RESUMO
Recent evidence suggests that glia maturation factor ß (GMFß) is important in the pathogenesis of pulmonary arterial hpertension (PAH), but the underlying mechanism is unknown. To clarify whether GMFß can be involved in pulmonary vascular remodeling and to explore the role of the IL-6-STAT3 pathway in this process, the expression of GMFß in PAH rats is examined and the expression of downstream molecules including periostin (POSTN) and interleukin-6 (IL-6) is measured using real-time quantitative polymerase chain reaction (RT-qPCR) and western blot analysis. The location and expression of POSTN is also tested in PAH rats using immunofluorescence. It is proved that GMFß is upregulated in the lungs of PAH rats. Knockout GMFß alleviated the MCT-PAH by reducing right ventricular systolic pressure (RVSP), mean pulmonary arterial pressure (mPAP), and pulmonary vascular remodeling. Moreover, the inflammation of the pulmonary vasculature is ameliorated in PAH rats with GMFß absent. In addition, the IL-6-STAT3 signaling pathway is activated in PAH; knockout GMFß reduced POSTN and IL-6 production by inhibiting the IL-6-STAT3 signaling pathway. Taken together, these findings suggest that knockout GMFß ameliorates PAH in rats by inhibiting the IL-6-STAT3 signaling pathway.
Assuntos
Fator de Maturação da Glia , Interleucina-6 , Remodelação Vascular , Animais , Remodelação Vascular/genética , Remodelação Vascular/fisiologia , Ratos , Masculino , Interleucina-6/metabolismo , Interleucina-6/genética , Fator de Maturação da Glia/metabolismo , Fator de Maturação da Glia/genética , Hipertensão Arterial Pulmonar/metabolismo , Hipertensão Arterial Pulmonar/fisiopatologia , Hipertensão Arterial Pulmonar/genética , Hipertensão Arterial Pulmonar/patologia , Transdução de Sinais , Ratos Sprague-Dawley , Fator de Transcrição STAT3/metabolismo , Fator de Transcrição STAT3/genética , Moléculas de Adesão Celular/metabolismo , Moléculas de Adesão Celular/genética , Artéria Pulmonar/metabolismo , Artéria Pulmonar/patologia , Artéria Pulmonar/fisiopatologia , Modelos Animais de DoençasRESUMO
Background: The effects of glycemic status on coronary physiology have not been well evaluated. This study aimed to investigate changes in coronary physiology by using angiographic quantitative flow ratio (QFR), and their relationships with diabetes mellitus (DM) and glycemic control status. Methods: This retrospective cohort study included 530 patients who underwent serial coronary angiography (CAG) measurements between January 2016 and December 2021 at Tongji Hospital of Tongji University. Based on baseline and follow-up angiograms, 3-vessel QFR (3V-QFR) measurements were performed. Functional progression of coronary artery disease (CAD) was defined as a change in 3V-QFR (Δ3V-QFR = 3V-QFRfollow-up - 3V-QFRbaseline) ≤-0.05. Univariable and multivariable logistic regression analyses were applied to identify the independent predictors of coronary functional progression. Subgroup analysis according to diabetic status was performed. Results: During a median interval of 12.1 (10.6, 14.3) months between the two QFR measurements, functional progression was observed in 169 (31.9%) patients. Follow-up glycosylated hemoglobin (HbA1c) was predictive of coronary functional progression with an area under the curve (AUC) of 0.599 [95% confidence interval (CI): 0.546-0.651; P<0.001] in the entire population. Additionally, the Δ3V-QFR values were significantly lower in diabetic patients with HbA1c ≥7.0% compared to those with well-controlled HbA1c or non-diabetic patients [-0.03 (-0.09, 0) vs. -0.02 (-0.05, 0.01) vs. -0.02 (-0.05, 0.02); P=0.002]. In a fully adjusted multivariable logistics analysis, higher follow-up HbA1c levels were independently associated with progression in 3V-QFR [odds ratio (OR), 1.263; 95% CI: 1.078-1.479; P=0.004]. Furthermore, this association was particularly strong in diabetic patients (OR, 1.353; 95% CI: 1.082-1.693; P=0.008) compared to patients without DM. Conclusions: Among patients with established CAD, on-treatment HbA1c levels were independently associated with progression in physiological atherosclerotic burden, especially in patients with DM.
