 109%, and 108% of the SI group, respectively, all P < 0.01. The 24-h survival rate of VI group was 25%, which was significantly lower than that of SI group (50%) and NI group (70%), P < 0.05. CONCLUSIONS: The model fully simulated the key damage factors and field treatment conditions, reflected the effects of low temperature and hypertonic damage caused by seawater immersion on the severity and prognosis of naval combat wounds, and provided a practical and reliable animal model for the study of field treatment technology of marine combat shock.
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Choque Hemorrágico , Ratos , Masculino , Animais , Choque Hemorrágico/complicações , Choque Hemorrágico/terapia , Ratos Sprague-Dawley , Modelos Animais de Doenças , Hemorragia , Ressuscitação , Ácido LácticoRESUMO
BACKGROUND: We have recently proved electroacupuncture (EA) ST36 exerted an anti-inflammatory effect in the early phase of intra-abdominal adhesion formation. Evidences indicate that the anti-inflammatory effect of EA ST36 involves a cholinergic anti-inflammatory pathway-dependent mechanism via the vagus nerve. However, the exact effects and accurate vagal modulation of acupuncture in prevention of postoperative intra-abdominal adhesion formation has not been thoroughly evaluated. MATERIALS AND METHODS: Sprague-Dawley rats subjected to abdominal adhesion lesions operation at the cecum and abdominal wall were randomly divided into six groups as follows: (a) EAN: EA non-channel acupoints; (b) EA: EA ST36 after abdominal lesions; (c) VGX/EA: vagotomy (VGX) after abdominal lesions, then EA ST36; (d) VGX/EAN: VGX after abdominal lesions, then EAN; (e) α-BGT/EA: intraperitoneal injection of α-bungarotoxin (α-BGT, an antagonist of α7 subunit of cholinergic nicotinic receptor) before EA ST36, and (f) α-BGT/EAN group: α-BGT injection before EAN. Seven days after abdominal surgical lesions, the levels of tumor necrosis factor-α (TNF-α) and vascular endothelial growth factor (VEGF) in the adhesive tissue were evaluated, macroscopic observation and histopathologic evaluation of adhesion formation and assessment of angiogenesis by immunohistochemical staining of platelet endothelial cell adhesion molecule-1 (CD31) were performed. RESULTS: EA ST36 reduced TNF-α and VEGF levels in adhesive tissue homogenates 7 d after surgery, whereas vagotomy or intraperitoneal injection of α-BGT before EA ST36 reversed its suppressive effects. EA at non-channel acupoints with or without vagotomy or intraperitoneal injection of α-BGT before EA had no suppressive effects on TNF-α and VEGF levels. EA ST36 alleviated the adhesion formation, with both of macroscopic and histopathologic adhesion scores significantly lower than those of the EAN group (1.56 ± 0.29 versus 3.00 ± 0.82, 1.35 ± 0.4 versus 3.91 ± 0.8, respectively, both P < 0.05). Compared with the EAN group, EA ST36 significantly decreased angiogenesis evidenced by reduced CD31 positive microvessel density in adhesive tissue. CONCLUSIONS: EA ST36 might reduce the postoperative local inflammatory response, attenuate the angiogenesis, and alleviate the adhesion formation partly via activating the cholinergic anti-inflammatory mechanism.
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Eletroacupuntura , Aderências Teciduais/prevenção & controle , Técnicas de Fechamento de Ferimentos Abdominais , Animais , Ceco/patologia , Inflamação/metabolismo , Inflamação/prevenção & controle , Masculino , Neovascularização Patológica/metabolismo , Neovascularização Patológica/prevenção & controle , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Distribuição Aleatória , Ratos Sprague-Dawley , Aderências Teciduais/patologia , Fator de Necrose Tumoral alfa/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
The effect on acetabular management in developmental dysplasia of the hip (DDH) patients aged 7 or older with modified low Dega osteotomy procedure was evaluated. Patients between 7 and 14 years old were managed with modified low Dega osteotomy and open reduction and concomitant procedures to evaluate whether low level osteotomy improved the clinical and radiologic outcomes after treatment. Clinical status was assessed using the modified McKay's criteria; radiologic evaluations were assessed for the modified Severin classification, the mean acetabular index (AI), Sharp angle and center-edge (CE) angle. And occurrence of triradiate cartilage injury and complications was recorded. Forty-two DDH patients (57 hips) between 7 and 14 years old were managed with modified low Dega osteotomy. The results demonstrated the latest follow-up 43 hips (75.4%) were rated excellent and 10 hips (17.5%) rated good according to the modified McKay criteria and 41 hips (71.9%) were rated excellent and 11 hips (19.3%) rated good according to Modified Severin classification, respectively. The mean Hip Score improved from 69.53â ±â 7.14 before the operation to 93.17â ±â 8.43 at the final follow-up. The mean AI changed from 31.9° to 20.2°, mean Sharp angle decreased from 59.3° to 38.8° and mean CE angle increased from -10.9° to 35.2°, preoperatively and at latest follow-up, respectively. The modified low Dega osteotomy combined with open reduction and concomitant procedures were found to be adequate in improving instant and sustained clinical and radiographic outcomes for the late detected pediatric walking DDH patients.
