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1.
Sci Rep ; 9(1): 18321, 2019 Dec 04.
Artigo em Inglês | MEDLINE | ID: mdl-31797961

RESUMO

The novel methods for efficient plant regeneration via direct somatic embryogenesis (SE) and SE-mediated transformation system under high concentration of NAA in Ranunculus sceleratus were established. On MS media containing a high concentration of NAA (10.0 mg/L) in the dark, all inoculated explants (root, stem and leaf) formed somatic embryos at high frequencies, respectively, 66.03, 126.47 and 213.63 embryoids per explant, and 100% of the embryoids developed into plantlets on 1/2 MS rooting media. Morphological and histological analyses revealed that SE in R. sceleratus followed a classical pattern. All inoculated explants can be used as receptors for genetic transformation in R. sceleratus, through direct SE-mediated method after Agrobacterium infection. RcLEC1-B, as a marker gene, changed the number and morphology of flower organs and the development of cuticle in R. sceleratus, which indicated that the efficient transgenic system of R. sceleratus was established. To our knowledge, this is the first observation that both direct SE and transgenic transformation system, via induction of a single plant growth regulator, have been successfully constructed in R. sceleratus.

2.
Brain Res ; 1657: 279-287, 2017 02 15.
Artigo em Inglês | MEDLINE | ID: mdl-28011395

RESUMO

Salvianolic acid A (Sal A), a bioactive compound isolated from the Chinese medicinal herb Danshen, is used for the prevention and treatment of cardiovascular diseases. However, the protective function of Sal A on preserving the role of blood-spinal cord barrier (BSCB) after spinal cord injury (SCI) is unclear. The present study investigated the effects and mechanisms of Sal A (2.5, 5, 10mg/kg, i.p.) on BSCB permeability at different time-points after compressive SCI in rats. Compared to the SCI group, treatment with Sal A decreased the content of the Evans blue in the spinal cord tissue at 24h post-SCI. The expression levels of tight junction proteins and HO-1 were remarkably increased, and that of p-caveolin-1 protein was greatly decreased after SCI Sal A. The effect of Sal A on the expression level of ZO-1, occluding, and p-caveolin-1 after SCI was blocked by the HO-1 inhibitor, zinc protoporphyrin IX (ZnPP). Also, Sal A inhibited the level of apoptosis-related proteins and improved the motor function until 21days after SCI. In addition, Sal A significantly increased the expression of microRNA-101 (miR-101) in the RBMECs under hypoxia. AntagomiR-101 markedly increased the RBMECs permeability and the expression of the Cul3 protein by targeting with 3'-UTR of its mRNA. The expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and HO-1 was significantly increased after agomiR-101 treatment. Therefore, Sal A could improve the recovery of neurological function after SCI, which could be correlated with the repair of BSCB integrity by the miR-101/Cul3/Nrf2/HO-1 signaling pathway.


Assuntos
Ácidos Cafeicos/farmacologia , Permeabilidade Capilar/efeitos dos fármacos , Fármacos Cardiovasculares/farmacologia , Lactatos/farmacologia , Traumatismos da Medula Espinal/tratamento farmacológico , Medula Espinal/irrigação sanguínea , Medula Espinal/efeitos dos fármacos , Animais , Permeabilidade Capilar/fisiologia , Caveolina 1/metabolismo , Proteínas Culina/metabolismo , Modelos Animais de Doenças , Avaliação Pré-Clínica de Medicamentos , Heme Oxigenase (Desciclizante)/metabolismo , Hipóxia/tratamento farmacológico , Hipóxia/metabolismo , Masculino , MicroRNAs/metabolismo , Atividade Motora/efeitos dos fármacos , Atividade Motora/fisiologia , Fator 2 Relacionado a NF-E2/metabolismo , Fármacos Neuroprotetores/farmacologia , Distribuição Aleatória , Ratos Sprague-Dawley , Medula Espinal/metabolismo , Traumatismos da Medula Espinal/metabolismo , Junções Íntimas/efeitos dos fármacos , Junções Íntimas/metabolismo , Proteína da Zônula de Oclusão-1/metabolismo
3.
Neurosci Lett ; 604: 18-23, 2015 Sep 14.
Artigo em Inglês | MEDLINE | ID: mdl-26079325