RESUMO
Background: Surgery for acetabular fractures involving both columns is difficult and traumatic, making it necessary to explore a minimally invasive and accurate surgical method. Methods: This retrospective case-control study analyzed the clinical data of 34 patients and divided them into two groups: a control group (9 males and 8 females) and a research group (11 males and 6 females) with acetabular fractures involving the anterior and posterior columns. All patients were placed in the supine position via the pararectus approach. A three-dimensional (3D) guide was placed at the position where the posterior column screw was inserted in the second window, and a posterior column screw was placed percutaneously on the medial side of the iliac spine in the research group. The operation time, intraoperative blood loss, and fracture union time of the two groups were recorded. Pelvic radiographs and computed tomography (CT) scans were routinely performed before and after surgery to evaluate reduction and fixation. Residual gap and step displacement were measured using a standardized CT-based method after the surgery. Hip mobility was assessed according to the modified Merle, d'Aubigné, and Postel criteria. Results: All patients were followed up for 6-30 (16.941±6.571) months. The operation times of the two groups were 126 [interquartile range (IQR), 95-133] min (control group) and 110 (IQR, 85-124) min (research group), the intraoperative blood losses were 430 (IQR, 290-550) mL (control group) and 380 (IQR, 260-500) mL (research group). All patients achieved bone healing, with a union time of 15 (IQR, 12-17) weeks (control group) and 13 (IQR, 11.5-15) weeks (research group). According to the standardized CT-based method, the reduction after surgery was acceptable in 13 (control group) and 14 (research group) of these patients (defined as a gap <5 mm or a step-off <1 mm), and the anatomical reduction rates were 76.47% and 82.35%, respectively. Conclusions: The use of a single pararectus approach combined with 3D guide-assisted percutaneous anterograde posterior column screws can shorten the operation time and place effective posterior column screws precisely with minimal invasiveness. At the same time, the acetabular reduction and functional recovery are satisfactory, and there are fewer postoperative complications, which makes this procedure an ideal surgical option.
RESUMO
Objectives: Mendelian randomization (MR) was used to estimate the causal relationship between body mass index (BMI), ever smoked, heart failure, alcohol intake frequency, inflammatory bowel disease (IBD), and pulmonary embolism (PE). This study aimed to investigate whether there is a causal relationship between BMI, the presence of smoking, heart failure, frequency of alcohol intake, IBD, and PE. Methods: Pooled data on PE from a published GWAS meta-analysis involving approximately 461,164 participants of European ancestry were selected. A publicly available pooled dataset of BMI (461,460), ever smokers (461,066), heart failure (977,323), IBD (75,000), and frequency of alcohol intake (462,346) was used from another independent GWAS. MR was performed using established analysis methods, including Wald ratios, inverse variance weighted (IVW), weighted median (WM), and MR-Egger. Also, the final expansion was validated with multivariate MR. Results: In the IVW model, genetically elevated BMI was causally associated with PE [OR = 1.002, 95% CI (1.001, 1004), P = 0.039]. Cochran's Q test was used to detect heterogeneity in the MR-Egger analysis (P = 0.576). Therefore, the effect of gene-level heterogeneity was not considered. In the MR analysis of other risk factors, we observed genes for ever smoking [IVW OR = 1.004, 95% CI (0.997, 1.012)], heart failure [IVW OR = 0.999, 95% CI (0.996, 1.001)], IBD [IVW OR = 1.000, 95% CI (0.999, 1.001)], and frequency of alcohol intake [IVW OR = 1.002, 95% CI (1.000, 1.004)] were not causally associated with PE. Analysis using multivariate MR expansion showed no causal effect of BMI on PE considering the effect of height as well as weight (P = 0.926). Conclusion: In European populations, a causal relationship exists between BMI and PE: increased BMI leads to PE. In contrast, ever smoking, heart failure, frequency of alcohol intake, and IBD are not directly associated with PE. There was no causal effect of BMI with PE in multivariate Mendelian randomized analysis.
RESUMO
Chronic heart failure (CHF) is a prevalent health concern with complex pathogenesis. This current study set out to estimate the function of the miR-129-5p/Smurf1/PTEN axis on cardiac function injury in CHF. The model of CHF in rats was established. The cardiac function indexes, myocardial tissue damage, and oxidative stress-related factors in CHF rats were evaluated after the interference of Smurf1/miR-129-5p/PTEN. The targeting relationships between miR-129-5p and Smurf1 and between PTEN and Smurf1 were verified. It was found that that after modeling, cardiac functions were impaired, heart/left ventricular/lung weight and the myocardial structure was destroyed, and the degree of fibrosis of myocardial tissue was increased. After Smurf1 knockdown, the cardiac function, myocardial structure, and oxidative stress were improved, and the fibrosis in myocardial tissue was decreased. Smurf1 was a target of miR-129-5p. miR-129-5p could annul the protective effect of Smurf1 silencing on CHF rats. Smurf1 inhibited PTEN expression by promoting PTEN ubiquitination, while miR-129-5p enhanced PTEN expression by inhibiting Smurf1. Meanwhile, overexpression of PTEN annulled the cardiac dysfunction in CHF rats induced by Smurf1. In conclusion, miR-129-5p targeted Smurf1 and repressed the ubiquitination of PTEN, and promoted PTEN expression, thus improving the cardiac function of CHF rats.