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OBJECTIVE: To investigate the effects of PNU282987, a α7 nicotinic acetylcholine receptor agonist (α7nAChR), on organ function and survival rate in dogs with lethal burn shock. METHODS: Twelve adult male Beagle dogs were subjected to 50% total body surface area (TBSA) full-thickness flame injury, and then they were randomly divided into a burn group and a PNU282987 group (PNU group), each n=6. The dogs in PNU group received PNU282987 (0.38 mg/kg, venous pumping) and the dogs in burn group received equal amount of normal saline solution as the control group. The mean arterial pressure (MAP) and the plasma levels of tumor necrosis factor-α (TNF-α), alanine aminotransferase (ALT), MB isoenzyme of creatine kinase (CK-MB), creatinine (Cr), blood urea nitrogen (BUN) were continuously determined before and 0.5, 2, 4, 8, 12, 24 hours after burn. All the above measurements were performed with animals in conscious and cooperative state. At the end of 24-hours-period experiment, the survival rate was recorded. RESULTS: The MAP significantly decreased after burn compared with the baseline data before-injury. The level of MAP in PNU group were significantly higher than those of the burn group from 4 hours after burn, and it returned to 83.6% of baseline level at 24 hours. In contrast, those in the burn group progressively decreased with time till death. The plasma levels of TNF-α in PNU group were significantly lower than those of burn group at each time points post injury. The ALT, Cr, BUN and CK-MB of the burn group increased persistently, while those of the PNU group increased at first and decreased subsequently except for ALT increased persistently, and they were all significantly lower than those of the burn group till to the time point of 12 hours (ALT:51.2±7.0 U/L vs. 104.8±7.4 U/L, Cr:42.7±5.4 µmol/L vs. 88.5±4.8 µmol/L, BUN:4.9±1.2 mmol/L vs. 14.7±1.4 mmol/L, CK-MB:564.0±39.1 U/L vs. 734.0±35.9 U/L, all P<0.05). At the end of 24-hours-period experiment, the survival rate of the PNU group was 50% (3/6) and significantly higher than that of the burn group 0(0/6). CONCLUSIONS: The results indicated that PNU282987 decrease the levels of inflammatory cytokine, improve the organ functions and increase 24-hour survival rate in dogs with lethal burn injury. And PNU282987 may have potential clinical application.
Assuntos
Benzamidas/uso terapêutico , Compostos Bicíclicos com Pontes/uso terapêutico , Queimaduras/tratamento farmacológico , Queimaduras/mortalidade , Choque/tratamento farmacológico , Choque/mortalidade , Animais , Queimaduras/fisiopatologia , Modelos Animais de Doenças , Cães , Masculino , Choque/fisiopatologia , Taxa de SobrevidaRESUMO
BACKGROUND: Surgical resection of the distal stomach impairs gastric emptying. Generally, pylorus and the antrum are removed in the distal gastrectomy, however, the pylorus is removed individually under specific circumstances. We focus on the relation between the pyloric resection and the gastric liquid emptying. AIMS: The present investigation aimed to explore the pylorectomy how to influence gastric liquid emptying in rats. METHODS: Pylorectomy and end-to-end gastroduodenal anastomosis were conducted in rats. Electrodes were implanted in the gastrointestinal serosal surface near the stoma. Total stomach, proximal stomach, distal stomach and duodenal liquid emptying, myoelectricities in the gastrointestinal tract near the stoma, and structures were examined with scintigraphy, electrode recording in vivo, and electron microscopy, respectively. RESULTS: Delayed total stomach and distal stomach emptying were found in pylorectomy rats (p<0.001). However, there was no difference in the proximal stomach and the duodenal liquid emptying compared to the controls (p>0.05). The myoelectricity of 3-5 cpm (cycles/min) in antrum and 10-12 cpm in duodenum were found in the controls and no retrograde or antegrade myoelectricities were recorded in the duodenum and antrum. High-frequency myoelectricities (tachygastria) were recorded in the antrum near the stoma (p<0.01), the retrograde and antegrade myoelectricities propagating through the stoma were recorded, and the regenerated interstitial cells of Cajal were found in stoma under electron microscope observation in pylorectomy rat. CONCLUSIONS: The gastroduodenal incoordination and abnormal myoelectricity related to impaired contraction in the antrum caused the delayed liquid gastric emptying in pylorectomy rats.
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Procedimentos Cirúrgicos do Sistema Digestório/métodos , Esvaziamento Gástrico/fisiologia , Trato Gastrointestinal/fisiopatologia , Piloro/cirurgia , Animais , Eletrodos , Masculino , Modelos Animais , Contração Muscular/fisiologia , Músculo Liso/fisiopatologia , Peristaltismo/fisiologia , Antro Pilórico/fisiopatologia , Ratos , Ratos Sprague-DawleyRESUMO
Gelatinolytic proteinases from common carp dark muscle were purified by 30-60% ammonium sulfate fractionation and a combination of chromatographic steps including ion exchange on DEAE-Sephacel, gel filtration on Sephacryl S-200, ion exchange on High-Q, and affinity on gelatin-Sepharose. The molecular masses of these proteinases as estimated by SDS-PAGE were 75, 67, and 64 kDa under nonreducing conditions. The enzymes revealed high activity at a slightly alkaline pH range, and their activities were investigated using gelatin as substrate. Metalloproteinase inhibitors, EDTA, EGTA, and 1,10-phenanthroline, almost completely suppressed the gelatinolytic activity, whereas other proteinase inhibitors did not show any inhibitory effect. Divalent metal ion Ca (2+) is essential for the gelatinolytic activity. Furthermore, these gelatinolytic proteinases hydrolyze native type I collagen effectively even at 4 degrees C, strongly suggesting their involvement in the texture softening of fish muscle during the post-mortem stage.
