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As one of the post-transcriptional regulatory mechanisms, uncoupling of transcription and translation plays an essential role in development and adulthood physiology. However, it remains elusive how thousands of mRNAs get translationally silenced while stability is maintained for hours or even days before translation. In addition to oocytes and neurons, developing spermatids display significant uncoupling of transcription and translation for delayed translation. Therefore, spermiogenesis represents an excellent in vivo model for investigating the mechanism underlying uncoupled transcription and translation. Through full-length poly(A) deep sequencing, we discovered dynamic changes in poly(A) length through deadenylation and re-polyadenylation. Deadenylation appeared to be mediated by microRNAs (miRNAs), and transcripts with shorter poly(A) tails tend to be sequestered into ribonucleoprotein (RNP) granules for translational repression and stabilization. In contrast, re-polyadenylation might allow for translocation of the translationally repressed transcripts from RNP granules to polysomes. Overall, our data suggest that miRNA-dependent poly(A) length control represents a previously unreported mechanism underlying uncoupled translation and transcription in haploid male mouse germ cells.
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MicroRNAs , Poli A , Animais , Haploidia , Masculino , Camundongos , MicroRNAs/genética , MicroRNAs/metabolismo , Poli A/metabolismo , Biossíntese de Proteínas , RNA Mensageiro/metabolismo , Espermátides/metabolismoRESUMO
INTRODUCTION: Nowadays,the mechanical ventilation (MV) aims to rest the respiratory muscles while providing adequate gas exchange, and it has been a part of basic life support during general anesthesia as well as in critically ill patients with and without respiratory failure. However, MV itself has the potential to cause or worsen lung injury, which is also known as ventilator-induced lung injury (VILI). Thus, the early diagnosis of VILI is of great importance for the prevention and treatment of VILI. OBJECTIVE: This study aimed to investigate the metabolomes in the lung and plasma of mice receiving mechanical ventilation (MV). METHODS: Healthy mice were randomly assigned into control group; (2) high volume tidal (HV) group (30 ml/kg); (3) low volume tidal (LV) group (6 ml/kg). After ventilation for 4 h, mice were sacrificed and the lung tissue and plasma were collected. The lung and plasma were processed for the metabolomics analysis. We also performed histopathological examination on the lung tissue. RESULTS: We detected moderate inflammatory damage with alveolar septal thickening in the HV group compared with the normal and LV groups.The metabolomics analysis results showed MV altered the metabolism which was characterized by the dysregulation of γ-amino butyric acid (GABA) system and urea cycle (desregulations in plasma and lung guanidinosuccinic acid, argininosuccinic acid, succinic acid semialdehyde and lung GABA ), Disturbance of citric acid cycle (CAC) (increased plasma glutamine and lung phosphoenol pyruvate) and redox imbalance (desregulations in plasma and/or lung ascorbic acid, chenodeoxycholic acid, uric acid, oleic acid, stearidonic acid, palmitoleic acid and docosahexaenoic acid). Moreover, the lung and plasma metabolomes were also significantly different between LV and HV groups. CONCLUSIONS: Some lung and plasma metabolites related to the GABA system and urea cycle, citric acid cycle and redox balance were significantly altered, and they may be employed for the evaluation of VILI and serve as targets in the treatment of VILI.
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Metabolômica , Lesão Pulmonar Induzida por Ventilação Mecânica , Animais , Pulmão/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Ureia/metabolismo , Lesão Pulmonar Induzida por Ventilação Mecânica/metabolismo , Lesão Pulmonar Induzida por Ventilação Mecânica/patologia , Lesão Pulmonar Induzida por Ventilação Mecânica/prevenção & controle , Ácido gama-Aminobutírico/metabolismoRESUMO
Sepsis is a global disease burden, and approximately 40% of cases develop acute lung injury (ALI). Bone marrow mesenchymal stromal cells (BMSCs) and their exosomes are widely used in treating a variety of diseases including sepsis. As an acute phase protein, serum amyloid A1 (SAA1) regulates inflammation and immunity. However, the role of SAA1 in BMSCs-exosomes in septic lung injury remains to be elucidated. Exosomes derived from serum and BMSCs were isolated by ultracentrifugation. SAA1 was silenced or overexpressed in mouse BMSCs using lentiviral plasmids, containing either SAA1-targeting short interfering RNAs or SAA1 cDNA. Sepsis was induced by cecal ligation and puncture (CLP). LPS was used to induce ALI in mice. Mouse alveolar macrophages were isolated by flow cytometry. Levels of SAA1, endotoxin, TNF-α, and IL-6 were measured using commercial kits. LPS internalization was monitored by immunostaining. RT-qPCR or immunoblots were performed to test gene and protein expressions. Serum exosomes of patients with sepsis-induced lung injury had significantly higher levels of SAA1, endotoxin, TNF-α, and IL-6. Overexpression of SAA1 in BMSCs inhibited CLP- or LPS-induced lung injury and decreased CLP- or LPS-induced endotoxin, TNF-α, and IL-6 levels. Administration of the SAA1 blocking peptide was found to partially inhibit SAA1-induced LPS internalization by mouse alveolar macrophages and reverse the protective effect of SAA1. In conclusion, BMSCs inhibit sepsis-induced lung injury through exosomal SAA1. These results highlight the importance of BMSCs, exosomes, and SAA1, which may provide novel directions for the treatment of septic lung injury.
