RESUMO
Our study aimed to investigate the effects of interleukin-4 (IL-4) on macrophage polarization, as well as its role in the development of atherosclerosis. Human peripheral blood mononuclear cells (PBMCs) were isolated and randomly divided into 3 groups: control group, ox-LDL group, and ox-LDL + IL-4 groups. The expression of M1/M2 macrophage surface markers such as TNF-α, CD68, and CD206 were analyzed by western blot. Cell viability was determined using the MTT assay. Measurement of CD86/CD206 expression ratio (M1/M2 ratio) was performed via flow cytometry. In addition, ApoE(-/-) mice on a C57BL/6 background were subjected to high-fat diets, and were used as a model of atherosclerosis. Atherosclerotic lesion area was quantified after mice were treated with ox-LDL and IL-4. Finally, expression of phosphorylated MAPK signaling molecules such as p-ERK and p-JNK was quantified using western blot. The expression of TNF-α and CD86 markedly increased after cells were treated with ox-LDL, whereas the expression of CD206 markedly increased after PBMCs were treated with IL-4. It is possible that IL-4 could decrease ox-LDL-induced cell viability and the CD86/CD206 (M1/M2) ratio. Additionally, IL-4 intervention attenuated ox-LDL-induced atherosclerotic lesions in ApoE(-/-) mice, and decreased ox- LDL-induced expression of p-ERK and p-JNK. Our findings indicate that IL-4 may induce macrophages to take on an M2 phenotype in order to resolve inflammation via inhibition of MAPK signaling pathways, thereby protecting against atherosclerosis. IL-4 may serve as an intervention target for atherosclerosis.
Assuntos
Aterosclerose/tratamento farmacológico , Aterosclerose/metabolismo , Interleucina-4/farmacologia , Interleucina-4/uso terapêutico , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Animais , Apolipoproteínas E/metabolismo , Células Cultivadas , Lipoproteínas LDL/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Fosforilação/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacosRESUMO
We evaluated the effects of down-regulated heme oxygenase (HO)-1 expression on the proliferation of the acute myelocytic leukemia Kasumi-1 cell line by using the HO-1 inhibitor zinc protoporphyrin IX (ZnPPIX) in combination with daunorubicin (DNR), and evaluated the mechanism. The proliferation rates of cells treated with 10 mg/mL DNR and 10 mM ZnPPIX individually or in combination for different time periods were detected using the MTT assay. The apoptotic outcomes of the blank control, ZnPPIX, DNR, and ZnPPIX groups in combination with the DNR group were detected by flow cytometry. The expression of HO-1, activating transcription factor 4, CCAAT-enhancer-binding protein homologous protein, and inositol-requiring enzyme-α mRNA and proteins were detected by fluorescent quantitative real-time polymerase chain reaction and western blotting, respectively. Combined administration inhibited the cells most potently and time-dependently, decreased the expression of HO-1, and significantly increased the expression of activating transcription factor 4, CCAAT-enhancer-binding protein homologous protein, and inositol-requiring enzyme-α expression levels. The cell apoptotic rates in the blank control, DNR, ZnPPIX, and combined administration groups were 8.32 ± 0.53, 39.16 ± 1.46, 10.46 ± 0.88, and 56.26 ± 2.24%, respectively. Inhibiting HO-1 expression can enhance the damaging effects of DNR on Kasumi-1 cells, providing experimental evidence for the improvement of therapeutic effects on acute myelocytic leukemia in clinical practice.