Assuntos
Carpas , Gelatinases/isolamento & purificação , Gelatinases/metabolismo , Músculos/enzimologia , Animais , Cromatografia , Colágeno/metabolismo , Inibidores Enzimáticos/farmacologia , Precipitação Fracionada , Gelatina/metabolismo , Gelatinases/química , Concentração de Íons de Hidrogênio , TemperaturaRESUMO
Growth hormone releasing hormone is one of the hormones secreted from the hypothalamus. Because of its potential applications in agriculture and medicine, its short half-life and its expensive chemical synthesis, an analog with high GHRH activity and prolonged half-life was sought after. The fusion partner gene with 127 amino acid residues of the C-terminus from L-asparaginase was recombined with asp-pro-hGHRH(1-44) gene synthesized by PCR method to form one kind of fusion protein with unique acid labile linker Asp-Pro. The Pro-hGHRH(1-44) peptide was purified to homogeneity by means of cell disruption, washing, ethanol precipitation, acid hydrolysis, SP-Sephadex C-25, and Sephadex G-10 column chromatography. The peptide's molecular weight of 5,139 Da as measured by EIS-MS was coincident with the actual values. In the study of the activity, the doses of peptide were 0.1, 1.0, and 10 microg/ml for rat pituitary and 5 microg/ml for human pituitary. The peptide increased GH releases from rat pituitary in a concentration-dependent manner (P<0.05; P<0.01). At 1.0 microg/ml, there was a significant difference between Pro-Pro-hGHRH(1-44)-Gly-Gly-Cys and Pro-hGHRH(1-44) or Pro-Pro-hGHRH(1-44) (P<0.05), whereas the standard hGHRH(1-40) showed no measured rGH release. For human fetal pituitary, the Pro-hGHRH(1-44) peptides showed good GH-releasing activity, but there were no significant differences between them. The structure-activity relationship showed that for both rat and human fetal pituitary, the net GH-releasing activity of the Pro-hGHRH(1-44) analog was more than that of Pro-Pro-hGHRH(1-44). The results of the other hormones from human pituitary showed that the analog had good function-selectivity and species specificity.
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Hormônio Liberador de Hormônio do Crescimento/análogos & derivados , Hormônio Liberador de Hormônio do Crescimento/farmacologia , Animais , Sequência de Bases , Primers do DNA , Humanos , Peptídeos/química , Peptídeos/farmacologia , Hipófise/embriologia , RatosRESUMO
Growth hormone releasing hormone is one of the hormones secreted by the hypothalamus. Because of its potential applications in agriculture and medicine, its short half-life and its expensive chemical synthesis, an analog with high GHRH activity and prolonged half-life has been looked for. The fusion partner gene with 127 amino acid residues of the C-terminus from L-asparaginase was recombined respectively with asp-pro-pro-hGHRH(1-44), asp-pro-hGHRH(1-44) or asp-1pro-GHRH(2-44) genes synthesized by PCR method to form three kinds of fusion proteins with unique acid labile linker Asp-Pro. The Pro-Pro-hGHRH(1-44), Pro-hGHRH(1-44), and 1Pro-GHRH(2-44) peptides were purified to homogeneity by means of cell disruption, washing of inclusion body, ethanol fraction precipitation, acid hydrolysis, SP-Sephadex C-25 and Sephadex G-10 column chromatography. The peptide molecular mass of 5235, 5139 or 4975 Da was determined by ESI mass spectroscopy and purity was determined by SDS-PAGE. In the study of in vitro activity, the antiserum kit against human GH and peptide doses of 0.1, 1.0 and 10 microg/ml were used. These peptides obviously increased GH releases both from human pituitary and from rat pituitary. The activity comparisons showed that there was significant difference between Pro-Pro-hGHRH(1-44)-Gly-Gly-Cys and Pro-Pro-hGHRH(1-44) at 1.0 microg/ml, or between 1Pro-hGHRH(2-44) and Pro-Pro-hGHRH(1-44) or Pro-hGHRH(1-44) at 10 microg/ml. The structure-activity relationships showed that at the original C-terminus, for rat pituitary the activity of the GHRH analog with 1Tyr-->Pro was more than that of Pro-Pro-hGHRH(1-44) or Pro-hGHRH(1-44). The results showed that the analogs had good GH-releasing activity and species specificity.
Assuntos
Hormônio Liberador de Hormônio do Crescimento/análogos & derivados , Hormônio do Crescimento/metabolismo , Hormônio Liberador de Hormônio do Crescimento/genética , Hormônio Liberador de Hormônio do Crescimento/metabolismo , Hormônio Liberador de Hormônio do Crescimento/farmacologia , Hormônios/metabolismo , Humanos , Peso Molecular , Fragmentos de Peptídeos/genética , Fragmentos de Peptídeos/metabolismo , Fragmentos de Peptídeos/farmacologia , Hipófise/metabolismo , Prolina/genética , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/isolamento & purificação , Proteínas Recombinantes de Fusão/metabolismo , Proteínas Recombinantes de Fusão/farmacologia , Relação Estrutura-AtividadeRESUMO
Cartilage lesions are at a high prevalence in dysplastic hips and may relate to arthritic changes and hip joint dysfunction. To date, the effectiveness of repair of articular cartilage defects in the dysplastic hips has not yet been thoroughly evaluated. Here we retrospectively reviewed the effects of acetabuloplasty procedures with/without concomitant autologous tibial periosteal transplantation (ATPT) for articular cartilage defects of the hip in older children with developmental dysplasia of the hip (DDH).Older DDH children with focal cartilage defects of the acetabular or femoral cartilage or both in the hip joint were treated by acetabuloplasty procedures with (Group I) or without (Group II) concomitant ATPT to evaluate the improvements in range of motion (ROM), pain relief of hip, walking tolerability (WL), radiologic evaluations, and outcomes in the long-term follow-up.More satisfactory functional outcome is readily achieved among patients treated with combined acetabuloplasty and ATPT, evidenced by marked pain relief and improved ROM and WL. The latest favorable radiologic evaluation was 70.6% in Group I and 60.0% in Group II, respectively. More hips exhibited congruency between the femoral head and the shell, with less deformity of femoral head and acetabulum or narrowed joint space in Group I. Few major complications were recorded in Group I.Application of periosteal autograft for repair of cartilage defects within the hip joint might be an effective adjunctive treatment for acetabuloplasty in preventing stiffness, reducing pain, and improving ROM and outcomes in hip rehabilitation in the long-term follow-up in older children with DDH.
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Acetábulo/cirurgia , Cartilagem Articular/cirurgia , Cabeça do Fêmur/cirurgia , Luxação Congênita de Quadril/cirurgia , Articulação do Quadril/cirurgia , Periósteo/transplante , Adolescente , Criança , China , Feminino , Seguimentos , Luxação Congênita de Quadril/diagnóstico , Humanos , Masculino , Limitação da Mobilidade , Procedimentos Ortopédicos , Medição da Dor , Complicações Pós-Operatórias/etiologia , Amplitude de Movimento Articular , Estudos RetrospectivosRESUMO
AIM: To investigate whether electroacupuncture ST36 activates enteric glial cells, and alleviates gut inflammation and barrier dysfunction following hemorrhagic shock. METHODS: Sprague-Dawley rats were subjected to approximately 45% total blood loss and randomly divided into seven groups: (1) sham: cannulation, but no hemorrhage; (2) subjected to hemorrhagic shock (HS); (3) electroacupuncture (EA) ST36 after hemorrhage; (4) vagotomy (VGX)/EA: VGX before hemorrhage, then EA ST36; (5) VGX: VGX before hemorrhage; (6) α-bungarotoxin (BGT)/EA: intraperitoneal injection of α-BGT before hemorrhage, then EA ST36; and (7) α-BGT group: α-BGT injection before hemorrhage. Morphological changes in enteric glial cells (EGCs) were observed by immunofluorescence, and glial fibrillary acidic protein (GFAP; a protein marker of enteric glial activation) was evaluated using reverse transcriptase polymerase chain reaction and western blot analysis. Intestinal cytokine levels, gut permeability to 4-kDa fluorescein isothiocyanate (FITC)-dextran, and the expression and distribution of tight junction protein zona occludens (ZO)-1 were also determined. RESULTS: EGCs were distorted following hemorrhage and showed morphological abnormalities. EA ST36 attenuated the morphological changes in EGCs at 6 h, as compared with the VGX, α-BGT and HS groups. EA ST36 increased GFAP expression to a greater degree than in the other groups. EA ST36 decreased intestinal permeability to FITC-dextran (760.5 ± 96.43 ng/mL vs 2466.7 ± 131.60 ng/mL, P < 0.05) and preserved ZO-1 protein expression and localization at 6 h after hemorrhage compared with the HS group. However, abdominal VGX and α-BGT treatment weakened or eliminated the effects of EA ST36. EA ST36 reduced tumor necrosis factor-α levels in intestinal homogenates after blood loss, while vagotomy or intraperitoneal injection of α-BGT before EA ST36 abolished its anti-inflammatory effects. CONCLUSION: EA ST36 attenuates hemorrhage-induced intestinal inflammatory insult, and protects the intestinal barrier integrity, partly via activation of EGCs.
