Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 11 de 11
Filtrar
Mais filtros










Base de dados
Intervalo de ano de publicação
1.
Acta Histochem ; 121(2): 253-259, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30611528

RESUMO

Understanding the mechanisms of adipogenic differentiation may lead to the discovery of novel therapeutic targets for obesity. The natural plant polyphenol compound curcumin can improve obesity-associated inflammation and diabetes in obese mice. The role of curcumin in adipogenic differentiation of human bone marrow mesenchymal stem cells (hMSCs) is still unclear. We used hMSCs to investigate the details of the mechanism underlying the adipogenic effects of curcumin. At different time points (i.e., 5 days and 10 days) of hMSC adipocyte differentiation, an accumulation of large lipid droplets was analyzed in Oil Red O-stained cultured cells in two curcumin (5 µM and 10 µM) groups and the control group. The cells were also harvested for the detection of mRNA and protein expressions by quantitative real-time polymerase chain reaction and Western blot analysis. The results showed that curcumin can suppresses adipocyte differentiation in a dose-dependent manner and inhibited the expression of PPARγ, C/EBPα, and FABP4. Importantly, curcumin can also suppress the expression of Kruppel-like factor 15, which may bind to the PPARγ promoter, resulting in downregulation of PPARγ expression to inhibit the adipogenic differentiation of hMSCs.


Assuntos
Adipócitos/citologia , Adipogenia/fisiologia , Células da Medula Óssea/citologia , Diferenciação Celular/fisiologia , Células-Tronco Mesenquimais/citologia , Regulação para Baixo/fisiologia , Humanos , Fatores de Transcrição Kruppel-Like/metabolismo , Fatores de Transcrição/metabolismo
2.
Mol Cell Biochem ; 449(1-2): 295-303, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-29959592

RESUMO

MicroRNAs are members of the family of non-coding small RNAs that regulate gene expression either by inhibiting mRNA translation or by promoting mRNA degradation at the post-transcriptional level. They play an important role in the differentiation of human bone marrow mesenchymal stem cells (hMSCs) into adipocytes. However, the role of microRNAs in this process remains to be poorly understood. Here, we observed that miR-377-3p expression was markedly decreased during adipogenic differentiation of hMSCs. Overexpression of miR-377-3p decreased adipocyte differentiation and downregulated the expression of adipogenic markers. Meanwhile, bioinformatics-based studies suggested that LIFR is a target of miR-377-3p. Further analysis confirmed that expression of LIFR present markedly increased during adipogenic differentiation of hMSCs. In addition, downregulation expression of LIFR significantly inhibited the process of adipocyte differentiation. To confirm the relation between miR-377-3p and LIFR, luciferase reporter assays were carried out. The results indicated that miR-377-3p bound directly to the 3'-untranslated region of LIFR. These data indicate that miR-377-3p suppressed adipogenesis of hMSCs by targeting LIFR, which provides novel insights into the molecular mechanism of miRNA-mediated cellular differentiation.


Assuntos
Adipogenia , Células da Medula Óssea/metabolismo , Diferenciação Celular , Subunidade alfa de Receptor de Fator Inibidor de Leucemia/biossíntese , Células-Tronco Mesenquimais/metabolismo , MicroRNAs/metabolismo , Regiões 3' não Traduzidas , Células da Medula Óssea/citologia , Linhagem Celular , Humanos , Subunidade alfa de Receptor de Fator Inibidor de Leucemia/genética , Células-Tronco Mesenquimais/citologia , MicroRNAs/genética
3.
Mol Med Rep ; 15(4): 1571-1576, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28260060

