RESUMO
OBJECTIVE: Diabetic nephropathy (DN) is a serious chronic complication of diabetes mellitus (DM). Endoplasmic reticulum (ER) stress is an important factor in the regulation of pathological processes in DN, and excessive ER stress can lead to apoptosis. Although the IL-33/ST2 axis is known to be involved in diabetic kidney disease or related nephropathy, its role and molecular mechanisms remain poorly understood in terms of DN. The purpose of this study was to investigate the effects of IL-33/ST2 signaling on DN and to characterize the roles that ER stress and apoptosis play in DN. METHODS: To investigate this study, mice were randomly assigned into DN (induced by 0.1% STZ) and Control groups. Biochemical indices (FBG, BUN, UPR, UCE) were measured in serum and urine samples to reflect blood glucose and kidney damage. Quantitative real-time PCR, western blot, and immunofluorescence were used to assess gene and protein expression of the IL-33/ST2 axis and ER stress relative signaling molecule. Apoptosis was analyzed by flow cytometry. RESULTS: IL-33 levels are significantly increased in the kidneys of patients and mice with DN. Double immunofluorescence staining showed that IL-33 colocalized with CD31-positive endothelial cells. Treatment with IL-33 attenuated kidney injury in Streptozotocin (STZ)-treated mice. In vitro, we showed that IL-33 attenuated ER stress and apoptosis in glomerular endothelial cells. However, sST2 treatment significantly reversed these effects of IL-33. CONCLUSION: Together, these data suggest that IL-33/ST2 signaling mitigates STZ-induced renal damage, partly at least, by suppressing ER stress and apoptosis. Therefore, IL-33 may be an effective therapeutic target in DN.
Assuntos
Diabetes Mellitus Experimental , Nefropatias Diabéticas , Ratos , Humanos , Camundongos , Animais , Nefropatias Diabéticas/patologia , Células Endoteliais/metabolismo , Interleucina-33/farmacologia , Interleucina-33/uso terapêutico , Proteína 1 Semelhante a Receptor de Interleucina-1 , Ratos Sprague-Dawley , Diabetes Mellitus Experimental/metabolismo , Estresse do Retículo Endoplasmático , ApoptoseRESUMO
We had previously demonstrated that the calcitonin gene-related peptide (CGRP) suppresses the oxidative stress and vascular smooth muscle cell (VSMC) proliferation induced by vascular injury. A recent study also indicated that CGRP protects against the onset and development of angiotensin II (Ang II)-induced hypertension, vascular hypertrophy and oxidative stress. However, the mechanism behind the effects of CGRP on Ang II-induced oxidative stress is unclear. CGRP significantly suppressed the level of reactive oxygen species (ROS) generated by NADPH oxidase in Ang II-induced VSMCs. The Ang II-stimulated activation of both Src and the downstream transcription factor, STAT3, was abrogated by CGRP. However, the antioxidative effect of CGRP was lost following the expression of constitutively activated Src or STAT3. Pre-treatment with H-89 or CGRP8-37 also blocked the CGRP inhibitory effects against Ang II-induced oxidative stress. Additionally, both in vitro and in vivo analyses show that CGRP treatment inhibited Ang II-induced VSMC proliferation and hypertrophy, accompanied by a reduction in ROS generation. Collectively, these results demonstrate that CGRP exhibits its antioxidative effect by blocking the Src/STAT3 signalling pathway that is associated with Ang II-induced VSMC hypertrophy and hyperplasia.