Assuntos
Fator 4 Ativador da Transcrição/biossíntese , Endorribonucleases/biossíntese , Heme Oxigenase-1/biossíntese , Leucemia Mieloide Aguda/genética , Proteínas Serina-Treonina Quinases/biossíntese , Fator de Transcrição CHOP/biossíntese , Fator 4 Ativador da Transcrição/genética , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Endorribonucleases/genética , Inibidores Enzimáticos/administração & dosagem , Regulação Leucêmica da Expressão Gênica/efeitos dos fármacos , Heme Oxigenase-1/antagonistas & inibidores , Heme Oxigenase-1/genética , Humanos , Leucemia Mieloide Aguda/tratamento farmacológico , Leucemia Mieloide Aguda/patologia , Proteínas Serina-Treonina Quinases/genética , RNA Mensageiro/biossíntese , Transdução de Sinais/efeitos dos fármacos , Fator de Transcrição CHOP/genéticaRESUMO
We sought to investigate the effect of nerve growth factor (NGF) expression on the formation and prognosis of cerebral aneurysms. Forty-eight cases were selected following a diagnosis of cerebral aneurysm using computed tomography angiography and surgical confirmation. Thirty-four cases of healthy deaths were also chosen. The tissue was tested for NGF expression changes by reverse-transcription PCR, Western blot and histopathology, and NGF expression was compared between the cerebral aneurysm and healthy groups. The expression level of NGF in cerebral aneurysm tissue was significantly increased over that observed in control tissue. The abnormal expression of NGF is related to cerebral aneurysms. The elevated expression of NGF in cerebral aneurysms may be associated with a poor prognosis.
Assuntos
Angiografia Cerebral/métodos , Aneurisma Intracraniano/diagnóstico por imagem , Aneurisma Intracraniano/patologia , Fator de Crescimento Neural/metabolismo , Idoso , Feminino , Humanos , Aneurisma Intracraniano/mortalidade , Masculino , Pessoa de Meia-Idade , Fator de Crescimento Neural/biossíntese , Prognóstico , Tomografia Computadorizada por Raios XRESUMO
We examined the function of survivin gene expression in patients with nasopharyngeal carcinoma (NPC), as well as small interfering RNA (siRNA) on controlling CNE-2 NPC proliferation and apoptosis. Immunohistological methods, in situ hybridization, and reverse transcription-polymerase chain reaction technique were used to detect survivin protein and mRNA expression. We designed an siRNA sequence to inhibit survivin gene expression. The MTT method was used to examine the function of siRNA on controlling cell growth and proliferation. Induction of cell apoptosis by siRNA was examined by flow cytometry; electron microscopy was used to observe ultrastructure changes in CNE-2 cells. Western blotting was used to detect survivin gene expression. The survivin protein was expressed in 71.9% of cells, while its mRNA was expressed in 65.6% of cells. Relative mRNA expression was 4.16 x 10(-2); these data for the control groups were 23.3, 33.3, and 4.42 x 10(-4), respectively. Following transfection with 3 different siRNA sequences, survivin mRNA expression in CNE-2 cells was decreased. Inhibition of cell proliferation and rate of apoptosis increased with increasing siRNA concentration. Western blotting revealed decreased survivin expression and electron microscopy revealed ultrastructural changes in cancer cells. Survivin gene expression in NPC generally increased. In vitro transcription of siRNA decreased CNE-2 survivin gene expression, and different sequences of siRNA decrease gene expression in CNE-2 cells to varying degrees. Transfected siRNA3 can effectively inhibit CNE-2 cell proliferation and induce apoptosis; gene silencing using siRNA may represent a new treatment for NPC.
Assuntos
Apoptose/genética , Proteínas Inibidoras de Apoptose/genética , Neoplasias Nasofaríngeas/genética , Interferência de RNA , RNA Interferente Pequeno , Adulto , Idoso , Carcinoma , Proliferação de Células/genética , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Carcinoma Nasofaríngeo , Neoplasias Nasofaríngeas/metabolismo , Survivina , Adulto JovemRESUMO
OBJECTIVES: The present study is to evaluate the expression level of enhancer of zeste homolog 2 (EZH2) and vascular endothelial growth factor (VEGF), and analyze their correlations with clinicopathological characteristics and survival in patients with clear cell renal cell carcinoma (CCRCC). The effect of EZH2 on apoptosis and cell proliferation in 786-O renal cancer cell line is investigated. METHODS: The expression level of EZH2 and VEGF was detected in 185 primary CCRCC patients' tissues using tissue microarray and immunohistochemistry. Small interfering RNA or enhanced green fluorescent protein transfection was employed to investigate the effect of EZH2 inhibition or overexpression on VEGF expression, apoptosis and cell proliferation in 786-O cells using flow cytometry, immunofluorescence microscopy, quantitative real-time reverse-transcription polymerase chain reaction and Western blot analysis. RESULTS: High expression level of EZH2 and VEGF was observed in advanced CCRCC and correlated with the TNM stage (p = 0.013, p = 0.001) and distant metastasis (p = 0.011, p = 0.038), respectively. EZH2 was positively correlated with VEGF in CCRCC tissues (correlation coefficient = 0.850, p < 0.001). Kaplan-Meier survival analysis revealed that patients with positive EZH2 expression had a shorter overall survival time compared to patients with negative EZH2 expression (34.3 vs. 67.2, p < 0.001). In 786-O cells, EZH2 silencing inhibited VEGF expression and cell proliferation while increasing apoptosis (p < 0.001). EZH2 overexpression promoted VEGF expression and cell proliferation while inhibiting apoptosis (p < 0.001). CONCLUSIONS: EZH2 correlates positively with VEGF and associates with adverse clinicopathologic characteristics and shorter survival time in CCRCC patients. EZH2 accelerates antiapoptosis and cell cycle in 786-O cells.
