RESUMO
BACKGROUND: Mitochondrial transcription factor A (TFAM) plays multiple pathophysiologic roles in mitochondrial DNA (mtDNA) maintenance. However, the role of TFAM in sepsis-induced acute kidney injury (AKI) remains largely unknown. METHODS: Lipopolysaccharide (LPS) treatment of HK-2 cells mimics the in vitro model of AKI inflammation. pcDNA3.1 plasmid was used to construct pcDNA3.1-TFAM. sh-TFAM-543, sh-TFAM-717, sh-TFAM-765, sh-TFAM-904 and pcDNA3.1-TFAM were transfected into HK-2 cells using Lipofectamine 2000. MtDNA transcriptional levels were detected by quantitative real-time polymerase chain reaction (qRT-PCR). 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide (MTT) assay was performed to assess the cell viability. Changes in reactive oxygen species (ROS) and mitochondrial membrane potential in HK-2 cells were detected using the corresponding kits. Immunofluorescence experiment was used to investigate the displacement of TFAM. mRNA and protein expression levels of TFAM and its related genes were measured by qRT-PCR and western blot respectively. Mice in sepsis were administered cecal ligation and puncture surgery. RESULTS: LPS treatment was a non-lethal influencing factor, leading to the upregulation of ROS levels and downregulation of mtDNA copy number and NADH dehydrogenase subunit-1 (ND1) expression, and caused damage to the mitochondria. As the LPS treatment time increased, TFAM was displaced from the periphery of the nucleus to cytoplasm. TFAM reduced ROS and P38MAPK levels by inhibiting toll-like receptor 4 (TLR4) expression, ultimately inhibiting inflammation and repairing mtDNA. CONCLUSIONS: Our results indicate that TFAM repairs mtDNA by blocking the TLR4/ROS/P38MAPK signaling pathway in inflammatory cells, thereby repairing septic tubular epithelial cells, and TFAM may serve as a new target for sepsis therapy.
Assuntos
Proteínas de Ligação a DNA/genética , Células Epiteliais/patologia , Proteínas Mitocondriais/genética , Espécies Reativas de Oxigênio/metabolismo , Sepse/genética , Transdução de Sinais/efeitos dos fármacos , Receptor 4 Toll-Like/antagonistas & inibidores , Fatores de Transcrição/genética , Proteínas Quinases p38 Ativadas por Mitógeno/antagonistas & inibidores , Animais , Linhagem Celular , Humanos , Túbulos Renais/citologia , Camundongos , Camundongos Endogâmicos C57BL , Sepse/patologiaRESUMO
BACKGROUND: Sepsis and sepsis-associated encephalopathy (SAE) are common intensive care unit (ICU) diseases; the morbidity and mortality are high. The present study analyzed the sensitivity of different diagnostic criteria of sepsis 1.0 and 3.0, epidemiological characteristics of sepsis and SAE, and explored its risk factors for death, short-term, and long-term prognosis. METHODS: The retrospective study included patients in ICU from January 2015 to June 2016. After excluding 58 patients, 175 were assigned to either an SAE or a non-SAE group (patients with sepsis but no encephalopathy). The sensitivity of the diagnostic criteria was compared between sepsis 1.0 and 3.0, respectively. Between-group differences in baseline data, Acute Physiology and Chronic Health Evaluation II score (APACHE II score), Sequential Organ Failure Assessment score (SOFA score), etiological data, biochemical indicators, and 28-day and 180-day mortality rates were analyzed. Survival outcomes and long-term prognosis were observed, and risk factors for death were analyzed through 180-day follow-up. RESULTS: The sensitivity did not differ significantly between the diagnostic criteria of sepsis 1.0 and 3.0 (P = .286). The 42.3% incidence of SAE presented a significantly high APACHE II and SOFA scores as well as 28-day mortality and 180-day mortality (all P < .001). The incidence of death was 37.1%. The multivariate stepwise regression analysis demonstrated that the risk of death in SAE group was significantly higher than the non-SAE group (P < .001). Sepsis-associated encephalopathy is a risk factor for sepsis-related death (relative risk [RR] = 2.868; 95% confidence interval: 1.730-4.754; P < .001). Although males showed a significantly high rate of 28-day and 180-day mortality (P = .035 and .045), it was not an independent risk factor for sepsis-related death (P = .072). The long-term prognosis of patients with sepsis was poor with decreased quality of life. No significant difference was observed in prognosis between the SAE and non-SAE groups (P > .05). CONCLUSION: Both diagnostic criteria cause misdiagnosis, and the sensitivity did not differ significantly. The incidence of SAE was high, and 28-day and 180-day mortality rates were significantly higher than those without SAE. Sepsis-associated encephalopathy is a risk factor for poor outcome. The overall long-term prognosis of patients with sepsis was poor, and the quality of life decreased.