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Eletroacupuntura , Sistema Nervoso Entérico/fisiopatologia , Intestino Delgado/inervação , Neuroglia , Choque Hemorrágico/terapia , Animais , Bungarotoxinas/administração & dosagem , Dextranos/metabolismo , Modelos Animais de Doenças , Sistema Nervoso Entérico/efeitos dos fármacos , Sistema Nervoso Entérico/metabolismo , Fluoresceína-5-Isotiocianato/análogos & derivados , Fluoresceína-5-Isotiocianato/metabolismo , Proteína Glial Fibrilar Ácida/genética , Proteína Glial Fibrilar Ácida/metabolismo , Masculino , Neuroglia/efeitos dos fármacos , Neuroglia/metabolismo , Neuroglia/patologia , Permeabilidade , Ratos Sprague-Dawley , Choque Hemorrágico/genética , Choque Hemorrágico/metabolismo , Choque Hemorrágico/patologia , Choque Hemorrágico/fisiopatologia , Fatores de Tempo , Fator de Necrose Tumoral alfa/metabolismo , Vagotomia , Proteína da Zônula de Oclusão-1/metabolismoRESUMO
Excessive inflammation and high vasopermeability can lead to blood volume loss and tissue edema, which can affect the resuscitation and prognosis for serious burn patients. In this experiment, we investigated the effect of PNU-282987, an α7 nicotine cholinergic receptor agonist on the hemodynamic parameters and survival rate by inhibiting vasopermeability and tissue edema during the fluid resuscitation for lethal burn shock. Forty Beagle dogs with intubation of the carotid artery and jugular vein 24 hours before the injury were subjected to 50% TBSA full-thickness burns, and were randomly divided into following four groups: no resuscitation group (group NR), venous fluid resuscitation group (group R), PNU-282987 treatment group (group P), and fluid resuscitation group plus PNU-282987 group (group RP), with 10 dogs in each group. Hemodynamic variables and biochemical parameters were determined with animals in a conscious and cooperative state. The plasma volume and the vasopermeability were determined by indocyanine green and fluorescein isothiocyanate-dextran, respectively. The level of tumor necrosis factor-α and interleukin-1ß in plasma, and the water content of different organs were also determined. The mean arterial pressure, cardiac output, and plasma volume of all dogs decreased significantly, and the lung extravascular water index and pulmonary vascular permeability index increased remarkably after burn. The hemodynamic parameters deteriorated continually in group N dogs, and then anuria, hyperlactacidemia, and multiple organ dysfunctions developed. The mean arterial pressure and cardiac output of dogs in group R and group RP returned to preinjury levels at 48 hours postburn. The lung extravascular water index and pulmonary vascular permeability in group R were higher than those before preinjury. The dogs in group RP were found to have a significant increase in plasma volume and urine output, and a remarkable decrease in the levels of tumor necrosis factor-α, interleukin-1α, lactic acid, and organ functions compared with those of group R (P <.05). The survival rate of RP group (100%; 10/10) was significantly higher than that of group N (0; 0/10), group P (20%; 2/10), and group R (60%; 6/10). PNU-282987 combined with intravenous fluid resuscitation significantly improved hemodynamics and the survival rate in the early period after this lethal burn shock. The mechanism may be attributable to the lowering of the level of proinflammatory mediators, amelioration of vasopermeability-induced visceral edema, less of blood volume loss, and protection of vital organs through activation of cholinergic anti-inflammatory pathway.
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Benzamidas/farmacologia , Compostos Bicíclicos com Pontes/farmacologia , Queimaduras/complicações , Permeabilidade Capilar/efeitos dos fármacos , Edema/terapia , Agonistas Nicotínicos/farmacologia , Choque/etiologia , Alanina Transaminase/sangue , Animais , Pressão Sanguínea , Água Corporal , Débito Cardíaco , Creatina/sangue , Creatina Quinase Forma MB/sangue , Cães , Edema/etiologia , Interleucina-1beta/sangue , Ácido Láctico/sangue , Pulmão/irrigação sanguínea , Modelos Animais , Volume Plasmático , Distribuição Aleatória , Ressuscitação/métodos , Fator de Necrose Tumoral alfa/sangue , UrinaRESUMO
Burn injury may result in multiple organ dysfunction partially because of apoptotic cell death. The authors have previously shown that valproic acid (VPA) improves survival in a dog burn model. The aim of this study is to examine whether a VPA improves survival in a rodent burn model and whether this was because of inhibition of cell apoptosis. Rats were subjected to third-degree 55% TBSA burns and randomized to treatment with a VPA (300 mg/kg) or normal saline. One group of animals was monitored for 12 hours for survival analysis; another group was killed at 6 hours after injury, and brains, hearts, and blood samples were harvested for examination. Plasma creatine kinase (CK)-MB activities and neuron-specific enolase (NSE) levels were measured to evaluate the cardiac and brain damages. The effects of a VPA on acetylation of histone H3 and caspase-3 activation were also evaluated. Major burn injury resulted in a significant decrease in the acetylation of histone H3, and there was an increase in plasma CK-MB activities, NSE concentrations, and tissue levels of activated caspase-3. A VPA treatment significantly increased the acetylation of histone H3 and survival of the animals after major burn injury. In addition, a VPA treatment significantly attenuated the plasma CK-MB activities, an NSE concentrations, and inhibited caspase-3 activation after major burn injury. These results indicate that a VPA can attenuate cardiac and brain injury, and can improve survival in a rodent model of lethal burn injury. These protective effects may be mediated in part through the inhibition of caspase-3 activation.