Assuntos
Angiotensina II/farmacologia , Peptídeo Relacionado com Gene de Calcitonina/metabolismo , NADPH Oxidases/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Fator de Transcrição STAT3/metabolismo , Transdução de Sinais/fisiologia , Quinases da Família src/metabolismo , Animais , Antioxidantes/metabolismo , Calcitonina/metabolismo , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Músculo Liso Vascular/metabolismo , Miócitos de Músculo Liso/metabolismo , Oxirredução/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/fisiologia , Ratos , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacosRESUMO
Apoptosis is associated with various cardiovascular diseases. CGRP exerts a variety of effects within the cardiovascular system, and protects against the onset and development of angiotensin (Ang) II-induced vascular dysfunction and remodelling. However, it is not known whether CGRP has a direct effect on Ang II-induced apoptosis in vascular smooth muscle cells (VSMCs), and the mechanism underlying the anti-apoptotic role remains unclear. In this study, CGRP significantly suppressed reactive oxygen species (ROS) and apoptosis in Ang II-induced VSMCs. In VSMCs pre-treated with a CGRP receptor antagonist (CGRP8-37), the CGRP-mediated inhibition of Ang II-induced ROS and apoptosis was completely abolished. Moreover, pre-treatment with N-acetyl-L cysteine (NAC), an ROS scavenger, blocked the effects of CGRP on Ang II-induced apoptosis. In addition, the activation of CaMKII and the downstream transcription factor CREB stimulated by Ang II was abrogated by CGRP. Importantly, in both CGRP and NAC-treated VSMCs, CGRP failed to further attenuate CaMKII and CREB activation. The results demonstrate that CGRP attenuated Ang II-induced ROS-dependent apoptosis in VSMCs by inhibiting the CaMKII/CREB signalling pathway.
Assuntos
Angiotensina II/farmacologia , Apoptose , Peptídeo Relacionado com Gene de Calcitonina/fisiologia , Músculo Liso Vascular/citologia , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais/efeitos dos fármacos , Animais , Proteína Quinase Tipo 2 Dependente de Cálcio-Calmodulina/antagonistas & inibidores , Proteína Quinase Tipo 2 Dependente de Cálcio-Calmodulina/metabolismo , Células Cultivadas , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , HumanosRESUMO
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.
Assuntos
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
BACKGROUND: Recent findings showed advantages of a novel pyruvate-enriched oral rehydration solution (Pyr-ORS) in resuscitation of burns. This study focused on effects of Pyr-ORS on the visceral blood perfusion (VBP), gastrointestinal function, and survival rate, compared with the bicarbonate-based World Health Organization-guided oral rehydration solution (WHO-ORS), during intragastric rehydration of lethal hemorrhagic shock in rats. METHODS: Sixty adult rats were subjected to 45% total blood volume loss and were randomly allocated to the following three groups (n = 20): group NR (no fluid resuscitation), group PORS (oral Pyr-ORS rehydration), and group BORS (oral WHO-ORS rehydration), respectively. Other 10 rats were served as group NH (the sham group). Enteral rehydration lasted for 4 h after hemorrhage. The mean arterial pressure (MAP), VBP, and plasma enzymes activities of heart, liver, and kidney, and intestinal fatty acid binding protein were measured. Liver, kidney, and ileum were harvested for the evaluation of activities of oxidative enzymes and intestinal barrier protein (ZO-1). Other 84 rats with identical procedures without sampling were observed for their 24-h survival rates. RESULTS: Pyr-ORS was more effective in enhancing the MAP and VBP, inhibiting tissue oxidative damage, and improving organ function, compared with WHO-ORS. Hypoxic lactic acidosis was fully corrected in group PORS in 4 h, whereas it worsened in group BORS, and the 24-h survival rate was twice higher in group PORS than in group BORS (45.8 versus 20.8%, P < 0.05). CONCLUSIONS: A small amount of pyruvate in Pyr-ORS was more therapeutically beneficial than equivalent bicarbonate in WHO-ORS and greatly raised survival in enteral rehydration of lethal hemorrhagic shock. The Pyr-ORS may be an ideal oral fluid in resuscitation of hypovolemic shock, especially in prehospital and resource-poor settings.