RESUMO

The present study aimed to screen several differentially expressed genes (DEGs) and differentially expressed microRNAs (miRNAs) for two types of mesenchymal stem cell (MSC) differentiation. Bone morphogenetic protein 6 (BMP­6) and dexamethasone were used to induce MSCs towards osteoblastic differentiation or adipocytic differentiation. The t­test in the Bioconductor bioinformatics software tool was used to screen DEGs and differentially expressed miRNAs in the two samples. Subsequent gene ontology (GO) and pathway analyses on the DEGs were performed using the GO and Kyoto Encyclopedia of Genes and Genomes databases, respectively; potential target genes for the screened miRNAs were predicted using the TargetScan database. In addition, an interaction network between the DEGs and miRNAs was constructed. Numerous DEGs and miRNAs were screened during osteoblastic and adipocytic differentiation of MSCs. Important pathways, such as glutathione metabolism, pathogenic Escherichia coli infection and Parkinson's disease, and GO terms, including cytoskeletal protein binding and phospholipase inhibitor activity, were enriched in the screened DEGs from MSCs undergoing osteogenic differentiation and adipocytic differentiation. miRNAs, including miRNA (miR)­382 and miR­203, and DEGs, including neuronal growth regulator 1 (NEGR1), phosphatidic acid phosphatase 2B (PPAP2B), platelet­derived growth factor receptor alpha (PDGFRA), interleukin 6 signal transducer (IL6ST) and sortilin 1 (SORT1), were demonstrated to be involved in osteoblastic differentiation. In addition, the downregulated miRNAs (including miR­495, miR­376a and miR­543), the upregulated miR­106a, the upregulated DEGs, including enabled homolog (ENAH), polypeptide N­acetylgalactosaminyltransferase 1 and acyl­CoA synthetase long­chain family member 1, and the downregulated repulsive guidance molecule family member B and semaphorin SEMA7A were demonstrated to be involved in adipocytic differentiation. The results of the present study suggested that miRNAs (miR­203 and miR­382) and DEGs (NEGR1, PPAP2B, PDGFRA, IL6ST and SORT1) may serve pivotal functions in the osteoblastic differentiation of MSCs, whereas miR­495, which is also involved in osteoblast differentiation and had four targets, including NEGR1, miR­376a, miR­543 and ENAH may have crucial roles in adipocytic differentiation of MSCs.


Assuntos
Adipócitos/citologia , Diferenciação Celular , Biologia Computacional/métodos , Células-Tronco Mesenquimais/citologia , Osteoblastos/citologia , Adipócitos/metabolismo , Diferenciação Celular/genética , Perfilação da Expressão Gênica , Ontologia Genética , Redes Reguladoras de Genes , Humanos , Células-Tronco Mesenquimais/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Osteoblastos/metabolismo , Mapas de Interação de Proteínas/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Transdução de Sinais/genética
4.
Folia Histochem Cytobiol ; 54(1): 14-24, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27044590

RESUMO

INTRODUCTION: Adipogenesis comprises multiple processes by which mesenchymal stem cells differentiate into adipocytes. To increase our knowledge of the mechanism underlying adipogenic differentiation of human bone marrow mesenchymal stem cells (hMSCs), we performed full-genome gene expression microarray and gene ontology analyses of induced differentiation of hMSCs. MATERIAL AND METHODS: Adipogenic differentiation of hMSCs was induced by an adipogenic medium, and total RNA was extracted from undifferentiated hMSCs (day 0) and differentiated adipocytes (day 14). Then microarray hybridization of RNA samples was performed. The GeneChip Operating Software was used to analyze the hybridization data to identify differentially expressed genes, which were performed Gene Ontology categorization and pathway analysis. Pathway-act-network and genes-act-network were built according to the Kyoto Encyclopedia of Genes and Genomes database. Some differentially expressed genes were subjected to qRT-PCR to verify the microarray data. RESULTS: We detected a total of 3,821 differentially expressed genes, of which 753 were upregulated and 3,068 downregulated. These genes were well represented in a variety of functional categories, including collagen fibril organization, brown fat cell differentiation, cell division, and S phase of mitotic cell cycle. Subsequently, pathway analysis was conducted, and significant pathways (from top 50) were selected for pathway-act-network analysis, which indicated that the mitogen-activated protein kinase (MAPK) pathway and cell cycle were of high degrees (> 10). Gene-act-network analysis showed that insulin-like growth factor 1 receptor (IGF1R), histone deacetylase 1 (HDAC1), HDAC2, MAPK13, MAPK8, phosphoinositide-3-kinase regulatory subunit 1 (PI3KR1), and PI3KR2 also had high degrees (> 18). CONCLUSIONS: Collectively, these data provide novel information and could serve as a basis for future study to clarify the mechanisms underlying adipocyte differentiation of hMSCs.