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Queimaduras/tratamento farmacológico , Queimaduras/enzimologia , Caspase 3/sangue , Ácido Valproico/farmacologia , Animais , Apoptose , Western Blotting , Creatina Quinase/sangue , Histonas/sangue , Masculino , Fosfopiruvato Hidratase/sangue , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Análise de SobrevidaRESUMO
OBJECTIVE: To explore protective effects of acupuncture at "Zusanli" (ST 36) on cerebral tissue in rats with sepsis. METHODS: Cecal ligation and puncture (CLP) was applied to duplicate the rat model of sepsis. According to random number table, thirty SD rats were divided into a sepsis model group (group A), a sepsis model plus electroacupuncture (EA) group (group B), and a sepsis model plus non-acupoint EA group (group C), ten rats in each one. EA with the same frequency and intensity at "Zusanli" (ST 36) and non-acupoint (0.5 cm laterally to "Zusanli") for 30 min was applied in the group B and group C, respectively. No treatment was given in the goup A. 6 hours after CLP, blood was acquired from abdominal aorta to measure the levels of neuron specific enolase (NSE). Then the rats were sacrificed by abdominal aorta exsanguination to take their cerebral tissue for measuring the levels of tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6). RESULTS: Six hours after CLP, the level of NSE was (3.51 +/- 0.39) ng/mL in group B, which was significantly lower than (7.72 +/- 0.64) ng/mL in group A (P<0.05). The level of NSE was (8.02 +/- 0.72) ng/mL in the group C, which had no statistical significance with group A (P>0.05). The levels of TNF-alpha, IL-6 in cerebral tissue in group B were significantly lower than that of group A and C (all P>0.05). CONCLUSION: EA at "Zusanli" (ST 36) has certain protective effect on septic rat's brain, which has some relationship with decreasing levels of cerebral tissue proinflammatory cytokine and plasma NSE. EA at non-acupoint has no the same action.
Assuntos
Terapia por Acupuntura , Sepse/terapia , Pontos de Acupuntura , Animais , Humanos , Interleucina-6/imunologia , Masculino , Fosfopiruvato Hidratase/imunologia , Ratos , Ratos Sprague-Dawley , Sepse/enzimologia , Sepse/imunologia , Fator de Necrose Tumoral alfa/imunologiaRESUMO
AIM: To investigate whether electroacupuncture (EA) at Zusanli (ST36) prevents intestinal barrier and remote organ dysfunction following prolonged hemorrhagic shock through a vagus anti-inflammatory mechanism. METHODS: Sprague-Dawley rats were subjected to about 45% of total blood volume loss followed by delayed fluid replacement (DFR) with Ringer lactate 3h after hemorrhage. In a first study, rats were randomly divided into six groups: (1) EAN: EA at non-channel acupoints followed by DFR; (2) EA: EA at ST36 after hemorrhage followed by DFR; (3) VGX/EA: vagotomy (VGX) before EA at ST36 and DFR; (4) VGX/EAN: VGX before EAN and DFR; (5) α-bungarotoxin (α-BGT)/EA: intraperitoneal injection of α-BGT before hemorrhage, followed by EA at ST36 and DFR; and (6) α-BGT/EAN group: α-BGT injection before hemorrhage followed by EAN and DFR. Survival and mean arterial pressure (MAP) were monitored over the next 12 h. In a second study, with the same grouping and treatment, cytokine levels in plasma and intestine, organ parameters, gut injury score, gut permeability to 4 kDa FITC-dextran, and expression and distribution of tight junction protein ZO-1 were evaluated. RESULTS: MAP was significantly lowered after blood loss; EA at ST36 improved the blood pressure at corresponding time points 3 and 12 h after hemorrhage. EA at ST36 reduced tumor necrosis factor-α and interleukin (IL)-6 levels in both plasma and intestine homogenates after blood loss and DFR, while vagotomy or intraperitoneal injection of α-BGT before EA at ST36 reversed its anti-inflammatory effects, and EA at ST36 did not influence IL-10 levels in plasma and intestine. EA at ST36 alleviated the injury of intestinal villus, the gut injury score being significantly lower than that of EAN group (1.85 ± 0.33 vs 3.78 ± 0.59, P < 0.05). EA at ST36 decreased intestinal permeability to FITC-dextran compared with EAN group (856.95 ng/mL ± 90.65 ng/mL vs 2305.62 ng/mL ± 278.32 ng/mL, P < 0.05). EA at ST36 significantly preserved ZO-1 protein expression and localization at 12 h after hemorrhage. However, EA at non-channel acupoints had no such effect, and abdominal vagotomy and α-BGT treatment could weaken or eliminate the effects of EA at ST36. Besides, EA at ST36 decreased blood aminotransferase, MB isoenzyme of creatine kinase and creatinine vs EAN group at corresponding time points. At the end of 12-h experiment, the survival rate of the EA group was significantly higher than that of the other groups. CONCLUSION: EA at ST36 attenuates the systemic inflammatory response, protects intestinal barrier integrity, improves organ function and survival rate after hemorrhagic shock via activating the cholinergic anti-inflammatory mechanism.