Assuntos
APACHE , Escores de Disfunção Orgânica , Encefalopatia Associada a Sepse/mortalidade , Sepse/mortalidade , Adulto , Idoso , Feminino , Mortalidade Hospitalar , Humanos , Incidência , Unidades de Terapia Intensiva/estatística & dados numéricos , Masculino , Pessoa de Meia-Idade , Prognóstico , Qualidade de Vida , Estudos Retrospectivos , Fatores de Risco , Sepse/patologia , Encefalopatia Associada a Sepse/patologiaRESUMO
BACKGROUND: Poor inter-rater reliability in chest radiograph interpretation has been reported in the context of acute respiratory distress syndrome (ARDS), although not for the Berlin definition of ARDS. We sought to examine the effect of training material on the accuracy and consistency of intensivists' chest radiograph interpretations for ARDS diagnosis. METHODS: We conducted a rater agreement study in which 286 intensivists (residents 41.3%, junior attending physicians 35.3%, and senior attending physician 23.4%) independently reviewed the same 12 chest radiographs developed by the ARDS Definition Task Force ("the panel") before and after training. Radiographic diagnoses by the panel were classified into the consistent (n = 4), equivocal (n = 4), and inconsistent (n = 4) categories and were used as a reference. The 1.5-hour training course attended by all 286 intensivists included introduction of the diagnostic rationale, and a subsequent in-depth discussion to reach consensus for all 12 radiographs. RESULTS: Overall diagnostic accuracy, which was defined as the percentage of chest radiographs that were interpreted correctly, improved but remained poor after training (42.0 ± 14.8% before training vs. 55.3 ± 23.4% after training, p < 0.001). Diagnostic sensitivity and specificity improved after training for all diagnostic categories (p < 0.001), with the exception of specificity for the equivocal category (p = 0.883). Diagnostic accuracy was higher for the consistent category than for the inconsistent and equivocal categories (p < 0.001). Comparisons of pre-training and post-training results revealed that inter-rater agreement was poor and did not improve after training, as assessed by overall agreement (0.450 ± 0.406 vs. 0.461 ± 0.575, p = 0.792), Fleiss's kappa (0.133 ± 0.575 vs. 0.178 ± 0.710, p = 0.405), and intraclass correlation coefficient (ICC; 0.219 vs. 0.276, p = 0.470). CONCLUSIONS: The radiographic diagnostic accuracy and inter-rater agreement were poor when the Berlin radiographic definition was used, and were not significantly improved by the training set of chest radiographs developed by the ARDS Definition Task Force. TRIAL REGISTRATION: The study was registered at ClinicalTrials.gov (registration number NCT01704066 ) on 6 October 2012.
Assuntos
Competência Clínica/normas , Radiografia Torácica/métodos , Síndrome do Desconforto Respiratório/diagnóstico , Ensino/normas , Competência Clínica/estatística & dados numéricos , Feminino , Humanos , Masculino , Variações Dependentes do Observador , Estudos Prospectivos , Radiografia Torácica/estatística & dados numéricos , Reprodutibilidade dos Testes , Síndrome do Desconforto Respiratório/diagnóstico por imagem , Ensino/estatística & dados numéricosRESUMO
OBJECTIVES: This study was designed to test the effectiveness of common carotid artery sonography in comparison with transthoracic echocardiography (TTE) for cardiac output measurements to provide an easier alternative for cardiac output monitoring in the intensive care unit. METHODS: This study included 148 patients who had common carotid artery Doppler examinations and TTE performed within 8 hours of each other, and the cardiac output measurement results were compared with each other. RESULTS: The mean age of the participants ± SD was 56.8 ± 16.2 years, with male patients composing 54.7% of the cohort. There was no significant difference in carotid and TTE cardiac output between different sexes, age groups, patients with and without mechanical ventilation, and primary indication groups. The overall intraclass correlation coefficient between the carotid and TTE cardiac output was 0.537. In patients with septic shock, multiple trauma, and respiratory failure, the intraclass correlation coefficients between TTE and carotid cardiac output were 0.241, 0.061, and 0.095, respectively. CONCLUSIONS: Carotid cardiac output shows moderate agreement with TTE cardiac output; thus, its use may be considered as an alternative for estimating cardiac output in emergencies and when TTE cardiac output is unobtainable. However, in patients with septic shock, multiple trauma, and respiratory failure, the use of carotid cardiac output is not recommended.
Assuntos
Débito Cardíaco/fisiologia , Artéria Carótida Primitiva/diagnóstico por imagem , Artéria Carótida Primitiva/fisiopatologia , Cuidados Críticos/métodos , Ecocardiografia/métodos , Ultrassonografia Doppler/métodos , Idoso , Feminino , Humanos , Unidades de Terapia Intensiva , Masculino , Pessoa de Meia-Idade , Estudos RetrospectivosRESUMO
BACKGROUND: Although the nicotinamide adenine dinucleotide (NAD(+))/CD38/cyclic ADP ribose (cADPR)/Ca(2+) signaling pathway has been shown to regulate intracellular calcium homeostasis and functions in multiple inflammatory processes, its role in sepsis remains unknown. The aim of this study was to determine whether the NAD(+)/CD38/cADPR/Ca(2+) signaling pathway is activated during sepsis and whether an inhibitor of this pathway, 8-Br-cADPR, protects the organs from sepsis-induced damage. MATERIALS AND METHODS: Male Sprague-Dawley rats were subjected to cecal ligation and puncture (CLP) or sham laparotomies. NAD(+), cADPR, CD38, and intracellular Ca(2+) levels were measured in the hearts, livers, and kidneys of septic rats at 0, 6, 12, 24, and 48 h after CLP surgery. Rats were also divided into sham, CLP, and CLP+8-Br-cADPR groups, and the hearts, livers, and kidneys were hematoxylin-eosin-stained and assayed for malondialdehyde and superoxide dismutase activities. RESULTS: NAD(+), cADPR, CD38, and intracellular Ca(2+) levels increased in the hearts, livers, and kidneys of septic rats as early as 6-24 h after CLP surgery. Treatment with 8-Br-cADPR inhibited sepsis-induced intracellular Ca(2+) mobilization, attenuated tissue injury, reduced malondialdehyde levels, and increased superoxide dismutase activity in septic rats. CONCLUSIONS: The NAD(+)/CD38/cADPR/Ca(2+) signaling pathway was activated during sepsis in the CLP rat model. Blocking this pathway with 8-Br-cADPR protected hearts, livers, and kidneys from sepsis-induced damage.