Assuntos
Hidratação/métodos , Ácido Pirúvico/farmacologia , Ressuscitação/métodos , Choque Hemorrágico/tratamento farmacológico , Acidose Láctica/tratamento farmacológico , Acidose Láctica/metabolismo , Animais , Bicarbonatos/farmacologia , Modelos Animais de Doenças , Glucose/farmacologia , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Estresse Oxidativo/efeitos dos fármacos , Cloreto de Potássio/farmacologia , Distribuição Aleatória , Ratos Sprague-Dawley , Choque Hemorrágico/metabolismo , Cloreto de Sódio/farmacologia , Taxa de Sobrevida , Resultado do Tratamento , Vísceras/irrigação sanguíneaRESUMO
Clodronate liposome injection is an effective approach to selectively and specifically depleting macrophages. Macrophages play a crucial role in cutaneous wound healing and are associated with excessive scar formation. Use of clodronate liposomes to enhance cutaneous wound healing and reduce scar formation could represent a major advance in wound therapy and hypertrophic scar treatment. This study aimed to investigate the effects of subcutaneous or intraperitoneal injection of clodronate liposomes on cutaneous wound healing and scar formation. A burn injury mouse model was used. Mice were treated with subcutaneous or intraperitoneal injection of clodronate liposomes. Wound healing time was analyzed and scar tissues were harvested for hematoxylin and eosin (HE) staining, reverse transcription polymerase chain reaction (RT-PCR) and Western blot analyses. Wound healing time in treated mice was extended. HE showed that the basal layer of the epidermis in treated scars was flattened, the dermis layer was not significantly thickened, and collagen fibers were well arranged, with few cells and micro vessels. RT-PCR and Western blot analyses showed that the levels of TGF-ß1 and collagen I-α2 were decreased in treated mice. Clodronate liposomes reduce excessive scar formation and delay cutaneous wound healing possibly by reducing collagen deposition and macrophage-derived TGF-ß1 expression.
Assuntos
Queimaduras/metabolismo , Queimaduras/patologia , Cicatriz/metabolismo , Ácido Clodrônico/administração & dosagem , Colágeno/metabolismo , Fator de Crescimento Transformador beta1/metabolismo , Animais , Queimaduras/tratamento farmacológico , Cicatriz/tratamento farmacológico , Cicatriz/patologia , Colágeno Tipo I/metabolismo , Modelos Animais de Doenças , Lipossomos , Macrófagos/imunologia , Macrófagos/patologia , Camundongos , Fatores de Tempo , Cicatrização/efeitos dos fármacos , Cicatrização/imunologiaRESUMO
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
The aim of this study was to examine whether administration of valproic acid (VPA), a histone deacetylase inhibitor, inhibits proinflammatory mediators and ameliorate visceral vasopermeability both in a rat model of major burn, and also in rat cultured endothelial cells stimulated with permeability evoking mediators. SD rats were subjected to a 50% TBSA full-thickness scald injury, and treated with either saline or VPA (300 mg/kg) intraperitoneally. Pulmonary vascular endothelial growth factor (VEGF), myeloperoxidase (MPO), pulmonary microvascular permeability, water content, and acetylation of histone H3K9 of lungs were evaluated. In addition, pulmonary microvascular endothelial cells (PMECs) from male SD rats were cultured. With then, MPO, VEGF, histone acetylation, and the permeability of PMECs were investigated. Lethal scald injury resulted in a significant increase in microvascular permeability and water content of lung, accompanied by a significant elevation of the content of VEGF and activity of MPO, and a decrease of histone acetylation. VPA treatment significantly alleviated the microvascular permeability and water content of lung, lowered the levels of VEGF and MPO, and promoted acetylation of histone H3K9 following scald injury. Moreover, VPA reduced permeability of monolayer PMECs subjected to scald serum challenge, reduced the level of MPO and VEGF in supernatants, and promoted acetylation of histone H3K9 in PMECs. These results indicated that VPA can protect pulmonary microvascular endothelial barrier, alleviate proinflammatory mediators-evoked vascular hyperpermeability and tissue edema and improve the survival rate of rats subjected to lethal scald injury.