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Lesão Pulmonar Aguda , Células-Tronco Mesenquimais , Sepse , Proteína Amiloide A Sérica , Animais , Camundongos , Lesão Pulmonar Aguda/induzido quimicamente , Células da Medula Óssea/metabolismo , Interleucina-6/metabolismo , Lipopolissacarídeos/farmacologia , Fator de Necrose Tumoral alfa/metabolismo , Proteína Amiloide A Sérica/genética , ExossomosRESUMO
CONTEXT: Salidroside, a compound extracted from Rhodiola rosea L. (Crassulaceae), possesses many beneficial pathological effects. OBJECTIVE: To explore the effect of salidroside on ventilator-induced lung endothelial dysfunction in vivo and in vitro. MATERIALS AND METHODS: In vivo, male ICR mice were divided into sham, ventilation, salidroside, and ventilation plus salidroside groups. The mice were ventilated for 4 h, salidroside (50 mg/kg) was administrated intraperitoneally before ventilation, dexamethasone (Dex) (5 mg/kg) was used as a positive control. In vitro, mouse lung vascular endothelial cells (MLVECs) were treated with salidroside, MMP-9 siRNA, and BAY11-7082 (10 µM), and then exposed to cyclic stretch for 4 h. Afterward, lung tissues and MLVECs were collected for further analysis. RESULTS: Salidroside pre-treatment significantly reversed the expression of vascular endothelial cadherin (VE-cadherin) and zonula occluden-1 (ZO-1) proteins in cyclic stretch-treated MLVECs (0.46 ± 0.09 vs. 0.80 ± 0.14, 0.49 ± 0.05 vs. 0.88 ± 0.08) and ventilated lung tissues (0.56 ± 0.06 vs. 0.83 ± 0.46, 0.49 ± 0.08 vs. 0.80 ± 0.12). The results further indicated that salidroside inhibited the expression of matrix metalloproteinase-9 (MMP-9), whereas knockdown of its expression restored the expression levels of VE-cadherin (0.37 ± 0.08 vs. 0.85 ± 0.74) and ZO-1 (0.48 ± 0.08 vs. 0.81 ± 0.11) in stretched MLVECs. Meanwhile, salidroside inhibited the NF-κB signalling pathway and alleviated lung injury. CONCLUSIONS: Salidroside protected against stretch-induced endothelial barrier function, improving lung injury after ventilation. Thus, salidroside may be a promising therapeutic agent for patients with MV-induced lung injury.
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Células Endoteliais/efeitos dos fármacos , Glucosídeos/farmacologia , Metaloproteinase 9 da Matriz/genética , Fenóis/farmacologia , Lesão Pulmonar Induzida por Ventilação Mecânica/prevenção & controle , Animais , Dexametasona/farmacologia , Células Endoteliais/patologia , Técnicas de Silenciamento de Genes , Glucosídeos/isolamento & purificação , Masculino , Camundongos , Camundongos Endogâmicos ICR , NF-kappa B/metabolismo , Fenóis/isolamento & purificação , Rhodiola/química , Transdução de Sinais/efeitos dos fármacosRESUMO
OBJECTIVES: Mechanical ventilation can induce lung fibrosis. This study aimed to investigate whether ventilator-induced lung fibrosis was associated with endothelial-mesenchymal transition and to uncover the underlying mechanisms. DESIGN: Randomized, controlled animal study and cell culture study. SETTING: University research laboratory. SUBJECTS: Adult male Institute of Cancer Research, NACHT, LRR, and PYD domains-containing protein 3 (NLRP3) knockout and wild-type mice. Primary cultured mouse lung vascular endothelial cells. INTERVENTIONS: Institute of Cancer Research, NLRP3 knockout and wild-type mice were subjected to mechanical ventilation (20 mL/kg) for 2 hours. Mouse lung vascular endothelial cells were subjected to cyclic stretch for 24 hours. MEASUREMENTS AND MAIN RESULTS: Mice subjected to mechanical ventilation exhibited increases in collagen deposition, hydroxyproline and type I collagen contents, and transforming growth factor-ß1 in lung tissues. Ventilation-induced lung fibrosis was associated with increased expression of mesenchymal markers (α smooth muscle actin and vimentin), as well as decreased expression of endothelial markers (vascular endothelial-cadherin and CD31). Double immunofluorescence staining showed the colocalization of CD31/α smooth muscle actin, CD31/vimentin, and CD31/fibroblast-specific protein-1 in lung tissues, indicating endothelial-mesenchymal transition formation. Mechanical ventilation also induced NLRP3 inflammasome activation in lung tissues. In vitro direct mechanical stretch of primary mouse lung vascular endothelial cells resulted in similar NLRP3 activation and endothelial-mesenchymal transition formation, which were prevented by NLRP3 knockdown. Furthermore, mechanical stretch-induced endothelial-mesenchymal transition and pulmonary fibrosis were ameliorated in NLRP3-deficient mice as compared to wild-type littermates. CONCLUSIONS: Mechanical stretch may promote endothelial-mesenchymal transition and pulmonary fibrosis through a NLRP3-dependent pathway. The inhibition of endothelial-mesenchymal transition by NLRP3 inactivation may be a viable therapeutic strategy against pulmonary fibrosis associated with mechanical ventilation.