Assuntos
Sinalização do Cálcio/efeitos dos fármacos , ADP-Ribose Cíclica/análogos & derivados , Insuficiência de Múltiplos Órgãos/prevenção & controle , Sepse/complicações , ADP-Ribosil Ciclase/metabolismo , ADP-Ribosil Ciclase 1/metabolismo , Animais , Cálcio/metabolismo , ADP-Ribose Cíclica/metabolismo , ADP-Ribose Cíclica/farmacologia , ADP-Ribose Cíclica/uso terapêutico , Modelos Animais de Doenças , Avaliação Pré-Clínica de Medicamentos , Masculino , Malondialdeído/metabolismo , Glicoproteínas de Membrana/metabolismo , Insuficiência de Múltiplos Órgãos/etiologia , NAD/metabolismo , Distribuição Aleatória , Ratos Sprague-Dawley , Sepse/metabolismo , Superóxido Dismutase/metabolismoRESUMO
OBJECTIVES: To determine the role of neuroglobin in the pathology of sepsis-associated encephalopathy and ascertain if neuroglobin has any protective effects against sepsis-associated encephalopathy. DESIGN: Randomized laboratory animal study. SETTING: Research university animal laboratory. SUBJECTS: Two hundred and forty adult male Sprague-Dawley rats. INTERVENTIONS: Rats received cecal puncture and ligation (or sham) surgery to induce sepsis, then broken up into groups based on whether or not the rat developed sepsis-associated encephalopathy as determined by electroencephalograph and evoked potential recordings. The rats were then left untreated to examine the effect of sepsis-associated encephalopathy on neuroglobin, treated with a neuroglobin antisense nucleotide to block gene expression, or given hemin, a neuroglobin inducer. MEASUREMENTS AND MAIN RESULTS: Following sepsis induction, diagnosis, and treatment, the brains were analyzed for both gross and ultrastructural morphology. Also, neuronal neuroglobin immunoreactivity and apoptosis (via terminal uridine nucleotide end-labeling) were examined. Blood serum levels were then analyzed for neuroglobin, superoxide dismutase, and malondialdehyde levels. We determined that sepsis-associated encephalopathy induces damage evident when examining both gross and ultrastructural morphology, as well as induces neuronal neuroglobin expression. Also, blockade of neuroglobin expression via antisense treatment will exacerbate these pathological effects, while increasing neuroglobin levels via hemin will ameliorate them. Blood analysis found that levels of superoxide dismutase and malondialdehyde mirrored the level of pathology found in the brain, while plasma neuroglobin levels reflected the amount of neuronal neuroglobin immunoreactivity. CONCLUSIONS: We conclude that neuroglobin is involved in the pathogenesis of sepsis-associated encephalopathy and has neuroprotective effects. We also determined that hemin has protective effects against sepsis-associated encephalopathy as well, most probably due to its effect on neuroglobin.
Assuntos
Encefalopatias/etiologia , Globinas/fisiologia , Proteínas do Tecido Nervoso/fisiologia , Sepse/complicações , Animais , Apoptose/fisiologia , Encéfalo/patologia , Encéfalo/ultraestrutura , Encefalopatias/patologia , Encefalopatias/fisiopatologia , Modelos Animais de Doenças , Globinas/biossíntese , Masculino , Malondialdeído/sangue , Microscopia Eletrônica de Transmissão , Proteínas do Tecido Nervoso/biossíntese , Proteínas do Tecido Nervoso/sangue , Neuroglobina , Ratos , Ratos Sprague-Dawley , Sepse/patologia , Sepse/fisiopatologia , Superóxido Dismutase/sangueRESUMO
S100ß and neuron-specific enolase (NSE) are brain injury biomarkers, mainly used in brain trauma, cerebral stroke and hypoxic ischemia encephalopathy. The aim of this study was to study the clinical significance of serum S100ß and NSE in diagnosing sepsis-associated encephalopathy (SAE) and predicting its prognosis. This was a prospective and observational study. Clinical data of septic patients were collected within 24 h after ICU admission from May 2012 to April 2013. We evaluated the level of consciousness twice per day. SAE was defined as cerebral dysfunction in the presence of sepsis that fulfilled the exclusion criteria. The infection biochemical indicators, Glasgow coma scale (GCS) score, acute physiology and chronic health evaluation score II, serum NSE and S100ß were newly measured or evaluated for SAE patients. Finally, hospital mortality, bacteriological categories, length of ICU stay and length of hospital stay were also recorded for all enrolled patients. The data was analyzed with the Chi square test, two-sample t test or Mann-Whitney U test between two groups. The correlation between two factors was analyzed using the Pearson or Spearman analysis. Receiver operating characteristic (ROC) curves were used to determine the ability of S100ß and NSE in diagnosing SAE and predicting the hospital mortality. In addition, cut-off points were obtained from the curves to determine the highest sum of sensitivity and specificity. Of 112 enrolled patients, 48 patients were diagnosed with SAE. The serum S100ß and NSE concentrations in SAE patients were both significantly higher than in non-SAE patients 0.306 (IQR 0.157-0.880) µg/L vs. 0.095 (IQR 0.066-0.177) µg/L, 24.87 (IQR 31.73-12.73) ng/mL vs. 15.49 (IQR 9.88-21.46) ng/mL, P < 0.01]. GCS scores were related more closely to S100ß than NSE (-0.595 vs. -0.337). S100ß levels of 0.131 µg/L diagnosed SAE with 67.2% specificity and 85.4% sensitivity in the ROC curve, the area under the curve was 0.824 (95% confidence interval 0.750-0.898). NSE levels of 24.15 ng/mL diagnosed SAE with 82.8% specificity and 54.2% sensitivity, and the area under the curve was 0.664 (95 % confidence interval 0.561-0.767). In addition, the area under the curve for S100ß for predicting hospital mortality was larger than for NSE (0.730 vs. 0.590). Serum S100ß concentrations in SAE patients were significantly higher than in non-SAE patients. These may be related to the severity of SAE and may predict the outcome of sepsis. The efficacy and sensitivity of serum S100ß in diagnosing SAE were high, but it had a low specificity. Moreover, compared to NSE, serum S100ß was better for both diagnosing SAE and predicting the outcome of sepsis.