Assuntos
Queimaduras/tratamento farmacológico , Queimaduras/metabolismo , Permeabilidade Capilar/efeitos dos fármacos , Células Endoteliais/efeitos dos fármacos , Inibidores Enzimáticos/uso terapêutico , Ácido Valproico/uso terapêutico , Animais , Técnicas de Cultura de Células , Modelos Animais de Doenças , Histona Acetiltransferases/metabolismo , Masculino , Peroxidase/metabolismo , Ratos , Ratos Sprague-Dawley , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
Inflammation is a defensive response in the living tissue of the vascular system that acts against damage factors and involves various types of immune cells, including macrophages, neutrophils, endothelial cells and other associated immune molecules. If the release of inflammatory mediators is excessive, systemic inflammatory response syndrome may develop. Sepsis is the most common complication of severe burns and is a systemic inflammatory response syndrome that is caused by infectious factors and is capable of leading to multiple organ dysfunction and potentially death. Research concerning the mechanism and treatment of sepsis is crucial. Macrophages are an important type of immune cell that remove invasive pathogens and are involved in innate and adaptive immune responses. It has been previously reported that bone marrow mesenchymal stem cells (BMSCs) affect macrophages by regulating immunity. The present study aimed to investigate the effect of BMSCs on macrophage polarization in vivo and in vitro, in addition to the potential therapeutic effect of these cells on experimental sepsis. BMSCs and peritoneal macrophages were isolated from SpragueDawley rats and cocultured overnight as a mixed culture or Transwell system, and subsequently stimulated with 100 ng/ml lipopolysaccharide (LPS). After 12 h, the medium was replaced with normal complete medium for various durations and supernatants were collected to extract proteins and cells for ELISA, western blot and flow cytometry analysis to investigate different aspects of macrophages. Sepsis was induced in SpragueDawley rats by injection of LPS (5 mg/kg), followed by tail vein injection of BMSCs or PBS 1 h later. After 6, 12, 24 and 48 h, lung tissues were harvested for pathological observation and peritoneal macrophages were collected for flow cytometry analysis to assess the expression of markers, including cluster of differentiation (CD)68 (used for gating), CD11c and CD206. The results demonstrated that, in the culture medium, LPS stimulation increased the expression of CD11c in macrophages, and the levels of tumor necrosis factorα and inducible nitric oxide synthase were also increased. By contrast, in macrophages treated with BMSCs directly, the expression of CD11c was reduced compared with the LPSstimulated macrophage alone group. However, the secretion of interleukin10, transforming growth factorß and arginase1 was increased in the direct coculture group, compared with the LPSstimulated macrophage alone group. BMSCs reduced the inflammation in lung tissues and inhibited macrophage expression of CD11c in the rat model of sepsis. The results of the present study demonstrated that BMSCs cocultured with macrophages directly inhibited macrophage differentiation into the M1 phenotype and reduced inflammation in macrophages stimulated by LPS. In vivo, BMSCs decreased the expression of CD11c in peritoneal macrophages and reduced the pathological inflammatory response in the lungs. The findings of the present study demonstrated that BMSCs may reduce the extent of the systemic inflammatory response, which may contribute to the development for a novel type of treatment for sepsis in the future.
Assuntos
Macrófagos Peritoneais/citologia , Células-Tronco Mesenquimais/citologia , Animais , Antígenos CD/metabolismo , Antígenos de Diferenciação Mielomonocítica/metabolismo , Arginase/metabolismo , Células da Medula Óssea/citologia , Antígeno CD11c/metabolismo , Diferenciação Celular , Polaridade Celular/efeitos dos fármacos , Células Cultivadas , Técnicas de Cocultura , Citocinas/metabolismo , Modelos Animais de Doenças , Lipopolissacarídeos/farmacologia , Pulmão/patologia , Macrófagos Peritoneais/imunologia , Macrófagos Peritoneais/metabolismo , Masculino , Células-Tronco Mesenquimais/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Ratos , Ratos Sprague-Dawley , Sepse/metabolismo , Sepse/patologiaRESUMO
OBJECTIVE: To explore the effect of dimethyl sulfoxide(DMSO) on suppressing the release of gut inflammatory cytokine and re-store of the barrier impairment following zymosan-insulted systemic inflammatory response syndrome (SIRS). METHODS: S D rats were randomly divided into four groups:sham with administration of normal saline (SS group); sham with administration of DMSO (DS group); zymosan with administration of normal saline (ZS group); and zymosan with administration of DMSO (ZD group), each group includes two subgroups at 4 h and 24 h after surgery. At 4 h and 24 h after intraperitoneal injection of zymosan (750 mg/kg), the levels of intestinal inflammatory cytokines (tumor necrosis factor-αand interleukin-10) and the activity of diamine oxidase in plasma were examined. Intestinal injury was evaluated by using an intestinal histological score. RESULTS: DMSO suppressed the release of tumor necrosis factor-αand increased interleukin-10 levels in the intestine compared with the ZS group at the corresponding time points. DMSO decreased the level of diamine oxidase in plasma compared with the ZS group. DMSO restored the injury of intestinal villi and the gut injury score was significantly lower than that in the ZS group. CONCLUSIONS: DMSO can suppress the release of intestinal inflammatory cytokines and restore zymosan-insulted gut barrier impairment.