Assuntos
Eletroacupuntura , Inflamação/terapia , Mucosa Intestinal/metabolismo , Intestinos/inervação , Choque Hemorrágico/terapia , Nervo Vago/fisiopatologia , Animais , Pressão Arterial , Bungarotoxinas/farmacologia , Citocinas/sangue , Modelos Animais de Doenças , Inflamação/sangue , Inflamação/imunologia , Inflamação/patologia , Inflamação/fisiopatologia , Mediadores da Inflamação/sangue , Absorção Intestinal , Intestinos/patologia , Masculino , Permeabilidade , Ratos , Ratos Sprague-Dawley , Choque Hemorrágico/sangue , Choque Hemorrágico/imunologia , Choque Hemorrágico/patologia , Choque Hemorrágico/fisiopatologia , Fatores de Tempo , Vagotomia , Nervo Vago/cirurgia , Proteína da Zônula de Oclusão-1/metabolismoRESUMO
This study investigated the protective effect and mechanism of electroacupuncture at ST36 points on the intestinal barrier dysfunction and remote organ injury after intestinal ischemia and reperfusion injury in rats. Rats were subjected to gut ischemia for 30 min, and then received electroacupuncture for 30 min with or without abdominal vagotomy or intraperitoneal administration of cholinergic α 7 nicotinic acetylcholine receptor ( α 7nAChR) inhibitor. Then we compared its effects with electroacupuncture at nonchannel points, vagal nerve stimulation, or intraperitoneal administration of cholinergic agonist. Cytokine levels in plasma and tissue of intestine, lung, and liver were assessed 60 min after reperfusion. Intestinal barrier injury was detected by histology, gut injury score, the permeability to 4 kDa FITC-dextran, and changes in tight junction protein ZO-1 using immunofluorescence and Western blot. Electroacupuncture significantly lowered the levels of tumor necrosis factor- α and interleukin-8 in plasma and organ tissues, decreased intestinal permeability to FITC-dextran, and prevented changes in ZO-1 protein expression and localization. However, abdominal vagotomy or intraperitoneal administration of cholinergic α 7nAChR inhibitor reversed these effects of electroacupuncture. These findings suggest that electroacupuncture attenuates the systemic inflammatory response through protection of intestinal barrier integrity after intestinal ischemia injury in the presence of an intact vagus nerve.
RESUMO
OBJECTIVE: Burn-induced gut dysfunction plays an important role in the development of sepsis and multiple organ dysfunction. Emerging evidence suggests that hypoxia-inducible factor-1α (HIF-1α) is critical in paracellular barrier functions via regulating vascular endothelial growth factor (VEGF) and myosin light chain kinase (MLCK) expression. Previous studies have also demonstrated that histone deacetylase inhibitors (HDACIs) can repress HIF-1α. This study aims to examine whether valproic acid (VPA), a HDACI, protects against burn-induced gut barrier dysfunction via repressing HIF-1α-dependent upregulation of VEGF and MLCK expression. METHODS: Rats were subjected to third degree 55% TBSA burns and treated with/ without VPA (300 mg/kg). Intestinal barrier dysfunction was evaluated by permeability of intestinal mucosa to fluorescein isothiocyanate (FITC)-dextran and histologic evaluation. Histone acetylation, tight junction protein zonula occludens 1 (ZO-1), VEGF, MLCK and HIF-1α were measured. In addition, CaCO2 cells were transfected with siRNA directed against HIF-1α and were stimulated with CoCl2 (1mM) for 24 hours with/without VPA (2mM) followed by analysis of HIF-1α, MLCK, VEGF and ZO-1. RESULTS: Burn insults resulted in a significant increase in intestinal permeability and mucosal damage, accompanied by a significant reduction in histone acetylation, ZO-1, upregulation of VEGF, MLCK expression, and an increase in HIF-1α accumulation. VPA significantly attenuated the increase in intestinal permeability, mucosa damage, histone deacetylation and changes in ZO-1 expression. VPA also attenuated the increased VEGF, MLCK and HIF-1α protein levels. VPA reduced HIF-1α, MLCK and VEGF production and prevented ZO-1 loss in CoCl2-stimulated Caco-2 cells. Moreover, transfection of siRNA directed against HIF-1α led to inhibition of MLCK and VEGF production, accompanied by upregulation of ZO-1. CONCLUSIONS: These results indicate that VPA can protect against burn-induced gut barrier dysfunction. These protective effects may be due to its inhibitory action on HIF-1α, leading to a reduction in intestinal VEGF and MLCK expression and minimizing ZO-1 degradation.
Assuntos
Queimaduras/complicações , Subunidade alfa do Fator 1 Induzível por Hipóxia/antagonistas & inibidores , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patologia , Ácido Valproico/farmacocinética , Acetilação/efeitos dos fármacos , Animais , Células CACO-2 , Modelos Animais de Doenças , Gastroenterite/tratamento farmacológico , Gastroenterite/etiologia , Gastroenterite/metabolismo , Gastroenterite/patologia , Gastroenterite/prevenção & controle , Histonas/metabolismo , Humanos , Mucosa Intestinal/efeitos dos fármacos , Masculino , Quinase de Cadeia Leve de Miosina/metabolismo , Permeabilidade/efeitos dos fármacos , Substâncias Protetoras/administração & dosagem , Substâncias Protetoras/farmacologia , Ratos , Ácido Valproico/administração & dosagem , Fator A de Crescimento do Endotélio Vascular/metabolismo , Proteína da Zônula de Oclusão-1/metabolismoRESUMO
Objective. Lipid peroxidation plays a critical role in burn-induced plasma leakage, and ulinastatin has been reported to reduce lipid peroxidation in various models. This study aims to examine whether ulinastatin reduces fluid requirements through inhibition of lipid peroxidation in a swine burn model. Methods. Forty miniature swine were subjected to 40% TBSA burns and were randomly allocated to the following four groups: immediate lactated Ringer's resuscitation (ILR), immediate LR containing ulinastatin (ILR/ULI), delayed LR resuscitation (DLR), and delayed LR containing ulinastatin (DLR/ULI). Hemodynamic variables, net fluid accumulation, and plasma thiobarbituric acid reactive substances (TBARS) concentrations were measured. Heart, liver, lung, skeletal muscle, and ileum were harvested at 48 hours after burn for evaluation of TBARS concentrations, activities of antioxidant enzymes, and tissue water content. Results. Ulinastatin significantly reduced pulmonary vascular permeability index (PVPI) and extravascular lung water index (ELWI), net fluid accumulation, and water content of heart, lung, and ileum in both immediate or delayed resuscitation groups. Furthermore, ulinastatin infusion significantly reduced plasma and tissue concentrations of TBARS in both immediate or delayed resuscitation groups. Conclusions. These results indicate that ulinastatin can reduce fluid requirements through inhibition of lipid peroxidation.