Assuntos
Fosfopiruvato Hidratase/sangue , Subunidade beta da Proteína Ligante de Cálcio S100/sangue , Encefalopatia Associada a Sepse/sangue , Encefalopatia Associada a Sepse/diagnóstico , Adulto , Idoso , Biomarcadores/sangue , Feminino , Escala de Coma de Glasgow , Humanos , Masculino , Pessoa de Meia-Idade , PrognósticoRESUMO
AIMS: Sepsis-associated encephalopathy (SAE) often leads to cognitive impairments. However, the pathophysiology of SAE is complex and unclear. Here, we investigated the role of hippocampus (HPC)-prefrontal cortex (PFC) in cognitive dysfunction in sepsis induced by cecal ligation puncture (CLP) in mice. METHODS: The neural projections from the HPC to PFC were first identified via retrograde tracing and viral expression. Chemogenetic activation of the HPC-PFC pathway was shown via immunofluorescent staining of c-Fos-positive neurons in PFC. Morris Water Maze (MWM) and Barnes maze (BM) were used to evaluate cognitive function. Western blotting analysis was used to determine the expression of glutamate receptors and related molecules in PFC and HPC. RESULTS: Chemogenetic activation of the HPC-PFC pathway enhanced cognitive dysfunction in CLP-induced septic mice. Glutamate receptors mediated the effects of HPC-PFC pathway activation in CLP mice. The activation of the HPC-PFC pathway resulted in significantly increased levels of NMDAR, AMPAR, and downstream signaling molecules including CaMKIIa, pCREB, and BDNF in PFC. However, inhibition of glutamate receptors using 2,3-dihydroxy-6-nitro-7-sulphamoyl-benzo (F)quinoxaline (NBQX), which is an α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR inhibitor), or D-2-amino-5-phosphonopentanoate (D-AP5), which is an NMDA receptor antagonist abolished this increase. CONCLUSION: Our study reveals the important role of the HPC-PFC pathway in improving cognitive dysfunction in a mouse model of CLP sepsis and provides a novel pathogenetic mechanism for SAE.
Assuntos
Encefalopatia Associada a Sepse , Sepse , Camundongos , Animais , Aprendizagem Espacial , Sepse/complicações , Sepse/metabolismo , Hipocampo/metabolismo , Córtex Pré-Frontal/metabolismo , Receptores de N-Metil-D-Aspartato/metabolismo , PunçõesRESUMO
Objectives: We aimed to evaluate the association between ß-blocker therapy and mortality in patients with sepsis. Methods: Patients with sepsis were selected from the Medical Information Mart for Intensive Care (MIMIC)-III. Propensity score matching (PSM) was used to balance the baseline differences. A multivariate Cox regression model was used to assess the relationship between ß-blocker therapy and mortality. The primary outcome was the 28-day mortality. Results: A total of 12,360 patients were included in the study, involving 3,895 who received ß-blocker therapy and 8,465 who did not. After PSM, 3,891 pairs of patients were matched. The results showed that ß-blockers were associated with improved 28- (hazards ratio (HR) 0.78) and 90-day (HR 0.84) mortality. Long-acting ß-blockers were associated with improved 28-day survival (757/3627 [20.9%] vs. 583/3627 [16.1%], P < 0.001, HR0.76) and 90-day survival (1065/3627 [29.4%] vs.921/3627 [25.4%], P < 0.001, HR 0.77). Short-acting ß-blocker treatment did not reduce the 28-day and 90-day mortality (61/264 [23.1%] vs. 63/264 [23.9%], P = 0.89 and 83/264 [31.4%] vs. 89/264 [31.7%], P = 0.8, respectively). Conclusions: ß-blockers were associated with improved 28- and 90-day mortality in patients with sepsis and septic shock. Long-acting ß-blocker therapy may have a protective role in patients with sepsis, reducing the 28-day and 90-day mortality. However, short-acting ß-blocker (esmolol) treatment did not reduce the mortality in sepsis.
Assuntos
Sepse , Choque Séptico , Humanos , Pontuação de Propensão , Sepse/tratamento farmacológico , Antagonistas Adrenérgicos beta/uso terapêutico , Choque Séptico/tratamento farmacológico , Estudos RetrospectivosRESUMO
OBJECTIVE: To assess the clinical applicability of focused transthoracic echocardiography (TTE) in intensive care unit (ICU) performed by intensivists and its impacts on clinical managements. METHODS: After 12-hour tutorials and initial cardiac clinical assessments, intensivists performed a focused TTE (2-4 views of 2D, without Doppler or M mode) examination in 88 patients to assess left ventricular function and left ventricular volume status, and rule out local ventricular wall motion abnormalities and significant pericardial effusions. Each investigation was immediately reviewed by an echocardiograph to determine the technical quality of the TTE and the accuracy of the intensivist's interpretation. RESULTS: Intensivists successfully performed a diagnostic focused TTE in 86 patients (97.7%) and interpreted correctly in 75 patients (85.2%). Management including fluid treatment, inotropic agent and vasoactive agent in 22.7% of patients were changed directly based on the focused TTE, 45.5% of patients were provided with valuable information, while 31.8% of them with non-valuable information. The mean focused TTE acquisition time of the intensivist was (11.2±5.2) minutes. CONCLUSIONS: After a brief standard training in using echocardiographic system, intensivists can successfully performed and correctly interpreted a focused TTE for critically ill patients. Our study demonstrates that new information can be provided by focused TTE, which can alter management in a significant number of patients. The present study supports incorporating bedside goal-directed, focused TTE into intensivists' training programs in China.
Assuntos
Ecocardiografia , Unidades de Terapia Intensiva , Sistemas Automatizados de Assistência Junto ao Leito , Adulto , Idoso , Idoso de 80 Anos ou mais , Estado Terminal , Ecocardiografia Doppler , Estudos de Viabilidade , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Médicos , Estudos ProspectivosRESUMO
OBJECTIVE: To observe the effects of adenovirus borne IΚB gene, an inhibitor of nuclear factor-ΚB (NF-ΚB), infused via central vein, to treat infectious acute lung injury (ALI) in rats. METHODS: According to random number table method, 30 pathogen-free Sprague-Dawley (SD) rats were randomly divided into three groups: sham group, ALI model group, IΚB gene treatment group, with 10 rats in each group. The rats of IΚB gene treatment group were infused 1 ml adenovirus borne IΚB gene (titre: 1×10(9)pfu ), the rats of sham group and ALI model group were infused 1 ml normal saline through central vein. Subsequently, the rats of ALI model group and the IΚB gene treatment group were given 1 ml lipopolysaccharide (LPS, 5 ml/kg) through tail vein to reproduce model of ALI. On the other hand, the rats of sham group were given 1 ml normal saline through tail vein. Blood gas analysis, the ratio of wet to dry weight (W/D) of lung, plasma contents of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6), and protein expression of NF-ΚBp65 in lung tissue were determined, the pathobiological changes in lung tissue were microscopically observed and the scores of lung injury were calculated after 7 days. RESULTS: The rats in three groups survived, except 1 rat died in ALI model group.Blood pH and partial pressure of arterial carbon dioxide (PaCO(2)) showed no obviously differences among three groups. Partial pressure of arterial oxygen (PaO(2) ) was highest in sham group and the lowest in ALI model group. The plasma content of TNF-α (µg/L) and IL-6 (ng/L ) in ALI model group were obviously higher than those in sham group (TNF-α: 5.20±1.09 vs. 3.01±0.46; IL-6: 540.28±100.78 vs. 214.45±61.37, both P<0.05). The plasma content of TNF-α and IL-6 in IΚB gene treatment group were obviously lower than those in ALI model group (TNF-α: 3.70±0.96 vs. 5.20±1.09, IL-6: 356.49±60.58 vs. 540.28±100.78, both P<0.05), and TNF-α content had restored to the level observed in sham group. The ratio of W/D of lung was lowest in sham group (4.49±0.36) and highest in ALI model group (5.78±0.43), and that of IΚB gene treatment group (5.33±0.38) was lower than that of ALI group. The score of lung injury was lowest in sham group (0.17±0.41) and highest in ALI model group (2.29±0.76), and that of IΚB gene treatment group (1.57±0.53) was lower than that of ALI group. The scale of NF-ΚBp65 immunohistochemistry was lowest in sham group (1.00±0.89) and highest in ALI model group (9.43±1.13), and that of IΚB gene treatment group (4.00±1.15) was lower than the latter. The differences of all the above parameters in three groups were statistically significant (all P<0.05 ). CONCLUSION: Increased expression of IΚB gene by an infusion of adenovirus borne IΚB gene through central vein can lower the levels of pro-inflammatory factors, such as TNF-α and IL-6, restrain the NF-ΚB activation, reduce lung water, alleviate alveolar collapse and lung consolidation in ALI in rats, thus lung injury is ameliorated.
Assuntos
Lesão Pulmonar Aguda/terapia , Terapia Genética/métodos , Proteínas I-kappa B/genética , Lesão Pulmonar Aguda/metabolismo , Adenoviridae/genética , Animais , Modelos Animais de Doenças , Proteínas I-kappa B/administração & dosagem , Interleucina-6/metabolismo , Masculino , Inibidor de NF-kappaB alfa , NF-kappa B/metabolismo , Ratos , Ratos Sprague-Dawley , Fator de Necrose Tumoral alfa/metabolismoRESUMO
We investigated the role of dynamic changes of serum levels S100B protein in brain injury and poor outcome of sepsis. This is a prospective cohort study designed to include 104 adult patients with sepsis who are admitted to ICU from Jan 2015 to Aug 2016. Sepsis was defined as sepsis 3.0. Patients with a GCS score of <15, or at least one positive CAM-ICU score were thought to have brain dysfunction. 59 patients were diagnosed with SAE and the rest 45 patients were diagnosed with non-SAE. Serum S100B was measured on day 1 and 3 after ICU admission. Primary outcomes included brain dysfunction and 28-day/180-day mortality. The SAE group showed a significantly higher APACHE II score, SOFA scores, length of ICU stay, 28-day and 180-day mortality, serum S100B levels on day 1 and day 3. S100B levels on day 1 of 0.226 µg/L were diagnostic for SAE with 80.0% specificity and 66.1% sensitivity, and the area under (AUC) the curve was 0.728, S100B levels on day 3 of 0.144 µg/L were diagnostic for SAE with 84.44% specificity and 69.49% sensitivity, and the AUC was 0.819. In addition, the AUC for S100B on day 3 for predicting 180-day mortality was larger than for S100B on day 1 (0.731 vs. 0.611). Multiple logistic regression analysis showed that S100B3 (p = 0.001) but not S100B1 (p = 0.927) were independently correlated with SAE. Kaplan-Meier survival analysis showed that patients with S100B levels higher than 0.144 µg/L had a lower probability of survival at day 180. There were more patients with encephalopathy and a higher 28-day or 180-day mortality in the ΔS100B + group than in the ΔS100B- group. Multiple logistic regression analysis showed that SAE and IL-6 on day 3 were independently correlated with S100B dynamic increase. These findings suggest that elevated serum S100B levels on day 3 and the dynamic changes of serum S100B levels from day three to one were more associated with brain dysfunction and mortality than that on day 1 in patients with sepsis.
Assuntos
Lesões Encefálicas/sangue , Interleucina-6/sangue , Subunidade beta da Proteína Ligante de Cálcio S100/sangue , Encefalopatia Associada a Sepse/sangue , Lesões Encefálicas/epidemiologia , Lesões Encefálicas/patologia , Feminino , Humanos , Estimativa de Kaplan-Meier , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Prognóstico , Curva ROC , Encefalopatia Associada a Sepse/epidemiologia , Encefalopatia Associada a Sepse/patologiaRESUMO
Sepsis-induced inflammatory damage is a crucial cause of acute kidney injury (AKI), and AKI is an ecumenical fearful complication in approximately half of patients with sepsis. CCAAT/enhancer-binding protein ß (C/EBPß) plays roles in regulating acute phase responses and inflammation. However, the role and mechanism of C/EBPß in AKI are unclear. LPS combined with ATP-treated renal epithelial cells HK2 and cecal ligation-peferation (CLP)-mice were used as models of AKI in vitro and in vivo. Cell damage, the secretion of interleukin-1 beta (IL-1ß), IL-18 and cysteinyl aspartate specific proteinase 1 (caspase-1) activity were tested by LDH, ELISA assay and flow cytometry analysis, respectively. The expression levels of TFAM, C/EBPß, and pyroptosis-related molecules were tested by qRT-PCR and Western blotting. Chromatin immunoprecipitation (ChIP) assessed the interaction between C/EBPß with TFAM. Hematoxylin-Eosin (H&E) staining detected pathological changes of kidney tissues, and immunohistochemistry measured TFAM and C/EBPß in mice kidney tissues. C/EBPß or TFAM were up-regulated in LPS combined with ATP -induced HK2 cells. Knockdown of C/EBPß could suppress cell injury and the secretion of IL-1ß and IL-18 induced by LPS combined with ATP. Furthermore, C/EBPß up-regulated the expression levels of TFAM via directly binding to TFAM promoter. Overexpression of TFAM reversed the effects of C/EBPß deficiency on pyroptosis. Knockdown of C/EBPß could inhibit NLRP3 inflammasome-mediated caspase-1 signaling pathway by inactivating TFAM/RAGE pathway. It was further confirmed in the AKI mice that C/EBPß and TFAM promoted AKI by activating NLRP3-mediated pyroptosis. The interaction of between C/EBPß and TFAM facilitated pyroptosis by activating NLRP3/caspase-1 signal axis, thereby promoting the occurrence of AKI.
Assuntos
Injúria Renal Aguda/metabolismo , Injúria Renal Aguda/patologia , Proteína beta Intensificadora de Ligação a CCAAT/metabolismo , Proteínas de Ligação a DNA/metabolismo , Proteínas de Grupo de Alta Mobilidade/metabolismo , Inflamassomos/metabolismo , Proteínas Mitocondriais/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Piroptose , Fatores de Transcrição/metabolismo , Trifosfato de Adenosina , Animais , Caspase 1/metabolismo , Linhagem Celular , Humanos , Inflamação/patologia , Lipopolissacarídeos , Masculino , Camundongos Endogâmicos C57BL , Regiões Promotoras Genéticas/genética , Ligação Proteica , Receptor para Produtos Finais de Glicação Avançada/metabolismo , Transdução de SinaisRESUMO
OBJECTIVE: To study the effect of low molecular weight heparin (LMWH) in the treatment of severe acute pancreatitis (SAP). METHODS: A total of 265 SAP patients were randomly divided into two groups: firstly, the conventional treatment group (C group, n = 130; and secondly the conventional treatment plus the LMWH treatment group (LT group, n = 135). The clinical parameters, laboratory parameters and computed tomography (CT) score of pancreatic necrosis (CTSPN) in the two groups were compared. RESULTS: On admission, all the clinical parameters, laboratory parameters and CTSPN in the two groups were not significantly different (p > 0.05). However, after treatment, in LT group, the clinical presentation improvement rate and laboratory parameters improvement were significantly higher than those in C group (p < 0.05-0.01), and the acute physiology and chronic health evaluation (APACHE) II score, complication rate, mortality and mean hospital stay in LT group were obviously lower than those in C group (p < 0.05-0.01). The CT score in LT group was much lower than that in C group (p < 0.05). Two weeks after treatment FBI decreased obviously in C group, but not in LT group, and no haemorrhagic complications occurred. CONCLUSIONS: LMWH can enhance the effect of conventional treatment for SAP, and can markedly decrease the mortality of SAP. LMWH is a simple, safe, economic and effective method for treatment of SAP. It is can be used in every hospital.
Assuntos
Anticoagulantes/uso terapêutico , Heparina de Baixo Peso Molecular/uso terapêutico , Pancreatite/tratamento farmacológico , APACHE , Doença Aguda , Adolescente , Adulto , Idoso , Criança , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Insuficiência de Múltiplos Órgãos/epidemiologia , Estudos Prospectivos , Adulto JovemRESUMO
OBJECTIVE: To investigate the effect of apoptosis of CD4+ CD25+ regulatory T cells (Tregs) on proliferation as well as secretory function of effector T cells (Teff) and potential influence of Xuebijing injection on them in septic rats. METHODS: A sepsis model was reproduced by cecal ligation puncture (CLP), and Wistar rats were randomly divided into the control group (n = 8), sham-operated group (n = 8), CLP group (n = 8), and Xuebijing injection treatment group (n = 8). CD4+ CD25+ Tregs in each group were separated by immunomagnetic beads isolate system on day 3, the apoptosis rate, expression of forkhead/winged helix transcription factor p3 (Foxp3) and cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) on Tregs were analyzed by flow cytometry, and secretion levels of interleukin (IL)-10 from Tregs were measured by ELISA. Following co-culture of CD4+ CD25+ Treg with CD4+ CD25- T cells (1:1) for 68 hours, proliferative activity of Teff was determined by MTT, and IL-2/sIL-2R alpha levels were measured by ELISA. RESULTS: The apoptosis rate of Tregs in control group was 12.03% +/- 0.89%, which was not significantly different from sham-operated group 9.48% +/- 2.17%. The apoptosis rate of Tregs in CLP group 5.87% +/- 0.44% was lower than that in control group (P < 0.01), and treatment with Xuebijing injection markedly enhanced the apoptosis of Tregs 27.29% +/- 2.48%. Compared to CLP group, expression of Foxp3, CTLA-4, and the secretion of IL-10 of Treg were significantly lowered in Xuebijing injection group (all P < 0.01). The Teff proliferative activity in response to ConA, and IL-2 levels of Teff in CLP group were significantly suppressed compared with control group (P < 0.01), and secretion of sIL-2R alpha in the supernatants was much higher than that of the control group. In comparison to the CLP group, inhibitory rate of Teff proliferative activity and the sIL-2R alpha levels were significantly decreased, while the secretion of IL-2 was increased in Xuebijing injection group (P <0.01). CONCLUSION: CD4+ CD25+ Tregs could markedly upregulate the suppressive function on Teff in sepsis, and treatment with Xuebijing injection effectively enhanced apoptosis of Tregs, thereby down-regulating the suppression on Teff.
Assuntos
Apoptose , Proliferação de Células , Medicamentos de Ervas Chinesas/uso terapêutico , Sepse/imunologia , Linfócitos T Reguladores/imunologia , Animais , Modelos Animais de Doenças , Masculino , Ratos , Ratos Wistar , Sepse/tratamento farmacológico , Linfócitos T/imunologia , Linfócitos T/patologia , Linfócitos T Reguladores/patologiaRESUMO
OBJECTIVE: To evaluate the influence of apoptosis of CD4(+)CD25(+) regulatory T lymphocyte (Treg) on polarization of helper T lymphocyte (Th) and effect of Xuebijing injection in septic rats. METHODS: A sepsis model was reproduced by cecal ligation and puncture (CLP). Wistar rats were randomly divided into the normal group (n=8), sham operation group (n=8), model group (n=8) and Xuebijing injection treatment group (n=8). CD4(+)CD25(+)Tregs in each group were separated by immunomagnetic beads isolation system on day 3, the apoptotic rates, expression of forkhead/winged helix transcription factor p3 (Foxp3) as well as cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) of Treg were analyzed by flow cytometry, and the secretion level of interleukin-10 (IL-10) of Tregs was determined by enzyme linked immunosorbent assay (ELISA) after culturing Tregs for 12 hours. Interferon-gamma (IFN-gamma), IL-4, and IL-17 levels, which were respectively secreted by Th1, Th2 and Th17, were measured by ELISA in the supernatant after CD4(+)CD25(+)Tregs were co-cultured with CD4(+)CD25(-)T lymphocytes for 68 hours. RESULTS: The apoptosis rate of the normal group was (12.03+/-0.89)%, which was not significantly different compared with the sham operation group [(9.48+/-2.17)%]. The apoptosis rate of model group [(5.87+/-0.44)%] was much lower than that of the normal and sham operation groups, while the Xuebijing injection treatment group [(27.29+/-2.48)%] had the highest apoptosis rate compared to others (all P<0.01) . The expression of Foxp3, CTLA-4, and the secretion of IL-10 of Treg were negatively correlated with the apoptosis rates, correlation coefficients (r) respectively were -0.878 (P=0.042), -0.877 (P=0.042), and -0.743 (P=0.010). The secretion of IFN-gamma, IL-4 and ratio of IFN-gamma/IL-4 in model group were significantly elevated compared with the normal group [IFN-gamma: (254.70+/-44.88) ng/L vs. (0.68+/-0.78) ng/L , IL-4: (8.82+/-0.61) ng/L vs. (3.48+/-0.98) ng/L, ratio of IFN-gamma/IL-4: 30.28+/- 4.87 vs. 0.23+/-0.30, all P<0.01], and secretion of IFN-gamma as well as ratio of IFN-gamma/IL-4 were markedly higher in Xuebijing injection treatment group [(491.54+/-84.28) ng/L, 45.31+/-8.01, respectively] than in model group (P<0.01 and P<0.05). No marked change in IL-17 levels was noted in various groups (all P>0.05) . CONCLUSION: Due to the apoptosis of Treg, the suppression function of CD4(+)CD25(+)Tregs on CD4(+)T lymphocyte appears to be abated, and treatment with Xuebijing injection could effectively enhance the apoptosis of Tregs, mediating the response of shifting Th2 to Th1.
Assuntos
Sepse/imunologia , Linfócitos T Auxiliares-Indutores/imunologia , Linfócitos T Reguladores/imunologia , Animais , Antígenos CD/metabolismo , Apoptose/efeitos dos fármacos , Antígeno CTLA-4 , Células Cultivadas , Técnicas de Cocultura , Modelos Animais de Doenças , Medicamentos de Ervas Chinesas/farmacologia , Fatores de Transcrição Forkhead/metabolismo , Interferon gama/metabolismo , Interleucinas/metabolismo , Masculino , Distribuição Aleatória , Ratos , Ratos Wistar , Sepse/tratamento farmacológico , Sepse/metabolismo , Sepse/patologia , Linfócitos T Reguladores/metabolismo , Linfócitos T Reguladores/patologiaRESUMO
PURPOSE: We investigated the role of serum Glial Fibrillary Acidic Protein (GFAP) and Ubiquitin C-Terminal Hydrolase-L1 (UCH-L1) in diagnosis of sepsis-associated encephalopathy(SAE), predicting prognosis and long-term quality of life with patients of sepsis. MATERIALS AND METHODS: This is a prospective single center study entailed 105 patients whosuffered from sepsis from Jan 2015 to Aug 2016. Serum concentrations of GFAP and UCH-L1 for diagnosis of SAE and predicting prognosis and long-term quality of life with patients of sepsis were analyzed. RESULTS: The serum concentrations of GFAP and UCH-L1 were higher in SAE group than in no-SAE group (pâ¯<â¯.001). GFAP and UCH-L1 produced an AUC of 0.824 and 0.812 respectively for diagnosis of SAE with optimal cut-off values 0.532â¯ng/ml and 7.72â¯ng/ml respectively. The optimal cut-off values of GFAP and UCH-L1 to distinguish patients with survivors from non-survivors were 0.536â¯ng/ml and 8.06â¯ng/ml with an area under the curve of 0.773 and 0.746. Patients with a higher GFAP levels had worse long-term usual activities and patients with a higher UCH-L1 levels had more long-term pain (Pâ¯=â¯.026). CONCLUSIONS: Serum concentrations GFAP and UCH-L1 early elevated and associated with sepsis-associated encephalopathy, poor prognosis and quality of life.