RESUMO

Acetyl-l-carnitine (ALC) facilitates the entry and exit of fatty acids from mitochondria and plays an essential role in energy metabolism. Although ALC is known to exert neuroprotective effects in multiple neurological diseases, its effects on spinal cord injury (SCI)-induced mitochondrial impairments and apoptosis remain unclear. In this study, we aimed to evaluate the putative effects of ALC on mitochondrial dysfunction and apoptosis induced by SCI in a rodent model. Our results indicate that SCI elicits dynamic alternations in the expression of mitochondria-related proteins. Transmission electron microscopy analysis showed that ALC administration abrogated key ultrastructural abnormalities in mitochondria at 24h after SCI by maintaining mitochondrial length, reducing the number of damaged mitochondria, and reversing mitochondrial score (P<0.05 compared with SCI group). In addition, ALC administration maintained the mitochondrial membrane potential and mitochondrial Na(+)-K(+)-ATPase activity following SCI (P<0.05 compared with SCI group). ALC administration reversed the downregulation of mitofusin 1 (Mfn1), Mfn2, Bcl-2, and the upregulation of dynamin-related protein 1 (Drp1), mitochondrial fission 1 (Fis1), Bcl-2-associated X protein (Bax) and cytosol cytochrome c (cyto-CytC) induced by SCI (P<0.05 compared with SCI group). Finally ALC administration greatly reduced the percentage of apoptotic cells compared with the SCI group (P<0.01). In conclusion, our findings demonstrated that ALC ameliorated SCI-induced mitochondrial structural alternations, mitochondrial dysfunction, and apoptosis.


Assuntos
Acetilcarnitina/metabolismo , Apoptose , Mitocôndrias/fisiologia , Traumatismos da Medula Espinal/metabolismo , Acetilcarnitina/farmacologia , Animais , Potencial da Membrana Mitocondrial , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/ultraestrutura , Proteínas Mitocondriais/metabolismo , Ratos , ATPase Trocadora de Sódio-Potássio/metabolismo , Traumatismos da Medula Espinal/patologia
4.
J Mol Neurosci ; 56(2): 388-96, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-26007330

RESUMO

This study was performed to investigate the effect of bone marrow stromal cells (BMSCs) combined with green tea polyphenols (GTPs) on the blood-spinal cord barrier (BSCB) permeability after spinal cord injury (SCI) in the rat model. In the model of SCI rats, we found that the water content and the BSCB permeability were decreased by BMSCs and GTPs treatment, and their combination had a synergistic effect. Further, the motor function of rats was also greatly improved by BMSCs and GTPs administration. After treated by the combination of BMSCs and GTPs, SCI rats showed the up-regulated expression of tight junction (TJ) associated proteins claudin-5, occludin and ZO-1 by Western blot, which was more remarkable than that in the single treatment. The increased expression levels of claudin-5, occludin, and ZO-1 were the most obvious in the spinal cord microvessels using immunohistochemistry assay. This led to the conclusion that the combination of BMSCs and GTPs could decrease the BSCB permeability by up-regulating protein expression levels of claudin-5, occludin, and ZO-1. In addition, after BMSCs and GTPs administration, the results of Western blot and enzyme-linked immunosorbent assay (ELISA) revealed a significant decrease in protein expression level and the activation of nuclear factor-кB (NF-кB) p65. Our results indicated that combination of BMSCs and GTPs could improve motor function after SCI, which might be correlated with improvements in BSCB integrity, and that NF-кB might be involved in the modulating process.


Assuntos
Permeabilidade Capilar , Transplante de Células-Tronco Mesenquimais , Extratos Vegetais/farmacologia , Polifenóis/farmacologia , Compressão da Medula Espinal/terapia , Medula Espinal/irrigação sanguínea , Animais , Células Cultivadas , Claudina-1/genética , Claudina-1/metabolismo , Masculino , NF-kappa B/metabolismo , Ocludina/genética , Ocludina/metabolismo , Extratos Vegetais/uso terapêutico , Polifenóis/uso terapêutico , Ratos , Ratos Sprague-Dawley , Medula Espinal/efeitos dos fármacos , Medula Espinal/metabolismo , Compressão da Medula Espinal/tratamento farmacológico , Chá/química , Regulação para Cima , Proteína da Zônula de Oclusão-1/genética , Proteína da Zônula de Oclusão-1/metabolismo
5.
Sci Rep ; 5: 8823, 2015 Mar 06.
Artigo em Inglês | MEDLINE | ID: mdl-25744384

RESUMO

A new approach was established for the regeneration of Trichosanthes kirilowii from root, stem, and leaf explants by somatic embryogenesis (SE), involving a previously unreported SE structure, rhizoid tubers (RTBs). During SE, special rhizoids were first induced from root, stem, and leaf explants with average rhizoid numbers of 62.33, 40.17, and 11.53 per explant, respectively, on Murashige and Skoog (MS) medium (pH 4.0) supplemented with 1.0 mg/L 1-naphthaleneacetic acid (NAA) under dark conditions. Further, one RTB was formed from each of the rhizoids on MS medium (pH 4.0) supplemented with 20 mg/L thidiazuron (TDZ) under light conditions. In the suitable range (pH 4.0-9.0), a lower pH value increased the induction of rhizoids and RTBs. Approximately 37.77, 33.47, and 31.07% of in vivo RTBs from root, stem, and leaf explants, respectively, spontaneously developed into multiple plantlets on the same MS medium (supplemented with 20 mg/L TDZ) for induction of RTBs, whereas >95.00% of in vitro RTBs from each kind of explant developed into multiple plantlets on MS medium supplemented with 5.0 mg/L 6-benzylaminopurine (BAP). Morphological and histological analyses revealed that RTB is a novel type of SE structure that develops from the cortex cells of rhizoids.


Assuntos
Concentração de Íons de Hidrogênio , Regeneração , Trichosanthes/fisiologia , Fenótipo , Folhas de Planta/crescimento & desenvolvimento , Brotos de Planta/crescimento & desenvolvimento , Caules de Planta/crescimento & desenvolvimento , Trichosanthes/crescimento & desenvolvimento
6.
J Neurol Sci ; 346(1-2): 51-9, 2014 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-25129208

RESUMO

Previous studies have shown that curcumin (Cur) can produce potent neuroprotective effects against damage due to spinal cord injury (SCI). However, whether Cur can preserve the function of the blood-spinal cord barrier (BSCB) is unclear. The present study was performed to investigate the mechanism underlying BSCB permeability changes, which were induced by treatment with Cur (75, 150, and 300 mg/kg, i.p.) after compressive SCI in rats. BSCB permeability was evaluated by Evans blue leakage. Motor recovery of rats with SCI was assessed using the Basso, Beattie, and Bresnahan scoring system every day until the 21st days post-injury. The protein levels of heme oxygenase-1 (HO-1), tight junction protein, and inflammatory factors were analyzed by western blots. The expression of the inflammatory factors tumor necrosis factor-α (TNF-α) and nuclear factor-kappaB (NF-κB) mRNA was determined with reverse transcription-polymerase chain reactions. Treatment with Cur (150 and 300 mg/kg) significantly reduced Evans blue leakage into the spinal cord tissue at 24h after SCI. Cur (150 mg/kg) significantly increased HO-1 protein expression. The levels of TNF-α and NF-κB mRNA and protein greatly increased at 24h after SCI, and this increase was significantly attenuated by Cur treatment. ZO-1 and occludin expression was upregulated by Cur (150 mg/kg) treatment after SCI, and this effect was blocked by the HO-1 inhibitor zinc protoporphyrin. Long-term effects of Cur on motor recovery after SCI were observed. Our results indicated that Cur can improve motor function after SCI, which could correlate with improvements in BSCB integrity.


Assuntos
Anti-Inflamatórios não Esteroides/uso terapêutico , Permeabilidade Capilar/efeitos dos fármacos , Curcumina/uso terapêutico , Compressão da Medula Espinal/tratamento farmacológico , Compressão da Medula Espinal/patologia , Junções Íntimas/efeitos dos fármacos , Animais , Modelos Animais de Doenças , Método Duplo-Cego , Azul Evans , Regulação da Expressão Gênica/efeitos dos fármacos , Masculino , Transtornos dos Movimentos/tratamento farmacológico , Transtornos dos Movimentos/etiologia , NF-kappa B/genética , NF-kappa B/metabolismo , Ocludina/genética , Ocludina/metabolismo , Protoporfirinas/uso terapêutico , Ratos , Ratos Sprague-Dawley , Medula Espinal/patologia , Compressão da Medula Espinal/complicações , Junções Íntimas/patologia , Fatores de Tempo , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo , Proteína da Zônula de Oclusão-1/genética , Proteína da Zônula de Oclusão-1/metabolismo
7.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 44(5): 747-51, 755, 2013 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-24325104

RESUMO

OBJECTIVE: To study the clinical effects of hypoxia preconditioning (HPC) and its effects on serum neuroglobin (NGB) and S-100B level in the patients undergoing intracranial aneurysm surgery. METHODS: Forty patients scheduled to intracranial aneurysm surgery were randomly.divided into 2 groups: HPC group (n= 20) and control group (n= 20). The patients in HPC group were treated with 3 cycles of deoxidation-reoxygenation after intubation. The time of deoxidation in each HPC cycle was recorded, while vital signs were also recorded in each corresponding time point. Blood samples were obtained from exsanguinate radial artery and jugular bulb section at the end of each HPC cycle and corresponding time points during operation to measure serum level of NGB and S100B protein and to analysis blood gas. RESULTS: During HPC process, the patients in group HPC experienced mild hypoxia and CO2 retention. With the times of HPC increasing, CO2 retention degree became heavier (P<0. 05) while hypoxia improved, the patients need more time to make SpO2 from 100% to 90% (P<0. 05). From T2 to T4 (the end of the third reoxygenation, during skull opened and aneurysm dipped, skull closed), NGB in group HPC was higher than that in control (P<0. 05), but S-100B level was not different between HPC and control group (P>0. 05). CONCLUSION: HPC could induce compensatory ability of the body to hypoxia, which might be related to the up-regulation of NGB expression.


Assuntos
Aneurisma Intracraniano/cirurgia , Precondicionamento Isquêmico/métodos , Proteínas do Tecido Nervoso/sangue , Proteínas S100/sangue , Adolescente , Adulto , Feminino , Globinas , Humanos , Aneurisma Intracraniano/sangue , Masculino , Pessoa de Meia-Idade , Neuroglobina , Regulação para Cima , Adulto Jovem
8.
J Mol Neurosci ; 51(3): 986-93, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-23943397

RESUMO

Salvianolic acid B (Sal B), a bioactive compound isolated from the Chinese medicinal herb danshen, is commonly used for the prevention and treatment of cardiovascular disease. The present study was performed to investigate the effect of Sal B on the blood-spinal cord barrier (BSCB) after spinal cord injury (SCI) in a rat model. Sal B (1, 10, and 50 mg/kg i.v.) was administered to rats immediately following SCI. The permeability of the BSCB and spinal cord tissue water content were evaluated. Additionally, the expression levels of tight junction proteins and heme oxygenase-1 (HO-1) were monitored by Western blot analysis. Enzyme-linked immunosorbent assay analysis of spinal cord tissue homogenates was performed 24 h post-SCI to evaluate the expression of inflammation-related cytokines. In addition, the motor recovery of SCI rats was assessed using the Basso, Beattie, and Bresnahan scoring system. Compared to the SCI group, rats treated with Sal B (10, 50 mg/kg) exhibited significantly reduced spinal cord tissue water content and BSCB permeability. Further, the motor function of rats was also greatly improved by Sal B administration. The expression of pro-inflammatory factors TNF-α and NF-κB was found to be greatly increased 24 h post-SCI, and this upregulation was significantly attenuated by Sal B treatment. The expression of ZO-1 and occludin was upregulated by Sal B (10 mg/kg) treatment after SCI, and this effect was blocked by the HO-1 inhibitor ZnPP. Taken together, our results clearly indicate that Sal B attenuates SCI by promoting the repair of the damaged BSCB, demonstrating that this molecule is a novel and promising therapeutic agent for human SCI.


Assuntos
Alcenos/farmacologia , Permeabilidade Capilar , Fármacos Neuroprotetores/farmacologia , Polifenóis/farmacologia , Traumatismos da Medula Espinal/tratamento farmacológico , Medula Espinal/metabolismo , Água/metabolismo , Alcenos/uso terapêutico , Animais , Citocinas/genética , Citocinas/metabolismo , Heme Oxigenase-1/genética , Heme Oxigenase-1/metabolismo , Locomoção , Masculino , NF-kappa B/genética , NF-kappa B/metabolismo , Fármacos Neuroprotetores/uso terapêutico , Ocludina/genética , Ocludina/metabolismo , Polifenóis/uso terapêutico , Ratos , Ratos Sprague-Dawley , Medula Espinal/efeitos dos fármacos , Traumatismos da Medula Espinal/metabolismo , Traumatismos da Medula Espinal/fisiopatologia , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo , Proteína da Zônula de Oclusão-1/genética , Proteína da Zônula de Oclusão-1/metabolismo
9.
Brain Res ; 1370: 220-6, 2011 Jan 25.
Artigo em Inglês | MEDLINE | ID: mdl-21092735

RESUMO

The study investigated the mechanism of the up-regulation of aquaporin-4 (AQP4) and aquaporin-1 (AQP1) expression induced by spinal cord injury (SCI). Using adult rat spinal cord injury model, it was found that up-regulation of hypoxia inducible factor-1α (HIF-1α), vascular endothelial growth factor (VEGF), AQP4, and AQP1 in response to spinal cord injury was greatly antagonized by 2-methoxyestradiol (2ME2), which can post-transcriptionally inhibit the expression of HIF-1α. VEGF alone significantly increased the extravasation of Evans blue and up-regulated the levels of AQP4 protein expression in the injured spinal cord issue, but the levels of AQP1 expression were not significantly changed. Taken together, our results suggest that expression of AQP4 and AQP1 is correlated with up-regulation of HIF-1α after SCI through the mechanisms that were dependent and independent of the VEGF signaling pathway, respectively. And the inhibitor of HIF-1α is a novel promising therapeutic agent for human SCI-induced edema in the future.


Assuntos
Aquaporina 1/antagonistas & inibidores , Aquaporina 4/antagonistas & inibidores , Estradiol/análogos & derivados , Fármacos Neuroprotetores/farmacologia , Traumatismos da Medula Espinal/tratamento farmacológico , 2-Metoxiestradiol , Animais , Aquaporina 1/metabolismo , Aquaporina 4/metabolismo , Modelos Animais de Doenças , Estradiol/farmacologia , Estradiol/uso terapêutico , Masculino , Fármacos Neuroprotetores/uso terapêutico , Ratos , Ratos Sprague-Dawley , Traumatismos da Medula Espinal/metabolismo , Moduladores de Tubulina/farmacologia , Moduladores de Tubulina/uso terapêutico , Regulação para Cima/efeitos dos fármacos , Regulação para Cima/fisiologia
10.
Brain Res ; 1357: 115-23, 2010 Oct 21.
Artigo em Inglês | MEDLINE | ID: mdl-20708606

RESUMO

The current study was performed to investigate the effect of baicalin (BC) on spinal cord injury (SCI) in rat. BC (10, 30 and 100mg/kg, i.p., respectively) was administered to rats immediately and every 24h following SCI. The BC therapy (100mg/kg) dramatically decreased (1) the water content of spinal cord tissue (by dry-wet weight method), (2) the permeability of blood-spinal cord barrier (measured by Evans blue), (3) oxidant stress (malondialdehyde values and glutathione levels evaluation), (4) proinflammatory cytokines expression (tumor necrosis factor-α and NF-κB) (5) and apoptosis (measured by Bax, Bcl-2 and Caspase-3 expression). And the treatment with BC also significantly improved the recovery of limb function (evaluated by motor recovery score). Taken together, our results clearly indicate that BC possesses potent anti-inflammatory and anti-apoptotic properties, attenuates the SCI and is a new promising therapeutic agent for human SCI in the future.


Assuntos
Barreira Hematoencefálica/efeitos dos fármacos , Barreira Hematoencefálica/metabolismo , Flavonoides/uso terapêutico , Compressão da Medula Espinal/tratamento farmacológico , Medula Espinal/efeitos dos fármacos , Análise de Variância , Animais , Apoptose/efeitos dos fármacos , Western Blotting , Água Corporal , Flavonoides/farmacologia , Glutationa/metabolismo , Imuno-Histoquímica , Masculino , Malondialdeído/metabolismo , Atividade Motora/fisiologia , NF-kappa B/metabolismo , Fármacos Neuroprotetores/farmacologia , Fármacos Neuroprotetores/uso terapêutico , Estresse Oxidativo/efeitos dos fármacos , Permeabilidade , Ratos , Ratos Sprague-Dawley , Recuperação de Função Fisiológica/efeitos dos fármacos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Medula Espinal/metabolismo , Medula Espinal/fisiopatologia , Compressão da Medula Espinal/metabolismo , Compressão da Medula Espinal/fisiopatologia , Fator de Necrose Tumoral alfa/metabolismo
11.
Zhonghua Nan Ke Xue ; 16(6): 552-5, 2010 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-20608363

RESUMO

More and more clinical evidence has confirmed the limitations of the use of serum PSA in the screening, detection and treatment of prostate cancer, and scientists are continuously seeking for new biomarkers of the disease. The discovery of early prostate cancer antigen 2 (EPCA-2) has provided a new base for the screening, detection, treatment and follow-up of prostate cancer.


Assuntos
Antígenos de Neoplasias/análise , Neoplasias da Próstata/diagnóstico , Diagnóstico Precoce , Humanos , Masculino
12.
Zhongguo Gu Shang ; 21(11): 836-8, 2008 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-19143246

RESUMO

OBJECTIVE: To observe the effects of neural stem cells (NSCs) transplantation on the brain derived neurotrophic factor (BDNF) after the spinal cord injury (SCI) of rats, and to investigate the mechanism of repairing the SCI by NSCs transplantation. METHODS: Neural stem cells were cultured from the hippocampus of rats' embryo and identified by immunocytochemistry. Seven days after the operation of SCI, the NSCs were transplanted into the injured site. Sixty adult Wistar rats were randomly divided into three groups: SCI cured with NSCs transplantation (group A), SCI received DMEM solution (group B), control group (group C). Then the expression of BDNF of the lesion and neighbor areas were examined by reverse transcsription polymerase chain reaction (RT-PCR) and immunohistochemistry, so as to investigated the mechanism of repairing the SCI after NSCS transplantation. RESULTS: According the RT-PCR results analysis, the expression of BDNF mRNA of group A enhanced higher than that of group B on the 1st, 3rd, 5th day after transplantation of NSCs. According the immunohistochemistry results analysis, the expression of BDNF mRNA of group A enhanced higher than that of group B on the 7th, 14th, 28th day similarly. CONCLUSION: The transplantation of NSCs can change the tiny-entironment by upregulating the expression of BDNF. It maybe one of the mechanism of repairing the SCI by NSCs transplantation.


Assuntos
Fator Neurotrófico Derivado do Encéfalo/genética , Transplante de Células-Tronco Mesenquimais , Neurônios/transplante , Traumatismos da Medula Espinal/terapia , Regulação para Cima , Animais , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Células Cultivadas , Modelos Animais de Doenças , Expressão Gênica , Humanos , Masculino , Neurônios/metabolismo , Distribuição Aleatória , Ratos , Ratos Wistar , Traumatismos da Medula Espinal/genética , Traumatismos da Medula Espinal/metabolismo , Traumatismos da Medula Espinal/cirurgia
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