Assuntos
Regulação Enzimológica da Expressão Gênica , Insuficiência Cardíaca/metabolismo , MicroRNAs/metabolismo , PTEN Fosfo-Hidrolase/biossíntese , Ubiquitina-Proteína Ligases/metabolismo , Animais , Doença Crônica , Insuficiência Cardíaca/genética , Masculino , MicroRNAs/genética , PTEN Fosfo-Hidrolase/genética , Ratos , Ratos Wistar , Ubiquitina-Proteína Ligases/genéticaRESUMO
BACKGROUND: To investigate the protective efficacy of miR-155 on down regulating NADPH oxidase isoform subunit A1 (NoxA1) gene expression, resulting in inhibition of VSMC migration and over proliferation and thus ameliorating the progression of arterial atherosclerosis in AS mouse model. Therefore, to further explore the regulatory effect of miR-155 on neointima formation in AS and locate potential anti-atherosclerosis target. METHODS: The mouse vascular aorta smooth muscle cell (MOVAS) was cultured and transfected with recombinant Pad2YFG adenovirus fluorescent vector with miR-155 fragment into 4 groups. Western blotting and RT-PCR were performed to identify the expression of NoxA1 under different circumstances. Fluorescence microscope was applied to observe the transfection rate of miR-155 into adenovirus. Twelve-week fatty food induced atherosclerotic ApoE-/- mouse model was established as host to accept miR-155 transfected adenovirus transplantation to observe its effect on VSMC in AS progression. Carotid and thoracic artery were extracted at 1 month after dosing. Distribution of miR-155 was quantified via expression levels of protein and RNA to detect NoxA1, Nox1, p47phox and NADPH expression. Immunohistochemistry, fluorescence imaging and other methods were performed in arteries section to compare the thickness of neointima and assess the severity of AS in each group. RESULTS: Luciferase reporter gene assay showed significant expression of miR-155 in mimic group indicating that miR-155 had target binding effect with NoxA1 gene. Western blotting and RT-PCR results both showed significantly decreased NoxA1 expression in miR-155 mimic group while increased with its inhibitor. The miR-155 distribution was observed varied at 1 month after in control, miR-155 mimic and inhibitor groups. The NoxA1, NADPH, Nox1 and pp47phox protein expression in VSMC was decreased in mimic group vs control and inhibitor groups (P<0.05); no significant difference of NADPH expression was observed in all groups. The NoxA1, Nox1 and p47phox gene expression in VSMC were both found reduced compared with those of control group at week 4 (P<0.05). Immunohistochemistry staining of artery frozen sections figured out that the thickness of neointima of carotid artery in miR-155 mimic group was significantly lower vs control and inhibitor groups (P<0.01) at week 4. CONCLUSIONS: miR-155 played an important role in NoxA1-related signaling pathway. miR-155 transfection into VSMC may have anti-inflammatory regulatory effect on NoxA1 expression in vivo and resulting in amelioration of atherosclerotic lesion in AS mouse model. In summary, miR-155 specifically plays in a negative feedback loop and demonstrates a protective role during atherosclerosis-associated VSMC proliferation and neointima formation through the miR-155-NoxA1-p47phox complex signaling pathway.
RESUMO
Metaplasticity is referred to adjustment in the requirements for induction of synaptic plasticity based on the prior history of activity. Synaptic plasticity, including long-term potentiation (LTP) and long-term depression (LTD), has been considered to be the neural processes underlying learning and memory. Previous observations that cordycepin (an adenosine derivative) improved learning and memory seemed to be contradictory to the findings that cordycepin inhibited LTP. Therefore, we speculated that the conflicting reports of cordycepin might be related to metaplasticity. In the current study, population spike (PS) in hippocampal CA1 area of rats was recorded by using electrophysiological method in vivo. The results showed that cordycepin reduced PS amplitude in hippocampal CA1 with a concentration-dependent relationship, and high frequency stimulation (HFS) failed to induce LTP when cordycepin was intrahippocampally administrated but improved LTP magnitude when cordycepin was pre-treated. Cordycepin increased LTD induced by activating N-Methyl-D-aspartate (NMDA) receptors and subsequently facilitated LTP induced by HFS. Furthermore, we found that 1,3-dipropyl-8-cyclopentylxanthine (DPCPX), an adenosine A1 receptors antagonist, could block the roles of cordycepin on LTD and LTP. Collectively, cordycepin was able to modulate metaplasticity in hippocampal CA1 area of rats through adenosine A1 receptors. These findings would be helpful to reconcile the conflicting reports in the literatures and provided new insights into the mechanisms underlying cognitive function promotions with cordycepin treatment.