Assuntos
Líquidos Corporais/efeitos dos fármacos , Queimaduras/tratamento farmacológico , Glicoproteínas/farmacologia , Glicoproteínas/uso terapêutico , Peroxidação de Lipídeos/efeitos dos fármacos , Animais , Antioxidantes/metabolismo , Pressão Sanguínea/efeitos dos fármacos , Queimaduras/sangue , Queimaduras/enzimologia , Queimaduras/fisiopatologia , Permeabilidade Capilar/efeitos dos fármacos , Modelos Animais de Doenças , Água Extravascular Pulmonar/efeitos dos fármacos , Feminino , Hematócrito , Hemodinâmica/efeitos dos fármacos , Especificidade de Órgãos/efeitos dos fármacos , Sus scrofa , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo , Água/metabolismoRESUMO
OBJECTIVE: To investigate the effects of electroacupuncture at "Zusanli" (ST 36) on the volume of hepatic blood flow, water ratio and plasma alanine aminotransferase (ALT) in rats with delayed fluid replacement after hemorrhagic shock and to provide the references for electroacupuncture at Zusanli (ST 36) in treating hemorrhagic shock. METHODS: Forty SD rats with hemorrhagic shock induced by bloodletting 40% of whole blood volume were randomly divided into a hemorrhage with no treatment (NT) group, an immediate fluid replacement (IFR) group, an electroacupuncture at Zusanli (ST 36) and delayed fluid resuscitation (EA/DFR) group and a sham electroacupuncture and delayed fluid replacement (SEA/DFR) group, 10 rats in each group. No treatment was performed in NT group. IFR group was treated with fluid replacement at 10 minutes after blood loss, and EA/DFR group was treated with electroacupuncture at "Zusanli" (ST 36) at 10 minutes after blood loss, while non-acupoint was punctured in SEA/DFR group. Two EA groups were received delayed fluid replacement at 3 hours after blood loss. The volume of hepatic blood flow and ALT before blood loss and 3 h and 12 h after blood loss, and water ratio 12 h after blood loss were measured. RESULTS: After blood loss, all parameters in IFR group and EA/DFR group were improved significantly in contrast with those in NT group (all P < 0.05). There was no significant difference between SEA/DFR group and NT group. Three hours after blood loss, the hepatic blood flow of IFR group was significant higher than those of NT group, EA/DFR group and SEA/DFR group (all P < 0.05), while the plasma ALT of IFR group was significant lower than those of NT group, EA/DFR group and SEA/DFR group (all P < 0.05), and the plasma ALT of EA/DFR group was lower than those of NT group and SEA/DFR group (both P < 0.05), the hepatic blood flow of EA/DFR group showed no significant difference compared with that of SEA/DFR group (P > 0.05). Twelve hours after blood loss, the plasma ALT and the water ratio of EA/DFR group and IFR group were significant lower than those of NT group and SEA/DFR group (all P < 0.05), and the hepatic blood flow of EA/DFR group and IFR group was significant higher than those of NT group and SEA/DFR group (all P < 0.05), while the plasma ALT of IFR group was significant lower than that of EA/DFR group (P < 0.05), and the hepatic blood flow of IFR group was significant higher than that of EA/DFR group (P < 0.05). CONCLUSION: Electroacupuncture at "Zusanli" (ST 36) has a protective effects for hepatic ischemic injury in rats with delayed fluid replacement after hemorrhagic shock.
Assuntos
Pontos de Acupuntura , Eletroacupuntura , Isquemia/terapia , Fígado/irrigação sanguínea , Choque Hemorrágico/terapia , Animais , Humanos , Masculino , Ratos , Ratos Sprague-DawleyRESUMO
Diffuse pulmonary lymphangiomatosis (DPL) is a rare disease that is characterized by diffuse proliferation of abnormal pulmonary lymphatic channels. DPL occurs mostly in children and young adults and often undergoes a progressive clinical course, eventually causing deterioration of the lung. Both the clinical diagnosis and treatment of DPL remain a challenge. Here, we report a case of DPL in a 53-year-old Chinese woman with comprehensive investigations including pulmonary function tests, computer tomography (CT), bronchoscopy and histological examination of the lung biopsy, and review the literature.