Assuntos
Dimetil Sulfóxido/farmacologia , Mucosa Intestinal/efeitos dos fármacos , Zimosan/efeitos adversos , Amina Oxidase (contendo Cobre)/sangue , Animais , Citocinas/metabolismo , Inflamação , Substâncias Protetoras/farmacologia , Ratos , Ratos Sprague-DawleyRESUMO
BACKGROUND: Recent studies have suggested that pyruvate-enriched oral rehydration solution (Pyr-ORS) may be superior to the standard bicarbonate-based ORS in the protection of intestine from ischemic injury. The aim of this study was to compare the effects of Pyr-ORS with citrate-enriched ORS (Cit-ORS) on the intestinal hypoxia-inducible factor-1 (HIF-1)-erythropoietin (EPO) signaling pathway for enteral rehydration in a rat model of burn injury. METHODS: Rats were randomly assigned to 4 groups (N = 20, 2 subgroups each: n = 10): scald sham (group SS), scald with no fluid resuscitation (group SN), scald and resuscitation with enteral Cit-ORS (group SC), and scald and resuscitation with enteral Pyr-ORS (group SP). At 2.5 and 4.5 hours after a 35% total body surface area (TBSA) scald, intestinal mucosal blood flow (IMBF), contents of HIF-1, EPO, endothelial nitric oxide synthase (eNOS), nitric oxide (NO), barrier protein (ZO-1), levels of serum diamine oxidase (DAO), and intestinal mucosal histology injury score were determined. RESULTS: Serum DAO activities in the scalded groups were significantly elevated, but less raised in group SP than in group SC, at 2.5 hours and at 4.5 hours after the scald. Further, group SP more profoundly preserved intestinal HIF-1 expression compared with group SC at the 2 time points. Compared with group SC, group SP had markedly elevated intestinal EPO, eNOS, and NO levels at the same time points, respectively (P < .05). Similarly, IMBF and ZO-1 levels were significantly higher in group SP than in group SC. Intestinal mucosal histopathological scores were statistically higher at 2.5 hours and 4.5 hours after scalding but were more attenuated in group SP than in group SC (P < .05). Immunofluorescence expression of intestinal mucosal ZO-1 was consistent with the above changes. The above parameters were also significantly different between groups SC and SN (all P < .05). CONCLUSION: Pyr-ORS provides a superior option to Cit-ORS for the preservation of intestinal blood flow and barrier function and the attenuation of histopathological alterations in enteral resuscitation of rats with burn injury. Its underlying mechanism may be closely related to the pyruvate in activation of intestinal HIF-1-EPO signaling cascades.
Assuntos
Queimaduras/tratamento farmacológico , Ácido Cítrico/administração & dosagem , Fator 1 Induzível por Hipóxia/metabolismo , Intestinos/efeitos dos fármacos , Ácido Pirúvico/administração & dosagem , Amina Oxidase (contendo Cobre)/sangue , Amina Oxidase (contendo Cobre)/genética , Animais , Bicarbonatos/química , Superfície Corporal , Eritropoetina/genética , Eritropoetina/metabolismo , Hidratação , Glucose/química , Fator 1 Induzível por Hipóxia/genética , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/metabolismo , Masculino , Óxido Nítrico Sintase Tipo III/genética , Óxido Nítrico Sintase Tipo III/metabolismo , Cloreto de Potássio/química , Ratos , Ratos Sprague-Dawley , Ressuscitação , Transdução de Sinais , Cloreto de Sódio/química , Proteína da Zônula de Oclusão-1/genética , Proteína da Zônula de Oclusão-1/metabolismoRESUMO
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.
Assuntos
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
BACKGROUND: Several epidemiological studies have evaluated the association between the NAD(P)H oxidase p22 phox gene C242T polymorphism and the risk of type 2 diabetes mellitus (T2DM), diabetic nephropathy (DN), and carotid atherosclerosis with T2DM (CA), but the results are inconclusive. This meta-analysis was therefore designed to clarify these controversies. METHODS: Systematic searches were performed using electronic databases such as MEDLINE, PubMed, EMBASE, and China National Knowledge Infrastructure, as well as through manual searching of the references of identified articles. A total of 11 publications were eligible for this meta-analysis after running a search on the NAD(P)H oxidase p22 phox gene C242T polymorphism, including 7 with outcomes for T2DM, 7 with outcomes for DN, and 3 with outcomes for CA. The pooled odds ratio (OR) with a 95% confidence interval (CI) was calculated using a fixed effects model (FEM) or a random effects model (REM). Publication bias was tested by Begg's funnel plot analysis. Sensitivity analysis was also performed. RESULTS: The results showed a significant association between the NAD(P)H oxidase p22 phox gene C242T polymorphism and T2DM risk in the allelic model (REM: OR = 1.23, 95% CI = 1.06-1.43), additive model (FEM: OR = 1.61, 95% CI = 1.14-2.26), and recessive model (FEM: OR = 1.50, 95% CI = 1.10-2.05). A significant association was also observed for DN in the allelic model (REM: OR = 1.25, 95% CI = 1.06-1.47), additive model (FEM: OR = 1.61, 95% CI = 1.08-2.38), and dominant model (REM: OR = 1.26, 95% CI = 1.03-1.54). However, no association was observed for CA. Similar results were obtained in subgroup analysis based on ethnicity. CONCLUSIONS: Results of this meta-analysis suggest that the NAD(P)H oxidase p22 phox gene 242T allele might be associated with an increased risk of T2DM and DN, but not CA.
RESUMO
AIMS: The objective of this study was to investigate the effects of pyruvate-containing fluids on peritoneal resuscitation (PR), following intravenous fluid resuscitation from hemorrhagic shock (HS) in rats. METHODS: One hundred rats following 1-h HS with mean arterial pressure 35 ± 5 mmHg were randomly assigned to five groups (n = 10) in each of two comparable sets: group VR: intravenous resuscitation (VR) only and four groups with PR after VR: groups NS, LA, P1, and P2, resuscitated with normal saline, lactated peritoneal dialysis solution (PDS), pyruvated PDS, and 2.2% pyruvate, respectively. The splanchnic blood flow on surfaces of liver, kidney, and intestinal mucosa was detected. Blood samples were taken before HS and at T180 or T360 in these two animal sets after hemorrhage for function tests of liver, kidney, and intestinal mucosa, respectively. The intestinal mucosal barrier protein: zonula occludens 1 (ZO-1) and tissue water contents of these organs were also determined. RESULTS: Splanchnic blood flow was significantly preserved in all PR groups with hyperosmolar solutions: group P1 and group P2 with pyruvate were more advantageous than group LA. Group P2 was the most efficient among groups in reverse of visceral hypoperfusion. Organ function and tissue water contents of liver, kidney, and intestine and the intestinal barrier ZO-1 density were also improved in group P1 and group P2, compared with group LA. Among organs, the pyruvate protection of intestinal mucosa was the most apparent by reversing splanchnic blood flow and diamine oxidase close to reference ranges with the highest ZO-1 density. Group P2 showed the most pyruvate protection in all test parameters among four groups with PR. CONCLUSIONS: Peritoneal resuscitation with hyperosmolar fluids attenuated visceral vasoconstriction and splanchnic hypoperfusion and improved the intestinal barrier protein and organ function following conventional fluid resuscitation from severe HS in rats. Pyruvate was superior to lactate in PDS as PR fluids, and 2.2% pyruvate was the optimal fluid in PR.
Assuntos
Hidratação/métodos , Ácido Pirúvico/uso terapêutico , Soluções para Reidratação/uso terapêutico , Ressuscitação/métodos , Choque Hemorrágico/terapia , Animais , Biomarcadores/sangue , Pressão Sanguínea/efeitos dos fármacos , Água Corporal/metabolismo , Infusões Parenterais , Mucosa Intestinal/irrigação sanguínea , Rim/irrigação sanguínea , Fígado/irrigação sanguínea , Masculino , Ácido Pirúvico/farmacologia , Ratos , Ratos Sprague-Dawley , Fluxo Sanguíneo Regional/efeitos dos fármacos , Soluções para Reidratação/farmacologia , Choque Hemorrágico/metabolismo , Choque Hemorrágico/fisiopatologia , Circulação Esplâncnica/efeitos dos fármacos , Proteína da Zônula de Oclusão-1/metabolismoRESUMO
AIM: To investigate alteration in intestinal absorption during enteral resuscitation with pyruvate-enriched oral rehydration solution (Pyr-ORS) in scalded rats. METHODS: To compare pyruvate-enriched oral rehydration solution (Pyr-ORS) with World Health Organisation oral rehydration solution (WHO-ORS), 120 rats were randomly divided into 6 groups and 2 subgroups. At 1.5 and 4.5 h after a 35% TBSA scald, the intestinal absorption rate, mucosal blood flow (IMBF), Na(+)-K(+)-ATPase activity and aquaporin-1 (AQP-1) expression were determined (n = 10), respectively. RESULTS: The intestinal Na(+)-K(+)-ATPase activity, AQP-1 expression and IMBF were markedly decreased in scald groups, but they were profoundly preserved by enteral resuscitation with WHO-ORS and further improved significantly with Pyr-ORS at both time points. Na(+)-K+-ATPase activities remained higher in enteral resuscitation with Pyr-ORS (Group SP) than those with WHO-ORS (Group SW) at 4.5 h. AQP-1 and IMBF were significantly greater in Group SP than in Group SW at both time points. Intestinal absorption rates of water and sodium were obviously inhibited in scald groups; however, rates were also significantly preserved in Group SP than in Group SW with an over 20% increment at both time points. CONCLUSION: The Pyr-ORS may be superior to the standard WHO-ORS in the promotion of intestinal absorption of water and sodium during enteral resuscitation.
Assuntos
Aquaporina 1/efeitos dos fármacos , Queimaduras , Absorção Intestinal/efeitos dos fármacos , Mucosa Intestinal/efeitos dos fármacos , Ácido Pirúvico/farmacologia , Soluções para Reidratação/farmacologia , ATPase Trocadora de Sódio-Potássio/efeitos dos fármacos , Sódio/metabolismo , Água/metabolismo , Animais , Aquaporina 1/metabolismo , Bicarbonatos/farmacologia , Hidratação , Glucose/farmacologia , Mucosa Intestinal/irrigação sanguínea , Mucosa Intestinal/metabolismo , Fluxometria por Laser-Doppler , Masculino , Cloreto de Potássio/farmacologia , Ratos , Ratos Sprague-Dawley , Cloreto de Sódio/farmacologia , ATPase Trocadora de Sódio-Potássio/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.
Assuntos
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.
Assuntos
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
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
BACKGROUND: The aim of this study was to examine whether administration of ulinastatin inhibits pro-inflammatory mediators and ameliorate visceral vasopermeability both in a rat model of major burn, and also in rat cultured endothelial cells stimulated with permeability-evoking mediators. METHODS: Plasma levels of tumor necrosis factor-alpha (TNF-α), C-reactive protein (CRP), myeloperoxidase (MPO), microvascular permeability, and water content of organ tissues were evaluated in a rodent model of a 55% TBSA full-thickness scald injury. Microvascular permeability was also evaluated with a cultured pulmonary microvascular endothelial cells (PMECs) monolayer after stimulation with trypsin, bradykinin, histamine, prostaglandin E2 and burn serum. RESULTS: We found that the plasma levels of TNF-α, CRP, MPO, vascular permeability and water content of heart, lung, kidney, and small intestine tissues were significantly increased in animals after scald injury, and administration of ulinastatin lowered the levels TNF-α, CRP, MPO, vascular permeability and water content of those organ tissues. In vitro, ulinastatin lowered the levels of TNF-α, interleukin-6 (IL-6) and attenuated permeability in PMEC monolayers after being stimulated with burn serum or trypsin, but not by bradykinin, histamine or prostaglandin E2. CONCLUSIONS: These results indicate that ulinastatin attenuates the systemic inflammatory response and visceral vasopermeability both in vivo and vitro, and may serve as a therapeutic agent for prevention of systemic inflammatory response and leakage of fluid into tissue after major burn.
Assuntos
Queimaduras/tratamento farmacológico , Permeabilidade Capilar/efeitos dos fármacos , Glicoproteínas/farmacologia , Mediadores da Inflamação/metabolismo , Inibidores da Tripsina/farmacologia , Água/metabolismo , Animais , Biomarcadores/metabolismo , Queimaduras/metabolismo , Proteína C-Reativa/análise , Modelos Animais de Doenças , Inflamação/metabolismo , Interleucina-6/metabolismo , Intestino Delgado/metabolismo , Rim/metabolismo , Pulmão/metabolismo , Masculino , Peroxidase/metabolismo , Ratos , Fator de Necrose Tumoral alfa/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.