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1.
Cell Mol Neurobiol ; 36(7): 1023-34, 2016 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-27008429

RESUMO

Here, we have investigated the synergistic effect of quercetin administration and transplantation of human umbilical cord mesenchymal stromal cells (HUMSCs) following middle cerebral artery occlusion in rat. Combining quercetin treatment with delayed transplantation of HUMSCs after local cerebral ischemia significantly (i) improved neurological functional recovery; (ii) reduced proinflammatory cytokines (interleukin(IL)-1ß and IL-6), increased anti-inflammatory cytokines (IL-4, IL-10, and transforming growth factor-ß1), and reduced ED-1 positive areas; (iii) inhibited cell apoptosis (caspase-3 expression); and (iv) improved the survival rate of HUMSCs in the injury site. Altogether, our results demonstrate that combined HUMSC transplantation and quercetin treatment is a potential strategy for reducing secondary damage and promoting functional recovery following cerebral ischemia.


Assuntos
Isquemia Encefálica/terapia , Citocinas/metabolismo , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/citologia , Quercetina/farmacologia , Cordão Umbilical/citologia , Animais , Isquemia Encefálica/metabolismo , Modelos Animais de Doenças , Feminino , Humanos , Interleucina-10/metabolismo , Interleucina-4/metabolismo , Interleucina-6/metabolismo , Transplante de Células-Tronco Mesenquimais/métodos , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/imunologia , Ratos Sprague-Dawley
2.
Cell Mol Neurobiol ; 33(4): 465-75, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-23478940

RESUMO

Human mesenchymal stem cells (MSCs) are considered a promising tool for cell-based therapies of nervous system diseases. Bone marrow (BM) has been the traditional source of MSCs (BM-MSCs). However, there are some limitations for their clinical use, such as the decline in cell number and differentiation potential with age. Recently, amniotic fluid (AF)-derived MSCs (AF-MSCs) have been shown to express embryonic and adult stem cell markers, and can differentiate into cells of all three germ layers. In this study, we isolated AF-MSCs from second-trimester AF by limiting dilution and compared their proliferative capacity, multipotency, neural differentiation ability, and secretion of neurotrophins to those of BM-MSCs. AF-MSCs showed a higher proliferative capacity and more rapidly formed and expanded neurospheres compared to those of BM-MSCs. Both immunocytochemical and quantitative real-time PCR analyses demonstrated that AF-MSCs showed higher expression of neural stemness markers than those of BM-MSCs following neural stem cell (NSC) differentiation. Furthermore, the levels of brain-derived growth factor and nerve growth factor secreted by AF-MSCs in the culture medium were higher than those of BM-MSCs. In addition, AF-MSCs maintained a normal karyotype in long-term cultures after NSC differentiation and were not tumorigenic in vivo. Our findings suggest that AF-MSCs are a promising and safe alternative to BM-MSCs for therapy of nervous system diseases.


Assuntos
Líquido Amniótico/citologia , Células da Medula Óssea/citologia , Células-Tronco Mesenquimais/citologia , Neurogênese , Neurônios/citologia , Adulto , Animais , Biomarcadores/metabolismo , Células da Medula Óssea/metabolismo , Proliferação de Células , Separação Celular , Forma Celular , Transformação Celular Neoplásica/patologia , Instabilidade Cromossômica , Cromossomos de Mamíferos/metabolismo , Humanos , Imunofenotipagem , Cariotipagem , Células-Tronco Mesenquimais/metabolismo , Camundongos , Pessoa de Meia-Idade , Células-Tronco Multipotentes/citologia , Células-Tronco Multipotentes/metabolismo , Fatores de Crescimento Neural/metabolismo , Células-Tronco Neurais/citologia , Células-Tronco Neurais/metabolismo , Adulto Jovem
3.
Neurosci Lett ; 525(2): 129-34, 2012 Sep 13.
Artigo em Inglês | MEDLINE | ID: mdl-22902990

RESUMO

Tenascin-R (TN-R) is a neural specific protein and an important molecule involved in inhibition of axonal regeneration after spinal cord injury (SCI). Here we report on rabbit-derived TN-R polyclonal antibody, which acts as a TN-R antagonist with high titer and high specificity, promoted neurite outgrowth and sprouting of rat cortical neurons cultured on the inhibitory TN-R substrate in vitro. When locally administered into the lesion sites of rats received spinal cord dorsal hemisection, these TN-R antibodies could significantly decrease RhoA activation and improve functional recovery from corticospinal tract (CST) transection. Thus, passive immunotherapy with specific TN-R antagonist may represent a promising repair strategy following acute SCI.


Assuntos
Anticorpos/farmacologia , Axônios/efeitos dos fármacos , Traumatismos da Medula Espinal/terapia , Tenascina/antagonistas & inibidores , Animais , Animais Recém-Nascidos , Anticorpos/uso terapêutico , Axônios/fisiologia , Células Cultivadas , Feminino , Membro Posterior/fisiopatologia , Imunização Passiva , Atividade Motora , Regeneração Nervosa , Neuritos/efeitos dos fármacos , Neuritos/fisiologia , Coelhos , Ratos , Ratos Sprague-Dawley , Traumatismos da Medula Espinal/imunologia , Traumatismos da Medula Espinal/fisiopatologia , Tenascina/imunologia , Proteína rhoA de Ligação ao GTP/metabolismo
4.
Zhongguo Dang Dai Er Ke Za Zhi ; 14(5): 380-4, 2012 May.
Artigo em Chinês | MEDLINE | ID: mdl-22613112

RESUMO

OBJECTIVE: To study long-term behavioral and ultrastructural alterations in a hypoxic-ischemic brain damage (HIBD) model of neonatal rats. METHODS: Sixty seven-day-old Sprague-Dawley rats were randomly subjected to unilateral carotid artery ligation followed by hypoxic exposure (HIBD group) or sham operation (n=30 each). A battery of behavioral tests, including Morris water maze test and sensorimotor tests, were performed at a postnatal age of 5 weeks. Nissl staining was used for counting neurons. Transmission electron microscopy was used for observing synapse structures and measuring the thickness of the postsynaptic density area and the length of the postsynaptic active area. The correlations of histological changes with the results of behavioral tests were evaluated. RESULTS: The HIBD group showed a significantly longer escape latency (P<0.05) and a lower frequency of original platform crossing (P<0.05) in the Morris water maze test compared with the sham operation group. The sensorimotor function test showed that the sensorimotor function in the HIBD group was worse than in the sham operation group. Nissl staining showed that the number of neurons in the HIBD group was significantly reduced (P<0.01) compared with the sham operation group. Transmission electron microscopy showed that synapses were significantly reduced in number, and that the thickness of the postsynaptic density area and the length of the postsynaptic active area were reduced in the HIBD group. The thickness of the postsynaptic density area was negatively correlated with escape latency in the Morris water maze test (r=-0.861, P<0.01), and also negatively correlated with the total score of sensorimotor function tests (r=-0.758, P<0.05) in the HIBD group. CONCLUSIONS: Hypoxia ischemia can lead to neuron loss and ultrastructure damage, resulting in long-term deficit of behavioral functions in neonatal rats.


Assuntos
Encéfalo/ultraestrutura , Hipóxia-Isquemia Encefálica/psicologia , Animais , Animais Recém-Nascidos , Encéfalo/patologia , Feminino , Hipóxia-Isquemia Encefálica/patologia , Masculino , Aprendizagem em Labirinto , Microscopia Eletrônica de Transmissão , Ratos , Ratos Sprague-Dawley , Tempo de Reação
5.
Cancer Biol Ther ; 13(5): 341-8, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-22258034

RESUMO

Vasculogenic mimicry (VM), a process involving the formation of a tubular structure by highly invasive and genetically dysregulated tumor cells, can supplement the function of blood vessels to transport nutrients and oxygen to maintain the growth of tumor cells in many malignant tumors. We aimed to explore the existence of VM and its clinical significance in medulloblastoma in this study. VM was identified in 9 out of 41 (22%) medulloblastoma tissues. Immunohistochemical studies revealed that the presence of VM was associated with the expression of MMP-2, MMP-14, EphA2 and laminin 5γ2. Tumor tissues with VM were associated with lower microvessel density (MVD), which was indirect evidence of the blood supply function of VM. Survival analysis and log-rank tests showed that patients with VM had shorter overall survival time than those without VM. Multivariate analysis and the Cox proportional hazards model identified VM as independent prognostic factor for overall survival. Our results confirmed the existence of VM for the first time and revealed that VM is a strong independent prognostic factor for survival in patients with medulloblastoma.


Assuntos
Neoplasias Cerebelares/irrigação sanguínea , Meduloblastoma/irrigação sanguínea , Adolescente , Adulto , Neoplasias Cerebelares/metabolismo , Neoplasias Cerebelares/patologia , Criança , Pré-Escolar , Feminino , Humanos , Imuno-Histoquímica , Lactente , Masculino , Meduloblastoma/metabolismo , Meduloblastoma/patologia , Neovascularização Patológica/metabolismo , Neovascularização Patológica/patologia , Adulto Jovem
6.
Neurochem Res ; 36(12): 2391-400, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-21877237

RESUMO

Transdifferentiated and untransdifferentiated mesenchymal stem cells (MSCs) have shown therapeutic benefits in central nervous system (CNS) injury. However, it is unclear which would be more appropriate for transplantation. To address this question, we transplanted untransdifferentiated human umbilical mesenchymal stem cells (HUMSCs) and transdifferentiated HUMSCs (HUMSC-derived neurospheres, HUMSC-NSs) into a rat model of traumatic brain injury. Cognitive function, cell survival and differentiation, brain tissue morphology and neurotrophin expression were compared between groups. Significant improvements in cognitive function and brain tissue morphology were seen in the HUMSCs group compared with HUMSC-NSs group, which was accompanied by increased neurotrophin expression. Moreover, only few grafted cells survived in both the HUMSCs and HUMSC-NSs groups, with very few of the cells differentiating into neural-like cells. These findings indicate that HUMSCs are more appropriate for transplantation and their therapeutic benefits may be associated with neuroprotection rather than cell replacement.


Assuntos
Lesões Encefálicas/fisiopatologia , Lesões Encefálicas/cirurgia , Diferenciação Celular , Transdiferenciação Celular , Transplante de Células-Tronco Mesenquimais , Animais , Fator Neurotrófico Derivado do Encéfalo/biossíntese , Sobrevivência Celular , Cognição , Humanos , Aprendizagem em Labirinto , Células-Tronco Mesenquimais/fisiologia , Fatores de Crescimento Neural/biossíntese , Neurônios/citologia , Ratos , Ratos Sprague-Dawley
7.
Cancer Biol Ther ; 11(11): 974-80, 2011 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-21502808

RESUMO

Cancer stem cells are defined as a subpopulation of cancer cells with the capacity to self-renew and differentiate, which may play critical roles in tumor initiation, progress and resistance to current treatments. It has been reported that Dendritic cells (DCs) transfected with total tumor RNA could induce strong antitumor T-cell responses both in vivo and in vitro. In the study, we investigated the characteristics of 9L tumor spheres, and evaluated the antitumor effects of DCs transfected with 9L tumor spheres RNA in vivo. The results showed that 9L tumor spheres have the properties of cancer stem cells, and the majority of 9L cells were positive for CD133 and nestin. DCs transfected with 9L tumor spheres RNA can significantly inhibit glioma growth and prolong the survival of 9L glioma-bearing rats. These results demonstrated that 9L cancer stem like cells were enriched in tumor spheres, and they were a part of CD133+ cells, DCs transfected with cancer stem cells RNA may be an effective therapy for glioma.


Assuntos
Neoplasias Encefálicas/patologia , Vacinas Anticâncer/genética , Células Dendríticas/transplante , Gliossarcoma/patologia , Células-Tronco Neoplásicas/patologia , RNA/imunologia , Antígeno AC133 , Animais , Antígenos CD/metabolismo , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/terapia , Vacinas Anticâncer/uso terapêutico , Técnicas de Cultura de Células , Linhagem Celular Tumoral , Meios de Cultura , Células Dendríticas/metabolismo , Fator de Crescimento Epidérmico/farmacologia , Fator 2 de Crescimento de Fibroblastos/farmacologia , Gliossarcoma/genética , Gliossarcoma/terapia , Glicoproteínas/metabolismo , Interferon gama/sangue , Proteínas de Filamentos Intermediários/metabolismo , Estimativa de Kaplan-Meier , Células-Tronco Neoplásicas/efeitos dos fármacos , Células-Tronco Neoplásicas/imunologia , Proteínas do Tecido Nervoso/metabolismo , Nestina , Peptídeos/metabolismo , Ratos , Transfecção , Ensaios Antitumorais Modelo de Xenoenxerto
8.
Neurosci Bull ; 27(2): 83-90, 2011 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-21441969

RESUMO

OBJECTIVE: To investigate the role of tyrosine kinase receptor C (TrkC), the receptor of neurotrophin-3 (NT-3), in neuroplasticity following spinal cord injury (SCI). METHODS: Rats with cord transection were allowed to survive for 1, 3, 7 and 14 d post operation (dpo). TrkC expressions at lower thoracic levels of the spinal cord and in precentral gyrus of cerebral cortex were investigated. RESULTS: TrkC protein levels at both the site of injury (T10-T11) and the neighboring segments (T9 and T12) in the spinal cord decreased significantly at 1-7 dpo, followed by a rapid increase at 14 dpo. The temporal changes in TrkC mRNA expression level showed a similar pattern with that of TrkC protein. In addition, the levels of TrkC protein and mRNA at the site of injury (T10-T11) were significantly higher than those at the neighboring spinal segments (T9 and T12). Besides, the levels of TrkC protein and mRNA were higher at the rostral segment than at the caudal segment. However, in the motor cortex, TrkC protein was not detected and TrkC mRNA was expressed at a very low level. CONCLUSION: These results suggest that TrkC may be involved in neuroplasticity after SCI.


Assuntos
Córtex Cerebral/metabolismo , Regulação da Expressão Gênica/fisiologia , Receptor trkC/metabolismo , Traumatismos da Medula Espinal/patologia , Medula Espinal/metabolismo , Animais , Lateralidade Funcional/fisiologia , Proteína Glial Fibrilar Ácida/metabolismo , Indóis , Masculino , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Receptor trkC/genética , Medula Espinal/patologia , Fatores de Tempo
9.
Cytotherapy ; 13(1): 46-53, 2011 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-20735164

RESUMO

BACKGROUND AIMS: This study aimed to observe nine factors expressed in rat ischemic brain after transplantation of bone marrow stromal cells (BMSC) and/or endothelial progenitor cells (EPC). These factors were vascular endothelial growth factor (VEGF), stromal cell-derived factor-1 (SDF-1), basic fibroblast growth factor (bFGF), insulin-like growth factor (IGF-l), transforming growth factor-ß (TGF-ß), platelet-derived growth factor-BB (PDGF-BB), brain-derived neurotrophic factor (BDNF), glial cell line-derived neurotrophic factor (GDNF) and nerve growth factor (NGF). METHODS: Adult Wistar rats were divided randomly into four groups: a vehicle group, BMSC group, EPC group and BMSC combined with EPC group. The rats were subjected to middle cerebral artery occlusion (MCAO) then implanted intravenously with 3 × 10(6) BMSC, EPC, BMSC/EPC or phosphate-buffered saline (PBS) 24 h after MCAO. Neurologic functional deficits were measured on days 1, 7, 14, 28 after transplantation. On day 7 after transplantation, quantitative reverse transcription (qRT)-polymerase chain reaction (PCR) and Western blot were employed to detect the expression of VEGF, SDF-1, bFGF, IGF-l, TGF-ß, PDGF-BB, BDNF, GDNF and NGF. RESULTS: The neurologic evaluation found that the neurologic severity scores were no different between the four groups on day 1, and the scores of rats in the BMSC/EPC group were significantly lower than those of rats in the other groups on days 7, 14 and 28 after transplantation. The expressions of bFGF, VEGF and BNDF were significantly higher in the BMSC/EPC group compared with the other groups. CONCLUSIONS: The intravenous transplantation of BMSC combined with EPC could promote the functional rehabilitation of rats with focal cerebral ischemia, and the mechanism may be related to the enhanced expression of factors.


Assuntos
Células da Medula Óssea/citologia , Isquemia Encefálica/metabolismo , Encéfalo/metabolismo , Encéfalo/patologia , Citocinas/metabolismo , Células Endoteliais/transplante , Transplante de Células-Tronco , Animais , Comportamento Animal , Células da Medula Óssea/metabolismo , Isquemia Encefálica/patologia , Isquemia Encefálica/fisiopatologia , Isquemia Encefálica/terapia , Células Endoteliais/citologia , Microvasos/patologia , Fatores de Crescimento Neural/genética , Fatores de Crescimento Neural/metabolismo , Ratos , Ratos Wistar , Células-Tronco/citologia , Células-Tronco/metabolismo , Células Estromais/citologia , Células Estromais/transplante
10.
Nan Fang Yi Ke Da Xue Xue Bao ; 30(9): 2059-62, 2010 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-20855249

RESUMO

OBJECTIVE: To explore the possible mechanism of lipopolysaccharide (LPS)-induced cardiomyocyte hypertrophy in rats. METHODS: Neonatal rat cardiomyocytes cultured in vitro were stimulated with 100 µg/L LPS for 1, 4 or 8 h and scanned by atomic force microscopy (AFM) for measurement of the two-dimensional area, three-dimensional surface area and volume of each cell. The total proteins and Na(+)-K(+)-ATPase activity in the cardiomyocytes were determined. The same measurements were also carried out in neonatal rat cardiomyocyte cultures stimulated by 0.5 µmol/L ouabain for 8 h and the total protein levels were measured. RESULTS: Following a 8-hour stimulation with LPS, the two-dimensional area, three-dimensional surface area and volume of the single cardiomyocyte became enlarged and the total cellular proteins increased significantly as compared with those in the normal control cells (P < 0.05). LPS treatment for 4 and 8 h resulted in significantly decreased activity of Na(+)-K(+)-ATPase in the cardiomyocytes (P < 0.05). In the cells treated with ouabain for 8 h, the two-dimensional area, three-dimensional surface area, volume of the single cardiomyocyte and the total cellular proteins increased significantly in comparison with the normal control group (P < 0.05). CONCLUSION: LPS can result in cardiomyocyte hypertrophy in rats possibly in relation to lowered Na(+)-K(+)-ATPase activity in the cardiomyocytes after LPS exposure.


Assuntos
Crescimento Celular/efeitos dos fármacos , Miócitos Cardíacos/patologia , ATPase Trocadora de Sódio-Potássio/metabolismo , Animais , Animais Recém-Nascidos , Células Cultivadas , Hipertrofia/induzido quimicamente , Lipopolissacarídeos , Miócitos Cardíacos/enzimologia , Ratos , Ratos Wistar
11.
Neurochem Res ; 35(10): 1522-9, 2010 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-20658188

RESUMO

Mesenchymal stem cells are capable of differentiating into dopaminergic-like cells, but currently no report has been available to describe the induction of human umbilical vein mesenchymal stem cells (HUVMSCs) into dopaminergic-like cells. In this study, we induced HUVMSCs in vitro into neurospheres constituted by neural stem-like cells, and further into cells bearing strong morphological, phenotypic and functional resemblances with dopaminergic-like cells. These HUVMSC-derived dopaminergic-like cells, after grafting into the brain of a rat model of Parkinson's disease (PD), showed a partial therapeutic effect in terms of the behavioral improvement. Nerve growth factor was reported to improve the local microenvironment of the grafted cells, and we therefore further tested the effect of dopaminergic-like cell grafting combined with nerve growth factor (NGF) administration at the site of cell transplantation. The results showed that NGF administration significantly promoted the survival of the grafted cells in the host brain and enhanced the content of dopaminergic in the local brain tissue. Behavioral test demonstrated a significant improvement of the motor function of the PD rats after dopaminergic-like cell grafting with NGF administration as compared with that of rats receiving the cell grafting only. These results suggest that transplantation of the dopaminergic-like cells combined with NGF administration may represent a new strategy of stem cell therapy for PD.


Assuntos
Dopamina/metabolismo , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/metabolismo , Atividade Motora , Fator de Crescimento Neural/uso terapêutico , Doença de Parkinson/terapia , Veias Umbilicais/citologia , Animais , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Encéfalo/patologia , Diferenciação Celular , Humanos , Células-Tronco Mesenquimais/citologia , Doença de Parkinson/metabolismo , Doença de Parkinson/psicologia , Ratos , Ratos Sprague-Dawley
12.
Neuroimmunomodulation ; 17(4): 270-8, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-20203533

RESUMO

LINGO-1 (leucine-rich repeat and Ig domain-containing, Nogo receptor-interacting protein) is an important component of the NgR receptor complex involved in RhoA activation and axon regeneration. The authors report on passive immunization with LINGO-1 polyclonal antiserum, a therapeutic approach to overcome NgR-mediated growth inhibition after spinal cord injury (SCI). The intrathecally administered high-titer rabbit-derived antiserum can be detected around the injury site within a wide time window; it blocks LINGO-1 in vivo with high molecular specificity. In this animal model, passive immunization with LINGO-1 antiserum significantly decreased RhoA activation and increased neuronal survival. Adult rats immunized in this manner show recovery of certain hindlimb motor functions after dorsal hemisection of the spinal cord. Thus, passive immunotherapy with LINGO-1 polyclonal antiserum may represent a promising repair strategy following acute SCI.


Assuntos
Citoproteção/efeitos dos fármacos , Imunização Passiva/métodos , Proteínas de Membrana/antagonistas & inibidores , Proteínas do Tecido Nervoso/antagonistas & inibidores , Recuperação de Função Fisiológica/efeitos dos fármacos , Traumatismos da Medula Espinal/tratamento farmacológico , Animais , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/imunologia , Citoproteção/imunologia , Modelos Animais de Doenças , Feminino , Soros Imunes/imunologia , Soros Imunes/farmacologia , Injeções Espinhais , Proteínas de Membrana/imunologia , Degeneração Neural/tratamento farmacológico , Degeneração Neural/imunologia , Degeneração Neural/fisiopatologia , Proteínas do Tecido Nervoso/imunologia , Paralisia/tratamento farmacológico , Paralisia/imunologia , Paralisia/fisiopatologia , Ratos , Ratos Sprague-Dawley , Recuperação de Função Fisiológica/imunologia , Traumatismos da Medula Espinal/imunologia , Traumatismos da Medula Espinal/fisiopatologia , Resultado do Tratamento , Proteína rhoA de Ligação ao GTP/efeitos dos fármacos , Proteína rhoA de Ligação ao GTP/metabolismo
13.
Int J Cancer ; 127(9): 2222-9, 2010 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-20127864

RESUMO

Inhibition of tumor neovascularization has profound effects on the growth of solid tumors. Our previous studies have shown the effect of VEGF165-PE38 recombinant immunotoxin on proliferation and apoptosis in human umbilical vein endothelial cells in vitro. In this study, we explored the direct inhibition of angiogenesis in chick chorioallantoic membrane and antiangiogenic therapy in a malignant glioma model. HEK293 cells were transfected with the pVEGF165PE38-IRES2-EGFP plasmid. ELISA was used to confirm the expression of VEGF165-PE38 in the transfected cells. These cells released 1396 + or - 131.9 pg VEGF165-PE38/1x10(4) cells/48 h into the culture medium and the supernatant was capable of inhibiting the growth of capillary-like structures in chick chorioallantoic membrane assay. In a murine malignant glioma model, plasmid was directly administered via multiple local intratumoral delivery. After day 16 the tumor volume in mice treated with pVEGF165PE38-IRES2-EGFP was significantly lower than that in mice in the control groups. Immunohistochemistry studies showed that the treated group had decreased expression of CD31. Quantitative analysis of microvessel density in the treated group was 1.99 + or - 0.69/0.74 mm(2), and was significantly lower than that in the control groups (9.33 + or - 1.99/0.74 mm(2), 8.09 + or - 1.39/0.74 mm(2) and 8.49 + or - 1.69/0.74 mm(2)). Immunohistochemistry analysis indicated that immunotoxin VEGF165-PE38 was distributed in the treated group in malignant glioma tissue. Our findings provide evidence that the in vivo production of VEGF165-PE38 through gene therapy using a eukaryotic expression plasmid had potential antiangiogenic activity in malignant glioma in vivo.


Assuntos
Inibidores da Angiogênese/uso terapêutico , Terapia Genética , Glioma/terapia , Imunotoxinas/uso terapêutico , Fator A de Crescimento do Endotélio Vascular/genética , ADP Ribose Transferases/uso terapêutico , Animais , Toxinas Bacterianas/uso terapêutico , Linhagem Celular Tumoral , Exotoxinas/uso terapêutico , Estudos de Viabilidade , Glioma/irrigação sanguínea , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Plasmídeos , Pseudomonas/metabolismo , Transfecção , Fatores de Virulência/uso terapêutico , Ensaios Antitumorais Modelo de Xenoenxerto , Exotoxina A de Pseudomonas aeruginosa
14.
Cell Mol Neurobiol ; 30(2): 275-82, 2010 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-19757023

RESUMO

Myelin-derived proteins, such as tenascin-R (TN-R), myelin associate glycoprotein (MAG), oligodendrocyte-myelin glycoprotein (OMgp), and Nogo-A, inhibit the central nervous system regeneration. In this study, the DNA vaccine encoding for oligodendrocyte and myelin-related antigens was employed to attenuate the axonal growth inhibitory properties of myelin in the setting of spinal cord injury. Using a rat spinal cord dorsal hemisection model, the vaccine directed against the inhibitory epitopes of Nogo-A, MAG, OMgp, and TN-R was administered intramuscularly once a week following spinal cord injury, supplemented with local application of specific anti-sera against the four antigens. Anterograde labeling of dorsal column fibers showed active axonal regeneration through the lesion site at the eighth week following the treatment in experimental group but not in control groups. Light microscopic and ultrastructural analysis revealed that vaccination with these myelin-related antigens did not lead to demyelinating disease. OMgp and TN-R levels were down-regulated at the lesion site together with a parallel increase in growth-associated protein 43 levels in the treatment groups. This study reveals the effective approach of a DNA vaccine strategy by attaining the special antibody to direct neutralization of the myelin inhibitors during spinal cord injury.


Assuntos
Axônios/fisiologia , Traumatismos da Medula Espinal/terapia , Vacinas de DNA/uso terapêutico , Animais , Axônios/ultraestrutura , Encefalomielite Autoimune Experimental/patologia , Epitopos/genética , Feminino , Proteínas Ligadas por GPI , Imunização Passiva , Atividade Motora , Proteínas da Mielina/genética , Proteínas da Mielina/imunologia , Glicoproteína Associada a Mielina/genética , Glicoproteína Associada a Mielina/imunologia , Glicoproteína Associada a Mielina/metabolismo , Glicoproteína Mielina-Oligodendrócito , Regeneração Nervosa , Proteínas Nogo , Ratos , Ratos Endogâmicos Lew , Medula Espinal/patologia , Traumatismos da Medula Espinal/patologia , Tenascina/genética , Tenascina/imunologia , Tenascina/metabolismo
15.
Differentiation ; 79(1): 15-20, 2010 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-19800163

RESUMO

Human Wharton's jelly-derived mesenchymal stromal cells (hWJ-MSCs) are capable of differentiating into neural and astroglia-like cell types. However, a reliable means of inducing the selective differentiation of hWJ-MSCs into oligodendrocyte progenitor cells (OPCs) in vitro has not yet been established. In this study, the OPC-like differentiation of hWJ-MSCs was characterized using and immunoblotting. The hWJ-MSC-derived OPC-like cells were able to secrete nerve growth factors and promote neurite outgrowth in vitro. These results show that hWJ-MSCs can be induced to differentiate into cells with the morphologic, phenotypic and functional characteristics of OPC-like cells.


Assuntos
Diferenciação Celular/fisiologia , Células-Tronco Mesenquimais/citologia , Fator de Crescimento Neural/metabolismo , Neurônios/citologia , Oligodendroglia/citologia , Células-Tronco/citologia , Cordão Umbilical/citologia , Separação Celular , Células Cultivadas , Feminino , Humanos , Imunofenotipagem , Neurônios/metabolismo , Oligodendroglia/metabolismo , Gravidez , Células-Tronco/metabolismo , Células Estromais/metabolismo
16.
Nan Fang Yi Ke Da Xue Xue Bao ; 29(11): 2175-8, 2009 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-19923059

RESUMO

OBJECTIVE: To express and purify the fusion protein of extracellular domain of human Ig domain-containing, neurite outgrowth inhibitor (Nogo) receptor-interacting protein-1 (LINGO-1(aa76-319)) in prokaryotic cells and prepare the rabbit anti-LINGO-1 polyclonal antibody (pAb). METHODS: The 732 bp DNA sequence of hLINGO-1(aa76-319) was obtained from pCMV-SPORT6 by PCR and inserted into pET30a(+) plasmid to construct the prokaryotic expression plasmid pET30a(+)-hLINGO-1(aa76-319), which was subsequently transformed into E.coli. The target fusion protein was expressed with IPTG induction and purified by Ni(2+)-NTA affinity chromatography column. The antiserum against hLINGO-1(aa76-319) was obtained from the rabbits immunized with hLINGO-1(aa76-319), and the titer of the pAb was determined using enzyme linked immunosorbent assay (ELISA) and its specificity identified using Western blotting. RESULTS: The prokaryotic expression plasmid pET30a(+)-hLINGO-1(aa76-319) was constructed successfully. Efficient expression of the target fusion protein was achieved with IPTG induction at the optimal concentration of 0.4 mmol/L and culture temperature at 37 degrees celsius; for 2.5 h. The hLINGO-1(aa76-319) fusion protein was effectively expressed in E.coli as inclusion bodies, and the soluble protein was obtained through denaturation and refolding procedures, and the purified fusion protein showed a purity above 90%. The titer of the anti-hLINGO-1(aa76-319) pAb obtained by immunizing the rabbits with the purified protein reached 1:1.6x10(6), and Western blotting confirmed its good specificity. CONCLUSION: The fusion protein hLINGO-1(aa76-319) with high purity has been obtained and the anti-hLINGO-1(aa76-319) pAb obtained shows a high titer and good specificity, which provide important experimental basis for further functional investigation of LINGO-1.


Assuntos
Anticorpos/isolamento & purificação , Proteínas de Membrana/biossíntese , Proteínas de Membrana/imunologia , Proteínas do Tecido Nervoso/biossíntese , Proteínas do Tecido Nervoso/imunologia , Proteínas Recombinantes de Fusão/imunologia , Animais , Anticorpos/imunologia , Especificidade de Anticorpos , Escherichia coli/genética , Escherichia coli/metabolismo , Humanos , Soros Imunes/imunologia , Proteínas de Membrana/genética , Proteínas do Tecido Nervoso/genética , Plasmídeos/genética , Coelhos , Proteínas Recombinantes de Fusão/biossíntese , Proteínas Recombinantes de Fusão/genética
17.
J Neurosci Methods ; 179(1): 45-50, 2009 Apr 30.
Artigo em Inglês | MEDLINE | ID: mdl-19428510

RESUMO

Bone marrow stroma cells-derived neural stem cells (BMSCs-D-NSCs) transplantation is a promising strategy for the treatment of nervous system disorders. The development of a non-invasive method to follow the fate of BMSCs-D-NSCs in vivo is very important for the future application of this treatment. In this paper, we show for the first time, that BMSCs-D-NSCs from rhesus monkeys can be labeled in vitro with the superparamagnetic iron oxide (SPIO) contrast agent Feridex and Poly-L-lysine (PLL) without affecting morphology, cell cycle, telomerase activity, proliferation and differentiation ability of the labeled cells. Furthermore, when autografted into the striatum, these cells survived, differentiated and were incorporated into the brain, and could be reliably tracked using MRI, as confirmed by histological examination of the grafting sites with PKH(67) fluorescence. These results suggest that Feridex labeling of BMSCs-D-NSCs is feasible, efficient and safe for MRI tracing following autografting into the rhesus monkey nervous system.


Assuntos
Células da Medula Óssea/citologia , Encéfalo/citologia , Imageamento por Ressonância Magnética/métodos , Neurônios/fisiologia , Transplante de Células-Tronco , Células-Tronco/fisiologia , Animais , Apoptose , Células da Medula Óssea/ultraestrutura , Encéfalo/ultraestrutura , Ciclo Celular , Sobrevivência Celular , Dextranos , Óxido Ferroso-Férrico , Imuno-Histoquímica , Ferro , Macaca mulatta , Nanopartículas de Magnetita , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Neurônios/citologia , Neurônios/ultraestrutura , Óxidos , Células-Tronco/citologia , Células-Tronco/ultraestrutura , Células Estromais/fisiologia , Células Estromais/ultraestrutura , Telomerase/metabolismo , Transplante Autólogo
18.
Neurosci Lett ; 458(3): 116-21, 2009 Jul 24.
Artigo em Inglês | MEDLINE | ID: mdl-19394407

RESUMO

This study is designed to evaluate the therapeutic effects of three types of neurospheres (NSs) derived from brain, bone marrow and adipose tissue in a rat model of spinal contusive injury. As shown by BBB locomotor rating scale and grid test, the optimal therapeutic responses generated by subventricular zone-derived NSs (SVZ-NSs), and followed by adipose-derived (AD-NSs) and bone marrow-derived NSs (BM-NSs) after being grafted into the injured spinal cord. In three cell-treated groups, very few (<1%) grafted cells survived and these survived cells mainly differentiated into oligodendrocytes at week 12 after injury. Additionally, all the cell-treated groups, especially in the SVZ-treated group showed an increase in host oligodendrocytes than control group. Moreover, the level of selective neurotrophins (NTs) in the SVZ-NSs group were significantly higher than those in the BM-NSs and AD-NSs groups, and the level of NTs in the saline group was also significantly higher than sham group. Therefore, not cell replacement or infusion but neuroprotective action associated with endogenous oligodendrocytes and NTs that active by the grafted NSs may contribute to the functional recovery.


Assuntos
Transplante de Medula Óssea , Neurônios/transplante , Traumatismos da Medula Espinal/cirurgia , Transplante de Células-Tronco , Tecido Adiposo/transplante , Animais , Masculino , Fatores de Crescimento Neural/metabolismo , Regeneração Nervosa , Oligodendroglia/fisiologia , Ratos , Ratos Sprague-Dawley , Recuperação de Função Fisiológica , Traumatismos da Medula Espinal/fisiopatologia
19.
Biotechnol Lett ; 31(2): 181-9, 2009 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-18923908

RESUMO

Human mesenchymal stem cells-like cells (hMSCs-like cells) were used as a tumor treatment platform for the systemic delivery of immunotoxin genes. VEGF165-PE38 recombinant immunotoxin served as the model system. hMSCs-like cells were isolated, expanded, and electroporated with the pIRES2-VEGF165PE38-EGFP plasmid. RT-PCR and ELISA were used to confirm the expression of VEGF165-PE38 in the transfected hMSCs-like cells. These cells released 1390 +/- 137 pg VEGF165-PE38/10(4)cells over 48 h into the culture medium and the supernatant was capable of selectively killing human umbilical vein endothelial cells (HUVECs) and increasing apoptosis in these cells. In contrast, RPMI8226 was not inhibited by identical supernatants. Thus, these results lay the foundation for further studies on the potential role of hMSCs-like cells as a targeted therapeutic delivery vehicle for immunotoxins.


Assuntos
Células Endoteliais/citologia , Células Endoteliais/imunologia , Imunotoxinas/imunologia , Células-Tronco Mesenquimais/imunologia , Fator A de Crescimento do Endotélio Vascular/imunologia , Comunicação Celular/imunologia , Sobrevivência Celular , Células Cultivadas , Humanos , Imunotoxinas/administração & dosagem , Transfecção , Fator A de Crescimento do Endotélio Vascular/administração & dosagem , Fator A de Crescimento do Endotélio Vascular/genética
20.
Cell Biol Int ; 33(4): 466-74, 2009 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-18725309

RESUMO

Glioma is the most common primary intracranial malignant tumor. Despite advances in surgical techniques and adjuvant radio- and chemotherapies, the prognosis for patients with glioma remains poor. We have explored the effects of using genetically modified mesenchymal stem cells (MSCs) to treat malignant glioma in rats. Mesenchymal stem cells isolated from Sprague-Dawley rats can directly suppress the growth of C6 cells in vitro. MSCs transplanted intratumorally can also significantly inhibit the growth of glioma and prolong survival in C6 glioma-bearing models. MSCs producing Interleukin-18 infected by adenoviral vector inhibited glioma growth and prolonged the survival of glioma-bearing rats. Transplantation of IL-18 secreting MSCs was associated with enhanced T cell infiltration and long-term anti-tumor immunity. Thus, IL-18 may be an effective adoptive immunotherapy for malignant glioma. When used in conjunction with MSCs as targeting vehicles in vivo, IL-18 may offer a promising new treatment option for malignant glioma.


Assuntos
Neoplasias Encefálicas/terapia , Terapia Genética , Glioma/terapia , Interleucina-18/genética , Transplante de Células-Tronco Mesenquimais , Animais , Neoplasias Encefálicas/diagnóstico por imagem , Neoplasias Encefálicas/patologia , Linhagem Celular Tumoral , Vetores Genéticos , Glioma/diagnóstico por imagem , Glioma/patologia , Estimativa de Kaplan-Meier , Linfócitos do Interstício Tumoral/metabolismo , Células-Tronco Mesenquimais/metabolismo , Camundongos , Radiografia , Ratos , Ratos Sprague-Dawley
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