Assuntos
Carcinoma de Células Renais/metabolismo , Regulação Neoplásica da Expressão Gênica , Neoplasias Renais/metabolismo , Complexo Repressor Polycomb 2/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Idoso , Apoptose , Ciclo Celular , Linhagem Celular Tumoral , Proliferação de Células , Progressão da Doença , Proteína Potenciadora do Homólogo 2 de Zeste , Feminino , Citometria de Fluxo , Inativação Gênica , Proteínas de Fluorescência Verde/química , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , RNA Interferente Pequeno/metabolismo , Análise Serial de TecidosRESUMO
This study aimed to investigate the influence of hypercholesterolemia (HC) on intracellular calcium ion concentration in the sphincter of Oddi (SO) of rabbits and the influence of paeoniflorin on intracellular calcium ion concentration in the hypercholesterolemic rabbit SO. Sixteen purebred New Zealand rabbits were randomly divided into two groups: the control group and the HC model group (8 rabbits in each group). The control group was fed standard diet. The HC group was fed standard diet plus cholesterol for a total of 8 weeks to induce and establish the rabbit HC model. The SO segment of HC rabbits was taken and enzyme treated to obtain SO cells. After primary culture, immunohistochemical analysis was performed. Fluo-3/AM was used to load SO cells, and then intracellular calcium ion concentration was determined by confocal microscopy. Intracellular calcium ion in the SO of the HC group was higher than that of the normal group; intracellular calcium ion in the HC rabbit SO of the paeoniflorin group was lower than that of the control group, where the paeoniflorin effect was greater with higher concentrations. High cholesterol caused an increase in intracellular calcium ion concentration in the rabbit SO, and paeoniflorin can reduce intracellular calcium ion concentration in the HC rabbit SO in a concentration-dependent manner.
Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Cálcio/metabolismo , Células Epiteliais/efeitos dos fármacos , Glucosídeos/farmacologia , Hipercolesterolemia/metabolismo , Monoterpenos/farmacologia , Esfíncter da Ampola Hepatopancreática/efeitos dos fármacos , Compostos de Anilina , Animais , Colesterol/metabolismo , Relação Dose-Resposta a Droga , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Feminino , Corantes Fluorescentes , Hipercolesterolemia/patologia , Transporte de Íons/efeitos dos fármacos , Masculino , Cultura Primária de Células , Coelhos , Esfíncter da Ampola Hepatopancreática/metabolismo , Esfíncter da Ampola Hepatopancreática/patologia , XantenosRESUMO
Curcumin has been widely used for the prevention and treatment of Alzheimer's disease (AD), but its mechanism is still not clear. Inhibitory factors of axonal regeneration have been shown to cause a series of pathophysiological changes in the early period of AD. In this study, the co-receptor (Nogo receptor; NgR) of three axonal growth-inhibitory proteins was examined, and effects of curcumin on spatial learning and memory abilities and hippocampal axonal growth were investigated in amyloid ß-protein (Aß)1-40-induced AD rats. Results showed that the expression of NgR in the AD group significantly increased and the number of axonal protein-positive fibers significantly reduced. The spatial learning and memory abilities of AD rats were significantly improved in the curcumin group. Furthermore, hippocampal expressions of NgR mRNA and protein decreased, and the expression of axonal protein significantly increased. There was a negative correlation between the expression of NgR and axonal growth. Together, these results suggested that curcumin could improve the spatial learning and memory abilities of AD rats. The mechanism might be related with its lowering of hippocampal NgR expression and promoting axonal regeneration.
Assuntos
Doença de Alzheimer/metabolismo , Axônios/metabolismo , Curcumina/farmacologia , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Proteínas da Mielina/metabolismo , Receptores de Superfície Celular/metabolismo , Doença de Alzheimer/tratamento farmacológico , Doença de Alzheimer/genética , Doença de Alzheimer/fisiopatologia , Peptídeos beta-Amiloides/efeitos adversos , Animais , Axônios/patologia , Curcumina/administração & dosagem , Modelos Animais de Doenças , Proteínas Ligadas por GPI/genética , Proteínas Ligadas por GPI/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Hipocampo/patologia , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Proteínas da Mielina/genética , Receptor Nogo 1 , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Receptores de Superfície Celular/genéticaRESUMO
This study aimed to analyze the impact factors and outcome of antithrombotic therapy in elderly patients over 65 years old that suffered from atrial fibrillation (AF). A total of 256 elderly patients with AF over 65 years old were divided into 3 groups: 65-74 years old (N = 86), 75-84 years old (N = 122), and over 85 years old (N = 48). The clinical characteristics, antithrombotic therapy, and its related impact factors were retrospectively analyzed. Of all patients, 187 received antithrombotic therapy. In the 65-74 year-old group, 78 patients received antiplatelet treatment (90.7%) and 5 patients received anticoagulation treatment (5.8%). In the 75-84 year-old group, 76 patients received antiplatelet treatment (62.3%) and 14 patients received anticoagulation treatment (11.5%). In the group of over 85 year-olds, 33 patients received antiplatelet therapy (68.8%) and 4 patients received anticoagulation treatment (8.3%). Eleven patients had deep vein thrombosis and atrial thrombosis during antiplatelet therapy (5.9%), 5 patients had gastrointestinal hemorrhage after antiplatelet therapy (2.7%), 2 patients had gastrointestinal bleeding, and 3 patients had brain hemorrhage after anticoagulation treatment (21.7%). Suboptimal antithrombotic therapy was observed in the elderly patients with AF, partly owing to the risks of both thromboembolism and bleeding.
Assuntos
Anticoagulantes/administração & dosagem , Fibrilação Atrial/tratamento farmacológico , Fibrinolíticos/administração & dosagem , Varfarina/administração & dosagem , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Fibrilação Atrial/patologia , Feminino , Fibrinolíticos/efeitos adversos , Hemorragia/induzido quimicamente , Hemorragia/patologia , Humanos , Masculino , Acidente Vascular Cerebral/induzido quimicamente , Acidente Vascular Cerebral/patologia , Resultado do Tratamento , Varfarina/efeitos adversosRESUMO
This study aimed at investigating the ability of cartilage-derived morphogenetic protein 1 (CDMP1) gene-transfected bone marrow mesenchymal stem cells (BMSCs) loaded on the poly(lactic-co-glycolic acid) (PLGA) scaffold for the repair of laryngeal cartilage defects and make a preliminary assessment of its repair effect. The mRNA and protein expressions of hCDMP1 were detected by reverse transcriptase-polymerase chain reaction and Western blotting. The expression of type II collagen (Col II) and glycosaminoglycan (GAG) were detected by immunohistochemistry. The cytoskeletal culture systems before and after transfection were transplanted into the rabbit full-thickness defects of thyroid cartilage for observation of the repair of cartilage defects from general and histological aspects. The exogenous hCDMP1 gene could be successfully transplanted into BMSCs through adenovirus infection to obtain a stable expression. Compared with the control group, hCDMP1 gene-transfected BMSCs had enhanced secretory abilities of Col II, GAG, and other cartilage-specific matrices, with a trend of promoting cartilage differentiation. The transfected cytoskeletal complexes could more effectively repair laryngeal cartilage defects. hCDMP1 gene-transfected BMSCs/PLGA 3-D biological scaffold compounds transplanted into animal bodies could effectively repair laryngeal cartilage defects.