Assuntos
Adipogenia/genética , Células da Medula Óssea/fisiologia , Células-Tronco Mesenquimais/fisiologia , Adipócitos/citologia , Adipócitos/metabolismo , Adipócitos/fisiologia , Adipócitos Marrons/citologia , Adipócitos Marrons/metabolismo , Adipócitos Marrons/fisiologia , Células da Medula Óssea/citologia , Células da Medula Óssea/metabolismo , Diferenciação Celular , Células Cultivadas , Perfilação da Expressão Gênica , Humanos , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , RNA/genética , RNA/metabolismo , Reação em Cadeia da Polimerase em Tempo Real
5.
Nan Fang Yi Ke Da Xue Xue Bao ; 37(2): 199-203, 2016 Feb 20.
Artigo em Chinês | MEDLINE | ID: mdl-28219863

RESUMO

OBJECTIVE: To screen the differentially expressed miRNAs and their target genes in adipogenic differentiation of human bone marrow mesenchymal stem cells (hMSCs) to better understand the mechanism for regulating the balance between osteoblast and adipocyte differentiation. METHODS: Cultured hMSCs were induced for adipogenic differentiation, and at 0, 7, 14, and 21 days of induction, the cells were examined for miRNA and mRNA expression profiles using miRNA chip and transcriptome sequencing (RNA-seq) techniques. Correlation analysis was carried out for the miRNAs and mRNAs of potential interest. The databases including TargetScan, PicTar and miRanda were used to predict the target genes of the differentially expressed miRNA. RESULTS: The expression of miR-140-5p was down-regulated and leukemia inhibitory factor receptor (LIFR) expression increased progressively during adipogenic differentiation of hMSCs, showing a negative correlation between them. Target gene prediction using the 3 databases identified LIFR as the target gene of miR-140-5p. CONCLUSION: miRNA-140-5p may play an important role by regulating its target gene LIFR during adipogenic differentiation of hMSCs.


Assuntos
Adipogenia , Diferenciação Celular , Células-Tronco Mesenquimais/citologia , MicroRNAs/genética , Adipócitos/citologia , Células Cultivadas , Regulação para Baixo , Humanos , Subunidade alfa de Receptor de Fator Inibidor de Leucemia/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos , Osteoblastos/citologia , RNA Mensageiro , Transcriptoma
6.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 30(7): 717-20, 2014 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-25001936

RESUMO

OBJECTIVE: To verify whether insulin promotes the differentiation of skeletal myoblasts into myocytes and whether wortmannin and U0126 inhibit the promoting effect with an attempt to explore the molecular mechanisms of inducing the differentiation of muscular cells in rats. METHODS: Primary skeletal myoblasts were separated and cultured from rats, and then were treated in DMEM containing various concentrations of insulin. The morphology of cells was monitored under a phase-contrast microscope. And the expression of myogenin was detected by immunocytochemistry and Western blotting. The change in the effect of insulin on the differentiation of myoblasts was observed after the intervention of a phosphatidylinositide 3-kinase (PI3K) inhibitor wortmannin and a specific MEK inhibitor U0126. RESULTS: Insulin markedly promoted myotube formation of myoblasts. Two days after insulin treatment, myotubes started to form; later, more and more myotubes appeared, and to the peak at 7 days. Insulin increased the expression of myogenin in a concentration-dependent manner. However, wortmannin and U0126 inhibited the effect of insulin on the differentiation of skeletal myoblasts in rats. CONCLUSION: Wortmannin and U0126 can suppress the promoting effect of insulin on the differentiation of skeletal myoblasts into myocytes in rats and decrease the formation of myotubes and the expression of myogenin.