Assuntos
Agonistas do Receptor A1 de Adenosina/farmacologia , Região CA1 Hipocampal/efeitos dos fármacos , Desoxiadenosinas/farmacologia , Plasticidade Neuronal/efeitos dos fármacos , Receptor A1 de Adenosina/efeitos dos fármacos , Potenciais de Ação/efeitos dos fármacos , Animais , Região CA1 Hipocampal/metabolismo , Potenciação de Longa Duração/efeitos dos fármacos , Depressão Sináptica de Longo Prazo/efeitos dos fármacos , Masculino , Ratos Sprague-Dawley , Receptor A1 de Adenosina/metabolismo , Receptores de N-Metil-D-Aspartato/metabolismo , Fatores de TempoRESUMO
BACKGROUND: Telomerase plays an essential role in cancer cell proliferation. In this study, we investigated inhibition mechanism of aloe emodin (AE) on three different types of breast cancer cell lines, MDA-MB-453, MDA-MB-231 and MCF-7. METHODS: The cells were treated with different concentrations of AE. Relative length of telomere and human telomerase reverse-transcriptase (hTERT) mRNA level was analyzed by quantitative PCR (qPCR). Protein level was assayed by Western blot. Sodium bisulfite methylation sequencing was performed to assess the methylation status of gene promoter. Enzymology kinetics was applied to reveal the interaction between AE and telomerase. Ultraviolet-visible titration and fluorescence resonance energy transfer (FRET) melting experiment were carried out to study the interaction between AE and telomeric DNA. RESULTS: Continuous AE exposure of these cells for 48 h results in shortening of telomeres and inhibition of telomerase. The transcription of hTERT was repressed by activation of E2F1 and inactivation of c-myc proteins. Significant demethylation of CpG islands in hTERT gene promoter was observed in MDA-MB-453 and MCF-7 cells. AE competed with dNTP for occupation of the enzyme active site. AE was a telomeric G-quadruplex structure stabilizer as indicated by titration test and FRET experiments. CONCLUSIONS: AE was a competitive inhibitor of telomerase and a G-quadruplex structure stabilizer. AE decreased the transcription of hTERT gene in the three breast cancer cell lines via up-regulation E2F1 and down-regulation c-myc expressions. The suppressed transcription was also related to the demethylation of the gene promoter.
Assuntos
Antraquinonas/farmacologia , Neoplasias da Mama/tratamento farmacológico , Telomerase/metabolismo , Transcrição Gênica/efeitos dos fármacos , Neoplasias da Mama/metabolismo , Linhagem Celular Tumoral , Ilhas de CpG/efeitos dos fármacos , Regulação para Baixo/efeitos dos fármacos , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Células MCF-7 , Regiões Promotoras Genéticas/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-myc/metabolismo , RNA Mensageiro/metabolismo , Ativação Transcricional/efeitos dos fármacosRESUMO
Hepatoblastoma is a malignant tumor in the liver of children that generally occurs at the age of 2-3 years. There have been ample evidence from the preclinical as well as clinical studies suggesting the activation of Wnt/ß-catenin signaling in hepatoblastoma, which is mainly attributed to the somatic mutations in the exon 3 of ß-catenin gene. There is increased translocation of ß-catenin protein from the cell surface to cytoplasm and nucleus and intracellular accumulation is directly linked to the severity of the cancer. Accordingly, the alterations in ß-catenin and its target genes may be used as markers in the diagnosis and prognosis of pediatric live tumors. Furthermore, scientists have reported the therapeutic usefulness of inhibition of Wnt/ß-catenin signaling in hepatoblastoma and this inhibition of signaling has been done using different methods including short interfering RNA (siRNA), miRNA and pharmacological agents. Wnt/ß-catenin works in association with other signaling pathways to induce the development of hepatoblastoma including Yes-associated protein (YAP)1 (YAP-1), mammalian target of rapamycin (mTOR) 1 (mTOR-1), SLC38A1, glypican 3 (GPC3), nuclear factor κ-light-chain-enhancer of activated B cells (NF-kB), epidermal growth factor receptor, ERK1/2, tumor necrosis factor-α (TNF-α), regenerating islet-derived 1 and 3 α (REG1A and 3A), substance P (SP)/neurokinin-1 receptor and PARP-1. The present review describes the key role of Wnt/ß-catenin signaling in the development of hepatoblastoma. Moreover, the role of other signaling pathways in hepatoblastoma in association with Wnt/ß-catenin has also been described.
Assuntos
Hepatoblastoma/genética , Neoplasias Hepáticas/genética , Proteínas de Neoplasias/genética , beta Catenina/genética , Proliferação de Células/genética , Regulação Neoplásica da Expressão Gênica/genética , Hepatoblastoma/patologia , Humanos , Neoplasias Hepáticas/patologia , Via de Sinalização Wnt/genéticaRESUMO
Cordycepin, an adenosine analog, has been reported to improve cognitive function, but which seems to be inconsistent with the reports showing that cordycepin inhibited long-term potentiation (LTP). Behavioral-LTP is usually used to study long-term synaptic plasticity induced by learning tasks in freely moving animals. In order to investigate simultaneously the effects of cordycepin on LTP and behavior in rats, we applied the model of behavioral-LTP induced by Y-maze learning task through recording population spikes in hippocampal CA1 region. Golgi staining and Sholl analysis were employed to assess the morphological structure of dendrites in pyramidal cells of hippocampal CA1 area, and western blotting was used to examine the level of adenosine A1 receptors and A2A receptors (A2AR). We found that cordycepin significantly improved behavioral-LTP magnitude, accompanied by increases in the total length of dendrites, the number of intersections and spine density but did not affect Y-maze learning task. Furthermore, cordycepin obviously reduced A2AR level without altering adenosine A1 receptors level; and the agonist of A2AR (CGS 21680) rather than antagonist (SCH 58261) could reverse the potentiation of behavioral-LTP induced by cordycepin. These results suggested that cordycepin improved behavioral-LTP and morphological structure of dendrite in hippocampal CA1 but did not contribute to the improvement of learning and memory. And cordycepin improved behavioral-LTP may be through reducing the level of A2AR in hippocampus. Collectively, the effects of cordycepin on cognitive function and LTP were complex and involved multiple mechanisms.
Assuntos
Região CA1 Hipocampal/efeitos dos fármacos , Dendritos/efeitos dos fármacos , Desoxiadenosinas/farmacologia , Potenciação de Longa Duração/efeitos dos fármacos , Aprendizagem em Labirinto/efeitos dos fármacos , Animais , Masculino , Células Piramidais/efeitos dos fármacos , Ratos , Ratos Sprague-DawleyRESUMO
Sphingolipids have been accorded numerous biological functions however, the effects of feeding a western diet (diet rich in cholesterol and fat) on skin phenotypes, and color is not known. Here, we observed that chronic high-fat and high-cholesterol diet intake in a mouse model of atherosclerosis (ApoE-/-) decreases the level of ceramides and glucosylceramide. At the expense of increased levels of lactosylceramide due to an increase in the expression of lactosylceramide synthase (GalT-V). This is accompanied with neutrophil infiltration into dermis, and enrichment of tumor necrosis factor-stimulated gene-6 (TSG-6) protein. This causes skin inflammation, hair discoloration and loss, in ApoE-/- mice. Conversely, inhibition of glycosphingolipid synthesis, by D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (D-PDMP), unbound or encapsulated in a biodegradable polymer (BPD) reversed these phenotypes. Thus, inhibition of glycosphingolipid synthesis represents a unique therapeutic approach relevant to human skin and hair Biology.
Assuntos
Alopecia/patologia , Apolipoproteínas E/deficiência , Dieta Ocidental , Comportamento Alimentar , Glicoesfingolipídeos/biossíntese , Inflamação/patologia , Pele/patologia , Animais , Apolipoproteínas E/metabolismo , Moléculas de Adesão Celular/metabolismo , Ceramidas/metabolismo , Galactosiltransferases/metabolismo , Homeostase , Masculino , Camundongos , Modelos Biológicos , Morfolinas/farmacologia , Infiltração de Neutrófilos/efeitos dos fármacos , Neutrófilos/metabolismo , Fenótipo , Polímeros/farmacologiaRESUMO
A Disintegrin And Metalloproteinase 12 (ADAM12) is highly expressed in multiple cancers such as breast and cervical cancers and its high expression reduces the overall patient survival rate. ADAM12 has two major splicing variants, the long membrane-anchored form ADAM12L and the short secreted form ADAM12S. However, how they are regulated and whether they are modulated similarly or differently in cells are not clear. Here, we use affinity purification and mass spectrometry to identify the ADAM12S-interacting proteins. Spectral counting and MaxQuant label-free quantification reveal that ADAM12S but not ADAM12L specifically interacts with a subset of endoplasmic reticulum proteins, such as endoplasmin (GRP94), 78â¯kDa glucose-regulated protein (GRP78), and UDP-glucose:glycoprotein glucosyltransferase I (UGGT1), that regulate the folding and processing of secreted proteins. Further biochemical experiments validate the interaction between ADAM12S and several of its interacting proteins. Computational docking analysis demonstrates that GRP94 preferentially interacts with ADAM12S over ADAM12L. The data also suggest that both the protein expression level and the secretion of ADAM12S are regulated by GRP94 expression and knockdown. Our results reveal a link between these two proteins that are highly expressed in cancer cells. Furthermore, our studies define a new ADAM12S-specific regulator that may contribute to the cancer development. SIGNIFICANCE: A Disintegrin And Metalloproteinase 12 (ADAM12) is highly expressed in many cancers such as lung, breast, and cervical cancers. ADAM12 has two major splicing variants, the long membrane-anchored form ADAM12L and the short secreted form ADAM12S. However, how they are regulated and whether they are modulated similarly or differently are not completely understood. We use affinity purification and label-free quantitative proteomics to identify the ADAM12S-interacting proteins. Our results reveal that ADAM12S specifically interacts with a subset of endoplasmic reticulum proteins, including endoplasmin (GRP94), UDP-glucose:glycoprotein glucosyltransferase I (UGGT1), and neutral α-glucosidase AB (GANAB). Computer modeling reveals that ADAM12S interacts with the surface amino acids of GRP94 more strongly than ADAM12L. Biochemical experiments further reveal that GRP94 regulates both the protein level and the secretion of ADAM12S. Database mining finds that both GRP94 and ADAM12 are highly expressed in multiple cancers and their high expression is correlated with poor patient survival rate. Taken together, our work discovers a new upstream regulator for ADAM12S, which may contribute to its distinct functions in the regulation of the migration and invasion of cancer cells.