Assuntos
Proteína Glial Fibrilar Ácida/metabolismo , Encefalopatia Associada a Sepse/diagnóstico , Ubiquitina Tiolesterase/metabolismo , Biomarcadores/metabolismo , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico , Estudos Prospectivos , Qualidade de Vida , Encefalopatia Associada a Sepse/sangueRESUMO
BACKGROUND: Studies have reported mitophagy activation in renal tubular epithelial cells (RTECs) in acute kidney injury (AKI). Phosphatase and tensin homolog-induced putative kinase 1 (PINK1) and E3 ubiquitin-protein ligase Parkin are involved in mitophagy regulation; however, little is known about the role of PINK1-Parkin mitophagy in septic AKI. Here we investigated whether the PINK1-Parkin mitophagy pathway is involved in septic AKI and its effects on cell apoptosis in vitro and on renal functions in vivo. METHODS: Mitophagy-related gene expression was determined using Western blot assay in human RTEC cell line HK-2 stimulated with bacterial lipopolysaccharide (LPS) and in RTECs from septic AKI rats induced by cecal ligation and perforation (CLP). Autophagy-related ultrastructural features in rat RTECs were observed using electron microscopy. Gain- and loss-of-function approaches were performed to investigate the role of the PINK1-Parkin pathway in HK-2 cell mitophagy. Autophagy activators and inhibitors were used to assess the effects of mitophagy modulation on cell apoptosis in vitro and on renal functions in vivo. RESULTS: LPS stimulation could significantly induce LC3-II and BECN-1 protein expression (LC3-II: 1.72â±â0.05 vs. 1.00â±â0.05, Pâ<â0.05; BECN-1: 5.33â±â0.57 vs. 1.00â±â0.14, Pâ<â0.05) at 4 h in vitro. Similarly, LC3-II, and BECN-1 protein levels were significantly increased and peaked at 2 h after CLP (LC3-II: 3.33â±â0.12 vs. 1.03â±â0.15, Pâ<â0.05; BECN-1: 1.57â±â0.26 vs. 1.02â±â0.11, Pâ<â0.05) in vivo compared with those after sham operation. Mitochondrial deformation and mitolysosome-mediated mitochondria clearance were observed in RTECs from septic rats. PINK1 knockdown significantly attenuated LC3-II protein expression (1.35â±â0.21 vs. 2.38â±â0.22, Pâ<â0.05), whereas PINK1 overexpression markedly enhanced LC3-II protein expression (2.07â±â0.21 vs. 1.29â±â0.19, Pâ<â0.05) compared with LPS-stimulated HK-2 cells. LPS-induced proapoptotic protein expression remained unchanged in autophagy activator-treated HK-2 cells and was significantly attenuated in PINK1-overexpressing cells, but was remarkably upregulated in autophagy inhibitor-treated and in PINK1-depleted cells. Consistent results were observed in flow cytometric apoptosis assay and in renal function indicators in rats. CONCLUSION: PINK1-Parkin-mediated mitophagy might play a protective role in septic AKI, serving as a potential therapeutic target for septic AKI.
Assuntos
Injúria Renal Aguda/fisiopatologia , Mitofagia/fisiologia , Proteínas Quinases/fisiologia , Ubiquitina-Proteína Ligases/fisiologia , Animais , Proteína Beclina-1/análise , Células Cultivadas , Células Epiteliais/fisiologia , Humanos , Túbulos Renais/citologia , Lipopolissacarídeos , Proteínas Associadas aos Microtúbulos/análise , Ratos , Ratos Sprague-Dawley , Sepse/fisiopatologiaRESUMO
CD4+ T cells have been divided into different subsets, essentially based on the cytokines they produce. These subsets include T helper cell (Th)1, Th2 and regulatory T cells (Treg). Recently, another subpopulation of T cells has been identified, and it was named as Th17, which is characterized by the release of interleukin (IL)-17. The discovery of Th17 cells is not only a challenge to some traditional theories, such as the differentiation of CD4+ T cells and the drift theory of Th1/Th2, but also provides new research approach to inflammatory and immunologic diseases, especially, the balance among Th1, Th2, Treg and Th17 cells may play an important role in maintaining the normal immune response.
Assuntos
Células Th17 , Animais , Humanos , Interleucina-17/metabolismo , Células Th17/metabolismoRESUMO
BACKGROUND: The CD38/cADPR pathway has been found to play roles in various inflammatory conditions. However, whether CD38 plays a protective or detrimental effect in the central nervous system (CNS) is controversial. The aim of this study was to determine the effect of CD38/cADPR pathway in sepsis associated brain injury. MATERIALS AND METHODS: Male Sprague-Dawley rats were undergone cecal ligation and puncture (CLP) or sham laparotomies. NAD+, cADPR and CD38 were measured in the hippocampus of septic rats at 0, 6, 12, 24, and 48h after CLP surgery. Rats were divided into the sham, CLP group, CLP+ CD38 expression lentivirus (CLP+ CD38 LV), CLP+ CD38 interference lentivirus (CLP+ CD38 Ri), CLP+ negative control lentivirus (CLP+NC) and the CLP+8-Br-cADPR groups. The Western blots of Bcl-2, Bax and iNOS, TUNEL assays, malondialdehyde (MDA) and superoxide dismutase (SOD) assays, transmission electron microscope analysis were performed in the hippocampus of rats. RESULTS: NAD+, cADPR and CD38 levels increased significantly in the hippocampus of septic rats as early as 12-24h after CLP surgery. CD38 knockdown or blocking cADPR with 8-Br-cADPR significantly reduced apoptosis, MDA and SOD activity, iNOS expression and ultrastructural morphology damages in the hippocampus of septic rats. CONCLUSIONS: In this study, we found that the CD38/cADPR pathway was activated in sepsis associated brain injury. Blocking this pathway protected the hippocampus from apoptosis, oxidative stress and ultrastructural morphology damages in septic rats.