Assuntos
Androstadienos/farmacologia , Butadienos/farmacologia , Diferenciação Celular/efeitos dos fármacos , Insulina/farmacologia , Mioblastos Esqueléticos/efeitos dos fármacos , Nitrilos/farmacologia , Animais , Western Blotting , Células Cultivadas , Relação Dose-Resposta a Droga , Imuno-Histoquímica , MAP Quinase Quinase 1/antagonistas & inibidores , MAP Quinase Quinase 1/metabolismo , Microscopia de Contraste de Fase , Fibras Musculares Esqueléticas/citologia , Fibras Musculares Esqueléticas/efeitos dos fármacos , Fibras Musculares Esqueléticas/metabolismo , Mioblastos Esqueléticos/citologia , Mioblastos Esqueléticos/metabolismo , Miogenina/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Ratos , Wortmanina
7.
Exp Parasitol ; 135(3): 497-502, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-23999146

RESUMO

Schizophrenia is a serious neuropsychiatric disease of uncertain etiology, which causes human mental disorder and affects about 1% of the population. In recently years, some studies showed that some cases of schizophrenia may be associated with Toxoplasma gondii infection. In order to investigate a potential association between Toxoplasma infection and schizophrenia, we investigated the relative clinical symptom of schizophrenia such as learning and memory capability, depression and stereotypy to find some useful information by behavioral test in mouse models. Our results demonstrated that mice from Toxoplasma infection and MK-801 administration (as the model of schizophrenia) were impaired in learning and memory capability, and they had more serious depression and stereotypy compared with the control mice, especially the mice from congenital Toxoplasma infection. In addition, our results clearly showed that the number of cysts in brain tissue of congenital Toxoplasma infection mice was significantly low than in acquired Toxoplasma infected mice. Collectively, these results suggested a potential association between Toxoplasma infection and schizophrenia.


Assuntos
Esquizofrenia/parasitologia , Toxoplasmose Cerebral/complicações , Toxoplasmose Congênita/complicações , Animais , Aprendizagem da Esquiva , Comportamento Animal , Encéfalo/parasitologia , Modelos Animais de Doenças , Maleato de Dizocilpina , Antagonistas de Aminoácidos Excitatórios , Feminino , Aprendizagem , Masculino , Memória , Camundongos , Camundongos Endogâmicos BALB C , Esquizofrenia/induzido quimicamente , Comportamento Estereotipado , Toxoplasmose Congênita/parasitologia
8.
Zhong Yao Cai ; 35(9): 1382-5, 2012 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-23451489

RESUMO

OBJECTIVE: Using papain to prepare polypeptide from Eupolyphaga sinensis, then study the immune function of polypeptide from Eupolyphaga sinensis in vivo. METHODS: Used hydrolysis degree as index, pH value, enzyme dosage, thermometer reaction time were optimized. Studied the influence of polyeptide on the mice immune functions through mice immune organs index, phagocytic function and the level of IL-2. RESULTS: The optimum enzymolysis condition was as follows: pH 8.0, enzyme 1%, temperature 55 degrees C, reaction time 4. 5 h. In vivo test of mice demonstraed that, Eupolyphaga sinensis could elevate index of thymus and spleen, enhance the phagocytic function of macrophage and promote the level of IL-2 in serum. CONCLUSION: Eupolyphaga sinensis has immunoregulatory effect.


Assuntos
Baratas/química , Papaína/metabolismo , Peptídeos/imunologia , Peptídeos/isolamento & purificação , Tecnologia Farmacêutica/métodos , Animais , Feminino , Concentração de Íons de Hidrogênio , Hidrólise , Interleucina-2/sangue , Camundongos , Papaína/química , Peptídeos/farmacologia , Baço/efeitos dos fármacos , Baço/imunologia , Temperatura Ambiente , Timo/efeitos dos fármacos , Timo/imunologia
9.
Sheng Li Xue Bao ; 63(4): 300-4, 2011 Aug 25.
Artigo em Chinês | MEDLINE | ID: mdl-21861047