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Modelos Animais de Doenças , Endotélio Vascular/fisiopatologia , Inflamassomos/fisiologia , Pulmão/irrigação sanguínea , Mecanotransdução Celular/fisiologia , Mesoderma/fisiopatologia , Proteína 3 que Contém Domínio de Pirina da Família NLR/fisiologia , Fibrose Pulmonar/fisiopatologia , Animais , Células Cultivadas , Células Endoteliais/fisiologia , Camundongos , Camundongos Endogâmicos ICR , Camundongos KnockoutRESUMO
BACKGROUND: Salidroside (SDS) is the main effective ingredient of Rhodiola rosea L with a variety of pharmacologic properties. We aim to investigate the effects of SDS on ventilation induced lung injury (VILI) and explore the possible underlying molecular mechanism. METHODS: Lung injury was induced in male ICR mice via mechanical ventilation (30 ml/kg) for 4h. The mice were divided in four groups:(1) Control group; (2) Ventilation group; (3) SDS group; (4) Ventilation with SDS group. SDS (50 mg/kg) was injected intraperitoneally 1h before operation. Mouse lung vascular endothelial cells (MLVECs) were subjected to cyclic stretch for 4h. RESULTS: It was found that SDS attenuated VILI as shown in HE staining, cell count and protein content levels in BAL fluid, W/D and Evans blue dye leakage into the lung tissue. SDS treatment inhibited the activation of NLRP3 inflammasome and subsequent caspase-1 cleavage as well as interleukin (IL)-1ß secretion both in vivo and in vitro. Moreover, SDS administration up-regulated SIRT1 expression. Importantly, knockdown of SIRT1 reversed the inhibitory effect of SDS on NLRP3 inflammasome activation. CONCLUSIONS: Taken together, these findings indicate that SDS may confer protection against ventilation induced lung injury via SIRT1-de-pendent inhibition of NLRP3 inflammasome activation.
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Glucosídeos/farmacologia , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Fenóis/farmacologia , Sirtuína 1/metabolismo , Regulação para Cima/efeitos dos fármacos , Animais , Líquido da Lavagem Broncoalveolar/química , Caspase 1/metabolismo , Células Cultivadas , Células Endoteliais/citologia , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/metabolismo , Ensaio de Imunoadsorção Enzimática , Glucosídeos/uso terapêutico , Inflamassomos/metabolismo , Interleucina-1beta/análise , Interleucina-1beta/metabolismo , Pulmão/metabolismo , Pulmão/patologia , Masculino , Camundongos , Camundongos Endogâmicos ICR , Proteína 3 que Contém Domínio de Pirina da Família NLR/antagonistas & inibidores , Fenóis/uso terapêutico , Interferência de RNA , RNA Interferente Pequeno/metabolismo , Sirtuína 1/antagonistas & inibidores , Sirtuína 1/genética , Estresse Mecânico , Lesão Pulmonar Induzida por Ventilação Mecânica/metabolismo , Lesão Pulmonar Induzida por Ventilação Mecânica/patologia , Lesão Pulmonar Induzida por Ventilação Mecânica/prevenção & controleRESUMO
Chronic exposure to the hypobaric hypoxia environment of plateau could influence human cognitive behaviours which are supported by dynamic brain connectivity states. Until now, how functional connectivity (FC) of the brain network changes with altitudes is still unclear. In this article, we used EEG data of the Go/NoGo paradigm from Weinan (347 m) and Nyingchi (2950 m). A combination of dynamic FC (dFC) and the K-means cluster was employed to extract dynamic FC states which were later distinguished by graph metrics. Besides, temporal properties of networks such as fractional windows (FW), transition numbers (TN) and mean dwell time (MDT) were calculated. Finally, we successfully extracted two different states from dFC matrices where State 1 was verified to have higher functional integration and segregation. The dFC states dynamically switched during the Go/NoGo tasks and the FW of State 1 showed a rise in the high-altitude participants. Also, in the regional analysis, we found higher state deviation in the fronto-parietal cortices and enhanced FC strength in the occipital lobe. These results demonstrated that long-term exposure to the high-altitude environment could lead brain networks to reorganize as networks with higher inter- and intra-networks information transfer efficiency, which could be attributed to a compensatory mechanism to the compromised brain function due to the plateau environment. This study provides a new perspective in considering how the plateau impacted cognitive impairment.
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BACKGROUND: Sepsis is a significant contributor to both intensive care unit (ICU) admissions and mortality among patients in ICU, with a rising prevalence of obesity. There is a lack of extensive research on the correlation between TyGI and findings in patients with sepsis, especially in obese patients. METHODS: This study used a retrospective cohort design and included patients with sepsis (≥18 years) from the Medical Information Mart for Intensive Care IV database. The association between TyGI and outcome was examined using multivariable logistic regression analysis. RESULTS: 8,840 patients with sepsis were included in the analysis. The in-ICU mortality rate was 9.7%. Non-survivors exhibited significantly greater TyGI levels than survivors [9.19(8.76-9.71) vs. 9.10(8.67-9.54), p < 0.001]. The adjusted multivariate regression model showed that elevated TyGI values were linked to a greater likelihood of death in ICU (odds ratio [OR] range 1.072-1.793, p < 0.001) and hospital (OR range 1.068-1.445, p = 0.005). Restricted Cubic Spline analysis revealed a nonlinear association between TyGI and in-ICU and in-hospital mortality risks within specified ranges. Subgroup analysis revealed interaction effects in the general obesity, abdominal obesity, and impaired fasting glucose subgroups (p = 0.014, 0.016, and < 0.001, respectively). CONCLUSION: TyGI was associated with an increased sepsis-related short-term mortality risk and adverse outcomes after ICU admission.
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Glicemia , Mortalidade Hospitalar , Unidades de Terapia Intensiva , Obesidade , Sepse , Triglicerídeos , Humanos , Sepse/mortalidade , Sepse/metabolismo , Estudos Retrospectivos , Masculino , Feminino , Pessoa de Meia-Idade , Obesidade/mortalidade , Obesidade/metabolismo , Obesidade/complicações , Idoso , Triglicerídeos/sangue , Triglicerídeos/metabolismo , Glicemia/análise , Glicemia/metabolismo , AdultoRESUMO
AIMS: There is growing evidence that the gut microbiota plays a significant part in the pathophysiology of chronic stress. The dysbiosis of the gut microbiota closely relates to dysregulation of microbiota-host cometabolism. Composition changes in the gut microbiota related to perturbations in metabolic profiles are vital risk factors for disease development. Hyperbaric oxygen therapy is commonly applied as an alternative or primary therapy for various diseases. Therefore, a metabolic and gut bacteria perspective is essential to uncover possible mechanisms of chronic stress and the therapeutic effect of hyperbaric oxygenation. We determined that there were significantly disturbed metabolites and disordered gut microbiota between control and chronic stress group. The study aims to offer further information on the interactions between host metabolism, gut microbiota, and chronic stress. METHODS: At present, chronic unpredictable mild stress is considered the most widespread method of modeling chronic stress in animals, so we used a chronic unpredictable mild stress mouse model to characterize changes in the metabolome and microbiome of depressed mice by combining 16S rRNA gene sequencing and UHPLC-MS/MS-based metabolomics. Pearson's correlation-based clustering analysis was performed with above metabolomics and fecal microbiome data to determine gut microbiota-associated metabolites. RESULTS: We found that 18 metabolites showed a significant correlation with campylobacterota. Campylobacterota associated metabolites were significantly enriched mainly in the d-glutamate and d-glutamine metabolism. Hyperoxia treatment may improve depression-like behaviors in chronic stress model mice through regulating the disrupted metabolites. CONCLUSIONS: Hyperbaric oxygen improves depression-like behaviors in chronic stress model mice by remodeling Campylobacterota associated metabolites.
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Microbioma Gastrointestinal , Oxigenoterapia Hiperbárica , Camundongos , Animais , Depressão/terapia , Depressão/metabolismo , RNA Ribossômico 16S/genética , Espectrometria de Massas em TandemRESUMO
Background: The choice of postoperative weight bearing protocol after uncemented total hip arthroplasty (THA) remains controversial. The aim of this study was to assess the efficacy and safety of immediate unrestricted weight bearing (UWB) compared with partial weight bearing (PWB) in patients undergoing uncemented THA. Methods: Relevant articles were retrieved from electronic databases. Both randomized controlled trials (RCTs) and non-RCTs were included but analyzed separately. All functional and clinical outcomes with at least 2 independent study outcomes were meta-analyzed. Results: A total of 17 studies were investigated. No adverse effect was found regarding micromotion of the femoral stem with immediate UWB following uncemented THA. There was also no correlation between immediate UWB and failure of ingrowth fixation and higher risks of femoral stem subsidence and surgical revision in RCTs. Harris hip score was better in patients with immediate UWB than those with PWB at 1 year post surgery, but the difference was not statistically significant. Conclusions: Immediate UWB did not have extra harm compared with PWB in patients undergoing uncemented THA. UWB was not superior to PWB. Considering the improvement of Harris score and the compliance of patients, UWB can be encouraged in THA rehabilitation.
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Chronic and recurrent inflammatory disorders of the gastrointestinal tract caused by a complex interplay between genetics and intestinal dysbiosis are called inflammatory bowel disease. As a result of the interaction between the liver and the gut microbiota, bile acids are an atypical class of steroids produced in mammals and traditionally known for their function in food absorption. With the development of genomics and metabolomics, more and more data suggest that the pathophysiological mechanisms of inflammatory bowel disease are regulated by bile acids and their receptors. Bile acids operate as signalling molecules by activating a variety of bile acid receptors that impact intestinal flora, epithelial barrier function, and intestinal immunology. Inflammatory bowel disease can be treated in new ways by using these potential molecules. This paper mainly discusses the increasing function of bile acids and their receptors in inflammatory bowel disease and their prospective therapeutic applications. In addition, we explore bile acid metabolism and the interaction of bile acids and the gut microbiota.
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Microbioma Gastrointestinal , Doenças Inflamatórias Intestinais , Animais , Humanos , Ácidos e Sais Biliares , Intestinos , Fígado , Doenças Inflamatórias Intestinais/tratamento farmacológico , Disbiose , MamíferosRESUMO
OBJECTIVE: To observe the effect of feeding on different pig tissues on the development of Chrysomya megacephala larvae. METHODS: About 200 larvae each were reared on four different substrates, i.e. pig's brain, liver, muscle and a mixture of minced pork muscle and fat (7 : 3) at a constant temperature of 25 degrees C. Length and weight of larvae and pupae were measured at 12 h interval 16 h after eclosion. 10 larvae or pupae were collected each time. The time of development, mortality, and sex ratio of adults were recorded. RESULTS: Three replicated experiments showed that the larvae fed on liver grew slowly, time of reaching maximum length and weight was delayed for about 24-36 h, and the duration of larva development was longer than that of other groups (P<0.01). The mean maximal larval length in mixture group [(14.89 +/- 0.39) mm] was statistically shorter than that of brain group, muscle group and liver group, [(17.81 +/- 0.54), (16.94 +/- 0.43) and (17.14 +/- 0.27) mm, respectively] (P<0.01). The mean maximal larval weight in liver group [(73.5 +/- 6.8) mg] and mixture group [(63.0 +/- 5.4) mg] was statistically lighter than brain group [(91.2 +/- 7.5) mg] and muscle group [(86.3 +/- 7.3) mg] (P<0.01). The pupal length in mixture group was statistically shorter than that of other 3 groups (P<0.01). The pupal weight of mixture group and liver group was statistically lighter than that of brain group and muscle group (P<0.01). The larval and pupal mortality of mixture group [(9.8 +/- 3.1)% and (8.9 +/- 3.1)%] was statistically higher than that of brain group [(5.5 +/- 3.1)% and (4.6 +/- 1.5)%], muscle group [(4.7 +/- 2.2)% and (3.8 +/- 2.0)%] and liver group [(5.4 +/- 2.3)% and (4.8 +/- 1.7)%] (P<0.01). There was no significant difference in the sex ratio among the four groups (P>0.05). CONCLUSION: The development duration of the larvae fed on liver is longer than other groups. The body length and weight of larvae and pupae fed on mixture diet are less than other groups with higher mortality.
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Dípteros/crescimento & desenvolvimento , Ingestão de Alimentos , Animais , Encéfalo , Larva/crescimento & desenvolvimento , Fígado , Carne , Músculos , SuínosRESUMO
OBJECTIVE: To research the pattern of larvae and pupae development when exposed to ketamine. METHODS: The larvae of Chrysomya megacephala were reared in artificial diet containing ketamine with concentration of 0, 25, 50 and 100 mg/kg respectively at 32 degrees C, 28 degrees C and 24 degrees C in environmental chamber with a 12-h photoperiod and 75% humidity. 10 samples were collected from each group every 12 h from the 16th h after hatching to pupation. The max body length, weight of the larvae, growth rate of body length, weight and developmental duration of each stage were observed. RESULTS: The average length and weight in the treatment groups were significantly less than the control before achieving the maximum (P < 0.05), and the growth rate of 1/2LD50 group at 24 degrees C was most retarded. No dose dependence were observed among the ketamine fed groups. The effect of ketamine dose, temperature and their interaction on larval length and weight was statistically significant (P < 0.01). The effect of ketamine dose, temperature and their interaction account for, respectively, 20.9%, 60.2% and 18.9% of the total effect on growth of larval length, and they account for 8.3%, 85.6% and 6.1% of the total effect on growth of larval weight. The duration of larval stage in treatment groups was significantly delayed in comparison to the control at different temperatures (P < 0.05), and the duration of prepupal stage in treatment groups was significantly delayed (P < 0.05). However, the duration of pupal stage in treatment groups at 24 degrees C was significantly shorter than the control (P < 0.05). CONCLUSION: The time achieving maximum length and weight was significantly delayed, which results in an increased development duration of larval and prepupal stages, indicating that ketamine inhibits the growth of the larvae of C. megacephala.
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Dípteros/efeitos dos fármacos , Dípteros/crescimento & desenvolvimento , Toxicologia Forense/métodos , Ketamina/farmacologia , Animais , Larva/efeitos dos fármacos , Larva/crescimento & desenvolvimento , Dose Letal Mediana , TemperaturaRESUMO
Background: Glycogen synthase kinase 3ß (GSK3B) is reported to be a protective factor for the degradation of chondrocytes by extracellular mechanisms. Nuclear receptor subfamily 4 group A member 3 (NR4A3) is a proinflammatory factor in osteoarthritis. Their regulation mechanism in posttraumatic osteoarthritis (PTOA) is not fully understood. Methods: GSK3B expression in the cartilage tissue of PTOA patients was analyzed by western blotting. IL-1ß-induced chondrocytes were transfected with pcDNA-GSK3B, and then, the cell viability, apoptosis, expression of the chondrocyte extracellular matrix degradation-related genes MMP13, aggrecan, and type II collagen, and secretion of inflammatory factors TNF-α and IL-6 were detected. Co-IP was used to analyze the interaction between GSK3B and DNMT1. Ch-IP and methylation-specific PCR assays were used for methylation. Also, cells were transfected with pcDNA-GSK3B or together with pcDNA-NR4A3, as well as transfected with si-NR4A3, and then, cell functions were tested. Then, the mice subjected to destabilization of medial meniscus (DMM) surgery were intra-articular injected with 100 µL of the following adeno-related virus vectors (empty vector, Ad-GSK3B, scrambled shRNA, and sh-NR4A3), respectively, and the virus titer was 2 × 108 TU/mL. Cartilage integrity was evaluated by safranin O/fast green staining, HE staining, and Osteoarthritis Research Society International (OARSI) score. Results: The expression of GSK3B protein was downregulated in PTOA patients. GSK3B overexpression alleviated IL-1ß-induced chondrocyte apoptosis and extracellular matrix degradation, as well as cartilage mineralization in PTOA model mice. NR4A3 overexpression reversed the effect of GSK3B on IL-1ß-induced chondrocyte functions. GSK3B could recruit DNMT1 to the NR4A3 promoter region to promote the methylation of NR4A3 and inhibit the expression of NR4A3 protein. Similarly, NR4A3 interference alleviated cartilage degradation under stimulating conditions by inhibiting the activation of the JAK2/STAT3 signaling pathway. Conclusion: GSK3B recruits DNMT1 to the NR4A3 promoter region and inhibits the activation of the NR4A3-mediated JAK2/STAT3 signaling pathway, thereby alleviating PTOA.
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Osteoartrite , Receptores de Esteroides , Animais , Células Cultivadas , Condrócitos/metabolismo , Colágeno Tipo II/genética , Colágeno Tipo II/metabolismo , Colágeno Tipo II/farmacologia , Proteínas de Ligação a DNA/genética , Glicogênio Sintase Quinase 3 beta/genética , Glicogênio Sintase Quinase 3 beta/metabolismo , Humanos , Interleucina-1beta/genética , Camundongos , Proteínas do Tecido Nervoso/genética , Osteoartrite/genética , Osteoartrite/metabolismo , Regiões Promotoras Genéticas , Receptores de Esteroides/genética , Receptores de Esteroides/metabolismo , Receptores dos Hormônios Tireóideos/genética , Receptores dos Hormônios Tireóideos/metabolismoRESUMO
Acute lung injury (ALI) is a common complication in patients with sepsis and is accompanied by high mortality. The present study aimed to investigate if the organic compound citrulline has a protective against lipopolysaccharide (LPS)-stimulated ALI and its potential mechanisms. ALI was induced in mice by intraperitoneal (i.p.) injection of LPS (10 mg/kg). Citrulline (1 g/kg/day) was administrated i.p. 7 days prior to LPS injection. Mouse lung vascular endothelial cells (MLVECs) were divided into five groups: Control, LPS, LPS + Cit, LPS + N-acetyl-L-cysteine (NAC) and LPS + Cit + ML385. Lung injury was determined by morphology changes. Apoptosis and pyroptosis were detected using western blot analysis and immunofluorescence. The present results indicated that citrulline can significantly attenuate ALI. Citrulline pretreatment decreased the expression of NOD-, LRR- and pyrin domain-containing protein 3 (NLRP3) inflammasome and decreased pyroptosis and apoptosis. Intervention with the total reactive oxygen species (ROS) scavenger N-acetyl-L-cysteine attenuated NLRP3 inï¬ammasome-associated pyroptosis and apoptosis in LPS-treated MLVECs. Citrulline pretreatment inhibited pyroptotic cell death and apoptosis induced by LPS. Citrulline decreased accumulation of intracellular ROS and activated the nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway. Furthermore, the Nrf2 inhibitor ML385 reversed ROS generation, NLRP3 inï¬ammasome-mediated pyroptosis and apoptosis suppressed by citrulline. In summary, the present data demonstrated that citrulline may confer protection against ALI via inhibition of ROS/NLRP3 inflammasome-dependent pyroptosis and apoptosis via the Nrf2 signaling pathway.
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Developing high-performance non-noble bifunctional catalysts is pivotal for large-scale seawater electrolysis but remains a challenge. Here we report a sandwichlike NiCo(HPO4)2@Ni3N/NF (denoted by NiCoHPi@Ni3N/NF) catalyst. Vertical Ni3N nanosheet arrays are first grown and supported on nickel foam, and then a bimetallic NiCoHPi coating is decorated on Ni3N nanosheets by one-step electrodeposition. The hierarchical sandwich like structure offers a large surface area and plenty of catalytic active sites, and the coupling of interconnected Ni3N and NiCoHPi accelerates the electron transfer. Moreover, the surficial hydrogen phosphate ions contribute to a proper OH- absorption capacity due to the Lewis acid-base reaction. As a result, the NiCoHPi@Ni3N/NF catalyst exhibits good OER and HER activity, requiring overpotentials of 365 mV (for OER) and 174 mV (for HER) to deliver 100 mA cm-2 in the alkaline simulated seawater electrolyte. When assembled the NiCoHPi@Ni3N/NF catalyst as both the anode and cathode, it only needs 1.86 V to reach 100 mA cm-2 in alkaline simulated seawater electrolyte. This work may inspire the design and exploration of self-supported hierarchical composite electrocatalysts for hydrogen production from the electrolysis of seawater.
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Rationale: The lung-protective effects of dopamine and its role in the pathology of ventilator-induced lung injury (VILI) are emerging. However, the underlying mechanisms are still largely unknown. Objective: To investigate the contribution of dopamine receptor dysregulation in the pathogenesis of VILI and therapeutic potential of dopamine D1 receptor (DRD1) agonist in VILI. Methods: The role of dopamine receptors in mechanical stretch-induced endothelial barrier dysfunction and lung injury was studied in DRD1 knockout mice, in isolated mouse lung vascular endothelial cells (MLVECs), and in lung samples from patients who underwent pulmonary lobectomy with mechanical ventilation for different time periods. Measurements and Main Results: DRD1 was downregulated in both surgical patients and mice exposed to mechanical ventilation. Prophylactic administration of dopamine or DRD1 agonist attenuated mechanical stretch-induced lung endothelial barrier dysfunction and lung injury. By contrast, pulmonary knockdown or global knockout of DRD1 exacerbated these effects. Prophylactic administration of dopamine attenuated mechanical stretch-induced α-tubulin deacetylation and subsequent endothelial hyperpermeability through DRD1 signaling. We identified that cyclic stretch-induced glycogen-synthase-kinase-3ß activation led to phosphorylation and activation of histone deacetylase 6 (HDAC6), which resulted in deacetylation of α-tubulin. Upon activation, DRD1 signaling attenuated mechanical stretch-induced α-tubulin deacetylation and subsequent lung endothelial barrier dysfunction through cAMP/exchange protein activated by cAMP (EPAC)-mediated inactivation of HDAC6. Conclusions: This work identifies a novel protective role for DRD1 against mechanical stretch-induced lung endothelial barrier dysfunction and lung injury. Further study of the mechanisms involving DRD1 in the regulation of microtubule stability and interference with DRD1/cAMP/EPAC/HDAC6 signaling may provide insight into therapeutic approaches for VILI.
Assuntos
Regulação para Baixo/fisiologia , Células Endoteliais/metabolismo , Endotélio Vascular/metabolismo , Pulmão/metabolismo , Receptores de Dopamina D1/metabolismo , Lesão Pulmonar Induzida por Ventilação Mecânica/metabolismo , Animais , AMP Cíclico/metabolismo , Desacetilase 6 de Histona/metabolismo , Humanos , Camundongos , Camundongos Knockout , Respiração Artificial/métodos , Transdução de Sinais/fisiologia , Estresse Mecânico , Tubulina (Proteína)/metabolismoRESUMO
Ventilatorassociated lung injury (VALI) remains a significant medical problem in intensive care units. The present study aimed to investigate the role of sphingosine kinase 1 (SPHK1) in VALI using a twohit model and explore the potential underlying molecular mechanism. Mice were divided into five groups: i) Nonventilated group; ii) nonventilated + lipopolysaccharide (LPS) group; iii) ventilated group; iv) ventilated + LPS group; and v) ventilated + LPS + SPHK1 inhibitor group. Mice were administered LPS (1 mg/kg) via an intraperitoneal injection. After 12 h, the mice were anesthetized and connected to a ventilator (10 ml/kg at 150 breaths/min) for 4 h. SPHK1 inhibitor (50 mg/kg) was injected intraperitoneally 1 h prior to ventilation. Mouse lung vascular endothelial cells were treated with LPS and SPHK1 inhibitor, and then subjected to cyclic stretch for 4 h. The present results suggested that the expression of SPHK1 and sphingosine 1 phosphate was upregulated in the twohit model of VALI; SPHK1 inhibitor could attenuate VALI in the twohit model as observed by hematoxylin and eosin staining, and affected the cell count and the protein content levels in the bronchoalveolar lavage fluid. In addition, treatment with SPHK1 inhibitor reduced the wettodry ratio of the lungs and suppressed Evans blue dye leakage into the lung tissue. Furthermore, SPHK1 inhibitor exhibited protective effects on the twohit model of VALI by inhibiting the Ras homolog family member amediated phosphorylation of myosin phosphatase target subunit 1 (MYPT1) and endothelial hyperpermeability. Additionally, mice were divided into five additional groups: i) Nonventilated group; ii) nonventilated + LPS group; iii) ventilated group; iv) ventilated + LPS group; and v) ventilated + LPS + Rhoassociated coiledcoil forming protein kinase (ROCK)1 inhibitor group. ROCK1 inhibitor (10 mg/kg) was injected intraperitoneally 1 h prior to ventilation. The present results suggested that ROCK1 inhibitor could attenuate mechanical stretchinduced lung endothelial injury and the phosphorylation of MYPT1 in vivo and in vitro. Collectively, the present findings indicated that upregulation of SPHK1 may contribute to VALI in a twohit model.
Assuntos
Fosfatase de Miosina-de-Cadeia-Leve/genética , Fosfotransferases (Aceptor do Grupo Álcool)/genética , Lesão Pulmonar Induzida por Ventilação Mecânica/genética , Quinases Associadas a rho/genética , Animais , Líquido da Lavagem Broncoalveolar/química , Permeabilidade Capilar/efeitos dos fármacos , Modelos Animais de Doenças , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/patologia , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Lipopolissacarídeos/toxicidade , Lisofosfolipídeos/metabolismo , Camundongos , Fosforilação , Fosfotransferases (Aceptor do Grupo Álcool)/antagonistas & inibidores , Transdução de Sinais/efeitos dos fármacos , Esfingosina/análogos & derivados , Esfingosina/metabolismo , Lesão Pulmonar Induzida por Ventilação Mecânica/induzido quimicamente , Lesão Pulmonar Induzida por Ventilação Mecânica/tratamento farmacológico , Lesão Pulmonar Induzida por Ventilação Mecânica/patologia , Quinases Associadas a rho/antagonistas & inibidoresRESUMO
High-mobility group box 1 (HMGB1) contributes to lung vascular hyperpermeability during ventilator-induced lung injury. We aimed to determine whether the natural antioxidant resveratrol protected against HMGB1-induced endothelial hyperpermeability both in vitro and in vivo. We found that HMGB1 decreased vascular endothelial (VE)-cadherin expression and increased endothelial permeability, leading to mitochondrial oxidative damage in primary cultured mouse lung vascular endothelial cells (MLVECs). Both the mitochondrial superoxide dismutase 2 mimetic MnTBAP and resveratrol blocked HMGB1-induced mitochondrial oxidative damage, VE-cadherin downregulation, and endothelial hyperpermeability. In in vivo studies, anesthetized male ICR mice were ventilated for 4h using low tidal volume (6 ml/kg) or high tidal volume (HVT; 30 ml/kg) ventilation. The mice were injected intraperitoneally with resveratrol immediately before the onset of ventilation. We found that resveratrol attenuated HVT-associated lung vascular hyperpermeability and HMGB1 production. HVT caused a significant increase in nuclear factor-erythroid 2-related factor 2 (Nrf2) nuclear translocation and Nrf2 target gene expression in lung tissues, which was further enhanced by resveratrol treatment. HMGB1 had no effect on Nrf2 activation, whereas resveratrol treatment activated the Nrf2 signaling pathway in HMGB1-treated MLVECs. Moreover, Nrf2 knockdown reversed the inhibitory effects of resveratrol on HMGB1-induced mitochondrial oxidative damage and endothelial hyperpermeability. The inhibitory effect of resveratrol on cyclic stretch-induced HMGB1 mRNA expression in primary cultured MLVECs was also abolished by Nrf2 knockdown. In summary, this study demonstrates that resveratrol protects against lung endothelial barrier dysfunction initiated by HVT. Lung endothelial barrier protection by resveratrol involves inhibition of mechanical stretch-induced HMGB1 release and HMGB1-induced mitochondrial oxidative damage. These protective effects of resveratrol might be mediated through an Nrf2-dependent mechanism.
Assuntos
Antioxidantes/farmacologia , Proteína HMGB1/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Estilbenos/farmacologia , Lesão Pulmonar Induzida por Ventilação Mecânica/metabolismo , Animais , Western Blotting , Modelos Animais de Doenças , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/metabolismo , Ensaio de Imunoadsorção Enzimática , Citometria de Fluxo , Imunofluorescência , Técnicas de Silenciamento de Genes , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Pulmão/patologia , Masculino , Camundongos , Camundongos Endogâmicos ICR , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Mitocôndrias/patologia , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/fisiologia , RNA Interferente Pequeno , Reação em Cadeia da Polimerase em Tempo Real , Resveratrol , Estresse Mecânico , Transfecção , Lesão Pulmonar Induzida por Ventilação Mecânica/fisiopatologiaRESUMO
Objective: To screen and analysis of antihepatic steatosis active components and cell metabolism withinrhubarb anthraquinones in vitro. Methods: Lipid-lowering activity of liver cell affinity components in rhubarbanthraquinones was evaluated by hepatic steatosis in vitro model. Hepatocyte-specific affinity compositions werescreened by hepatocyte affinity chromatography. Hep G2 cells were incubated with rhubarb anthraquinones in vitro andwere detected by HPLC and HPLC-Q-TOF-MS method. Results: Compared with normal group, the intracellular TGcontent of model group were significantly increased (P<0.05) . Respectively treated with different concentrations ofrhubarb anthraquinones, the intracellular TG content were significantly decreased in a dose dependent manner (P<0.05) in comparison with model group. Compared with rhubarb solution, there are mainly four hepatocyte-specific affinitycompositions of aloe-emodin, emodin, chrysophanol and physcion, except for rhein. Conclusion: The lipid-lowering experiment in vitro verified that five rhubarb anthraquinones have significant lipid-lowering activity. This research established a biochromatography method to screen out four hepatocyte-specific affinity compositions of aloe-emodin, emodin, chrysophanol and physcion from rhubarb anthraquinones, while the conjugation sites of rhein were ambiguous.Combined hepatocyte affinity chromatography with hepatic steatosis in vitro model, it could evaluate and screen lipidlowering active ingredient in vitro and cell metabolism of Chinese traditional medicine for the drug development of lipidlowering rhubarb anthraquinones.