Assuntos
Proteína ADAM12/metabolismo , Glicoproteínas de Membrana/fisiologia , Proteômica/métodos , Linhagem Celular Tumoral , Cromatografia de Afinidade , Retículo Endoplasmático/química , Chaperona BiP do Retículo Endoplasmático , Proteínas de Choque Térmico HSP70 , Humanos , Espectrometria de Massas , Glicoproteínas de Membrana/metabolismo , Proteínas de Membrana , Simulação de Acoplamento Molecular , Neoplasias/etiologia , Ligação Proteica , Isoformas de ProteínasRESUMO
Peroxidase (POD) from jackfruit bulb was purified using ammonium sulfate precipitation, hydrophobic interaction and gel filtration columns. The POD was a dimer with a molecular weight of 104kDa. The Km and Vmax values for guaiacol, gallic acid and ophenylenediamine (OPD) were estimated. OPD was the most suitable substrate. The enzyme showed its maximum activity at pH5.5 and 55-60°C. The activation energy (Ea) of heat inactivation was estimated to be 206.40kJ/mol. The enthalpy, free energy and entropy values for the thermal inactivation were also determined. The POD activity was enhanced by K+, Zn2+, Ba2+, citric acid, malic acid, benzoic acid and EDTA·Na2, but inhibited by Cu2+, Ca2+, glutathione, cysteine and ascorbic acid. Chemical modification indicated a histidine residue was located in the enzyme active site. The POD activity in fruit extracts significantly decreased when heated at 80°C and 90°C. The ferric-reducing antioxidant power, ABTS radical scavenging activity and total phenolics decreased with increasing heating temperature and time.
Assuntos
Artocarpus/enzimologia , Peroxidases/isolamento & purificação , Proteínas de Plantas/isolamento & purificação , Antioxidantes/isolamento & purificação , Antioxidantes/farmacologia , Catecol Oxidase/antagonistas & inibidores , Catecol Oxidase/isolamento & purificação , Catecol Oxidase/farmacologia , Cátions/farmacologia , Inibidores Enzimáticos/farmacologia , Aditivos Alimentares/farmacologia , Sequestradores de Radicais Livres/isolamento & purificação , Sequestradores de Radicais Livres/farmacologia , Temperatura Alta , Concentração de Íons de Hidrogênio , Cinética , Peso Molecular , Oxirredução , Peroxidases/antagonistas & inibidores , Peroxidases/farmacologia , Fenóis/análise , Proteínas de Plantas/agonistas , Proteínas de Plantas/antagonistas & inibidores , Proteínas de Plantas/farmacologia , Raízes de Plantas/enzimologia , Estabilidade Proteica , Especificidade por SubstratoRESUMO
Immune activation and inflammation participate in the progression of chronic heart failure (CHF). Th17 cells and CD4+CD25+ regulatory T (Treg) cells both come from naive Th cells and share reciprocal development pathways but exhibit opposite effects. We hypothesized that the Th17/Treg balance was impaired in patients with CHF, and exercise can improve it. Rats with ischemic cardiomyopathy were prepared by ligaturing the left anterior descending branch of the left coronary artery. Rats in training group were trained with treadmill; Th17 cells increased significantly while Treg cells significantly decreased in s by flow cytometry, and the peripheral blood level of IL-6, IL-17, and TNF-α was obviously elevated by ELISA assay. We found that Th17/Treg balance is impaired in CHF rats, suggesting Th17/Treg imbalance potentially plays a role in the pathogenesis of CHF. Exercise can improve Th17/Treg imbalance, which also improves cardiac function of CHF.
Assuntos
Cardiomiopatias/terapia , Terapia por Exercício/métodos , Insuficiência Cardíaca/terapia , Isquemia Miocárdica/complicações , Linfócitos T Reguladores/imunologia , Células Th17/imunologia , Animais , Cardiomiopatias/sangue , Cardiomiopatias/imunologia , Cardiomiopatias/fisiopatologia , Modelos Animais de Doenças , Ecocardiografia , Insuficiência Cardíaca/sangue , Insuficiência Cardíaca/imunologia , Insuficiência Cardíaca/fisiopatologia , Interleucina-17/sangue , Interleucina-6/sangue , Masculino , Ratos Sprague-Dawley , Recuperação de Função Fisiológica , Linfócitos T Reguladores/metabolismo , Células Th17/metabolismo , Fatores de Tempo , Fator de Necrose Tumoral alfa/sangue , Função Ventricular EsquerdaRESUMO
Background: To evaluate the natural innate and adaptive immunity through gene expression and cytology levels in peripheral blood mononuclear cells in patients with acute myocardial infarction (AMI), stable angina pectoris (SAP) and controls. Methods: 210 patients with AMI, 210 with SAP, and 250 clinical controls were recruited. Whole human genome microarray analysis was performed in 20 randomly chosen subjects per group were examined to detect the expressions of complement markers, natural killer cells, T cells and B cells. The quantity of these cells and related cytokines as well as immunoglobulin levels were measured in all subjects. Results: In AMI group, the mRNA expressions of late complement component, markers of natural killer cells, CD3+, CD8+ T cells and B cells were down-regulated, while those of early complement component and CD4+T cells were up-regulated (p<0.05). In both AMI and SAP patients, the quantity of natural killer cells, CD3+, CD8+ T cells, B cells, IgM and IgG were significantly lower than those of the controls. CD4+ T cells, CH50, C3, C4, IL-2, IL-4, IL-6 and IFN-γ were significantly higher (p<0.05). Conclusions: In AMI patients, both of gene expressions related to complement, natural killer cells, CD3+, CD8+ T cells, B cells and the quantity of these immune cells decreased while cell number reduced in SAP patients. Immune function in both AMI and SAP patients decreased especially in AMI patients with declined gene and protein levels. To improve the immune system is a potential target for medical interventions and prevention in AMI.
Assuntos
Angina Estável/imunologia , Infarto do Miocárdio/imunologia , Imunidade Adaptativa/imunologia , Imunidade Adaptativa/fisiologia , Idoso , Angina Estável/sangue , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Feminino , Humanos , Interferon gama/sangue , Interleucina-2/sangue , Interleucina-4/sangue , Interleucina-6/sangue , Masculino , Pessoa de Meia-Idade , Infarto do Miocárdio/sangueRESUMO
BACKGROUND: This clinical observation study aimed to investigate the relationship between the serum levels of vascular endothelial growth factor (VEGF) and its soluble receptors with the severity and the occurrence of late acute respiratory distress syndrome (ARDS) in early trauma. METHODS: Sixty patients with multiple injuries were divided into three groups according to the Injury Severity Score (ISS) and the serum levels of VEGF, soluble VEGF receptor 1 (sVEGFR1), and sVEGFR2, were measured. Ten healthy people were recruited as controls. The incidence of late ARDS was also monitored, and its relationship to the above measures analyzed. RESULTS: VEGF was not associated with ISS (p > 0.05); sVEGFR1 was positively associated with ISS (r = 0.459, p < 0.0001); however, sVEGFR2 was negatively associated with ISS (r = 0.510, p < 0.0001). The serum VEGF levels between the ARDS group and the non-ARDS group showed no significant difference (p > 0.05). sVEGFR1 in the ARDS group was significantly higher than that in the non-ARDS group (p < 0.0001), and sVEGFR2 in the ARDS group was significantly lower than that in the non-ARDS group (p < 0.0001). CONCLUSION: In conclusion, the increasing of sVEGFR1 and the decreasing of sVEGFR2 in early trauma might be closely related to the occurrence of late ARDS. LEVEL OF EVIDENCE: Prognostic study, level III.
Assuntos
Traumatismo Múltiplo/complicações , Receptores de Fatores de Crescimento do Endotélio Vascular/sangue , Síndrome do Desconforto Respiratório/etiologia , Fator A de Crescimento do Endotélio Vascular/sangue , Adulto , Estudos de Casos e Controles , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Escala de Gravidade do Ferimento , Masculino , Pessoa de Meia-IdadeRESUMO
The current study aimed to identify differentially expressed B cellassociated genes in peripheral blood mononuclear cells and observe the changes in B cell activation at different stages of coronary artery disease. Groups of patients with acute myocardial infarction (AMI) and stable angina (SA), as well as healthy volunteers, were recruited into the study (n=20 per group). Whole human genome microarray analysis was performed to examine the expression of B cellassociated genes among these three groups. The mRNA expression levels of 60 genes associated with B cell activity and regulation were measured using reverse transcriptionquantitative polymerase chain reaction. The mRNA expression of the B cell antigen receptor (BCR)associated genes, CD45, NFAM, SYK and LYN, were significantly upregulated in patients with AMI; however, FCRL3, CD79B, CD19, CD81, FYN, BLK, CD22 and CD5 mRNA expression levels were significantly downregulated, compared with patients in the SA and control group. The mRNA levels of the Tindependent B cellassociated genes, CD16, CD32, LILRA1 and TLR9, were significantly increased in AMI patients compared with SA and control patients. The mRNA expression of genes associated with Tdependent B cells were also measured: EMR2 and CD97 were statistically upregulated, whereas SLAMF1, LY9, CD28, CD43, CD72, ICOSL, PD1, CD40 and CD20 mRNAs were significantly downregulated in AMI group patients compared with the two other groups. Additionally the gene expression levels of B cell regulatory genes were measured. In patients with AMI, CR1, LILRB2, LILRB3 and VAV1 mRNA expression levels were statistically increased, whereas, CS1 and IL4I1 mRNAs were significantly reduced compared with the SA and control groups. There was no statistically significant difference in B cellassociated gene expression levels between patients with SA and the control group. The present study identified the downregulation of genes associated with BCRs, B2 cells and B cell regulators in patients with AMI, indicating a weakened T cellB cell interaction and reduced B2 cell activation during AMI. Thus, improving B2 cellmediated humoral immunity may be a potential target for medical intervention in patients with AMI.
Assuntos
Subpopulações de Linfócitos B/imunologia , Doença da Artéria Coronariana/imunologia , Regulação da Expressão Gênica/imunologia , Adulto , Idoso , Subpopulações de Linfócitos B/patologia , Doença da Artéria Coronariana/patologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Infarto do Miocárdio/imunologia , Infarto do Miocárdio/patologiaRESUMO
OBJECTIVE: To investigate the differential gene expression of cytokines and compare their impacts on the immune functions among the acute myocardial infarction patients (AMI), the stable angina patients (SA) and the controls. METHODS: 20 patients with AMI, 20 patients with SA and 20 healthy volunteers were recruited into the study. Whole human genome microarray analysis was used to detect the gene expression differences in interferons, interleukins, chemokines, tumor necrosis factors and associated receptors in peripheral blood mononuclear cells (PBMCs) among three groups. RESULTS: Compared with SA patients and the controls respectively, in AMI patients, IFNα2, IFNαR1, IFNαR2, IFNγR1, IFNγR2, L1ß, IL16, IL18, Cxcl1, Cxcl2, Cxcl6, CxcR2, CxcR4, LIGHT, TNFR1, LT-ßR, CD137, TRAILR, and TWEAKR mRNA expressions were significantly up-regulated (P<0.05), while Ccl5, Ccl24, Ccl28, CcR5, TWEAK, CD40, CD27, and BAFFR mRNA expressions were significantly down-regulated (P<0.05). But, there was no significant difference in cytokine expression between the SA patients and the controls. CONCLUSION: In AMI patients, mRNA expression levels of cytokines were imbalanced, indicating the dysfunction of the immune system. Together with no significant change of cytokines was observed between the SA and controls, showing the different cytokine related immune activity in the AMI and SA patients.
RESUMO
OBJECTIVE: To investigate the effects of exercise therapy at the intensity of anaerobic threshold (AT) for exercise tolerance in patients with chronic stable coronary artery disease. METHODS: Forty-three patients with chronic stable coronary artery disease (3 patients after coronary arterial bypass graft (CABG) surgery, 22 patients with old myocardial infarction and 18 unstable angina pectoris undergoing successful percutaneous coronary intervention (PCI) finished twice cardiopulmonary exercise test (CPET) and followed their rehabilitation program for 3 months. Thirty-two patients finished their aerobic exercise therapy based on their individual anaerobic thresholds while 11 patients had no exercise therapy. RESULTS: The heart rate at AT intensity (97 ± 9/min) was lower than their traditional minimal target heart rate (112 ± 7/min) and lower than heart rate (115 ± 11/min) at ischemic threshold post-CPET. The O(2) consumption (10.7 ± 2.4 to 12.6 ± 2.9 ml×min(-1)×kg(-1)) (P = 0.04) and workload (37 ± 18 to 47 ± 13 J/s) (P = 0.04) at AT level and the O(2) consumption (15.3 ± 3.1 to 20.6 ± 4.2 ml×min(-1)×kg(-1), P = 0.02) and workload(68 ± 12 and 87 ± 14 J/s, P = 0.01) at peak level markedly increased after 3 months in the exercise group. And the O(2) consumption (15.3 ± 2.9 to 16.2 ± 3.1 ml×min(-1)×kg(-1)) and workload (65 ± 13 to 73 ± 16 J/s) at peak level mild increased after 3 months in the non-exercise group, but their O(2) consumption (11.0 ± 2.7 to 11.3 ± 2.8 ml×min(-1)×kg(-1)) and workload (38 ± 11 to 37 ± 9 J/s) at AT level had no obvious change. CONCLUSION: AT exercise intensity was lower than ischemic threshold post-CPET. Exercise therapy at the intensity of anaerobic threshold can improve oxygen capacity and exercise tolerance.
Assuntos
Limiar Anaeróbio , Doença da Artéria Coronariana/metabolismo , Doença da Artéria Coronariana/reabilitação , Terapia por Exercício , Oxigênio/metabolismo , Idoso , Teste de Esforço , Tolerância ao Exercício , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
Cordycepin (3'-deoxyadenosine) is the main functional component of Cordycepins militaris, a renowned traditional Chinese medicine, which has been shown to possess anti-tumor, anti-inflammatory, anti-diabetic and neuro-protective effects. However, the effect of cordycepin on the central nervous system (CNS) remains unclear. In this study, the effects of cordycepin on neuronal activity were investigated on the CA1 pyramidal neurons in rat hippocampal brain slices using a whole-cell patch clamp technique. Our results revealed that cordycepin significantly decreased the frequency of both the spontaneous and evoked action potential (AP) firing. While AP spike width, the amplitude of fast after hyperpolarization (fAHP), and membrane input resistance were not altered by cordycepin, the neuronal membrane potential was hyperpolarized by cordycepin. Collectively, these results demonstrate that cordycepin reduces neuronal activity by inducing membrane hyperpolarization, indicating that cordycepin may be a potential therapeutic strategy for ischemic and other excitotoxic disorders.