RESUMO

The aim of this study was to investigate the influence of neonatal isolation stress on hyperlocomotion in complexin II knockout mouse (Cplx2(-/-)). The mice were randomly divided into 4 groups: Cplx2(-/-) with stress, Cplx2(+/+) with stress, Cplx2(-/-) without stress and Cplx2(+/+) without stress. Isolation stress was employed on the pups of stress groups from the 2nd day after the postnatal to the 21st day. The PCR was used to determine the gene type and the hyperlocomotion test was employed to detect the change of animal behavior after methamphetamine or saline injection (i.p.). The results showed that the animals of all groups increased their movement after injection of 0.2 mg/kg methamphetamine in different levels (P < 0.01), compared with those injected with saline. The Cplx2(-/-) mouse with stress revealed a significant increase in the distance of free movement after injection of 0.2 mg/kg methamphetamine compared with the knockout mouse without stress (P < 0.001). When Cplx2(-/-) mouse with stress was compared with wild type with stress, Cplx2(-/-) mouse with stress had more movement (P < 0.001), indicating that Cplx2 has effect on the hyperlocomotion as well. These results suggest an involvement of stress and Cplx2 in the movement behavior of mice.


Assuntos
Proteínas Adaptadoras de Transporte Vesicular/genética , Locomoção/fisiologia , Proteínas do Tecido Nervoso/genética , Isolamento Social , Estresse Psicológico/psicologia , Animais , Animais Recém-Nascidos , Comportamento Animal/fisiologia , Metanfetamina/farmacologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos Mutantes
10.
Zhongguo Zhong Yao Za Zhi ; 35(15): 1925-30, 2010 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-20931838

RESUMO

OBJECTIVE: To obtain the cDNA sequence encoding fibrinolytic enzyme from Eupolyphaga sinensis and express it in prokaryotic and eukaryotic expression system. METHOD: The primers were designed according to the cDNA of other animals'fibrinolytic enzyme. The cDNA sequence was cloned by RT-PCR and 3 RACE. RESULT: Sequence analysis revealed that the length of the cDNA fragment was 672 bp and encoded a protein of 224 amino acid residues, the N end amino acid sequence residues was IVGG in accordance with other fibrinolytic enzyme. The cDNA sequence was expressed in E. coli, inactive protein was obtained. While expressed in Pichia pastoris, recombinant protein had fibrinolytic activity. CONCLUSION: The cDNA sequence of fibrinolytic enzyme from E. sinensis Walker was cloned and expressed for the first time and it proved a good basis for further functional study of the enzyme.


Assuntos
Clonagem Molecular , Baratas/enzimologia , Fibrinolisina/genética , Expressão Gênica , Proteínas de Insetos/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Baratas/química , Baratas/genética , DNA Complementar/química , DNA Complementar/genética , Fibrinolisina/química , Fibrinolisina/metabolismo , Proteínas de Insetos/química , Proteínas de Insetos/metabolismo , Dados de Sequência Molecular , Alinhamento de Sequência
11.
Zhong Yao Cai ; 32(6): 846-8, 2009 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-19764321

RESUMO

OBJECTIVE: To obtain polysaccharide from Scolopendra subspinipes mutilans and study its partial properties. METHODS: A raw polysaccharide was isolated from Chinese medicine Scolopendra subspinipes mutilans, and three groups were isolated by DEAE-52. The thin layer chromatography and gel filtration chromatography were used to detect the main monosaccharide composition and molecular weight of the group I. The inhibitory effect of the group I on tumor cells was detected by MTT assay. RESULTS: The molecular weight of the group I was 33.1 kD. The inhibitory effect of the group I on Hela cells was obvious, as its inhibitive rate was 60.8% on Hela cells when the polysaccharide's concentration was 3.13 microg/mL, but it had no effect on the Eca 109 cells. CONCLUSION: The group I has specific effects on different tumor cells.


Assuntos
Antineoplásicos/farmacologia , Artrópodes/química , Polissacarídeos/isolamento & purificação , Polissacarídeos/farmacologia , Animais , Proliferação de Células/efeitos dos fármacos , Neoplasias Esofágicas/patologia , Células HeLa , Humanos , Materia Medica/química , Materia Medica/isolamento & purificação , Materia Medica/farmacologia , Peso Molecular , Polissacarídeos/química , Espectrofotometria Ultravioleta
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA