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1.
Neuroscience Bulletin ; (6): 215-220, 2007.
Artículo en Inglés | WPRIM | ID: wpr-300961

RESUMEN

<p><b>OBJECTIVE</b>To investigate the regulatory effects of nerve growth factor (NGF) on basal and capsaicin-induced release of neuropeptide substance P (SP) in primary cultured embryonic rat dorsal root ganglion (DRG) neurons.</p><p><b>METHODS</b>DRGs were dissected from 15-day-old embryonic Wistar rats. DRG neurons were dissociated and cultured, and then exposed to different concentrations of NGF (10 ng/mL, 30 ng/mL, or 100 ng/mL) for 72 h. The neurons cultured in media without NGF served as control. RT-PCR were used for detecting the mRNAs of SP and vanilloid receptor 1 (VR1) in the DRG neurons. The SP basal and capsaicin (100 nmol/L)-induced release in the culture were measured by radioimmunoassay (RIA).</p><p><b>RESULTS</b>SP mRNA and VR1 mRNA expression increased in primary cultured DRG neurons in a dose-dependent manner of NGF. Both basal release and capsaicin-evoked release of SP increased in NGF-treated DRG neurons compared with in control group. The capsaicin-evoked release of SP also increased in a dose-dependent manner of NGF.</p><p><b>CONCLUSION</b>NGF may promote both basal release and capsaicin-evoked release of SP. NGF might increase the sensitivity of nociceptors by increasing the SP mRNA or VR1 mRNA.</p>


Asunto(s)
Animales , Ratas , Analgésicos no Narcóticos , Farmacología , Capsaicina , Farmacología , Células Cultivadas , Relación Dosis-Respuesta a Droga , Embrión de Mamíferos , Ganglios Espinales , Biología Celular , Regulación de la Expresión Génica , Factor de Crecimiento Nervioso , Farmacología , Neuronas , ARN Mensajero , Metabolismo , Radioinmunoensayo , Métodos , Ratas Wistar , Sustancia P , Genética , Metabolismo
2.
Chinese Journal of Hepatology ; (12): 291-294, 2003.
Artículo en Chino | WPRIM | ID: wpr-344418

RESUMEN

<p><b>OBJECTIVE</b>To study the specific expression of the antisense RNA against hepatitis B virus X (HBX) gene in hepatoblastoma cell line and its anti -HBV activity.</p><p><b>METHODS</b>HBX gene (nt.1370-1827) was amplified by PCR, then cloned into EB virus vector pEBAF which contained human alpha-fetoprotein promoter and enhancer. After transfected into 2.2.15 hepatoma cells and ECV304 human endothelial cells by lipofectin, northern blot, ELISA and real-time qualitative PCR were carried out to assay the expression of HBX mRNA, HBV antigens and HBV DNA level, respectively.</p><p><b>RESULTS</b>The HBX antisense RNA expression vector pEBAF-as-HBX which could be expressed specifically in 2.2.15 hepatoblastoma cells was successfully constructed. Both HBV DNA level and the expressions of hepatitis B virus surface antigen (HBsAg) and e antigen (HBeAg) in 2.2.15 hepatoblastoma cells were inhibited by pEBAF-as-HBX. Compared with those in sense control (pEBAF-s-HBX), the inhibitory rates of HBsAg, HBeAg, and HBV DNA were 37.9%, 36.8%, and 25%, respectively.</p><p><b>CONCLUSIONS</b>The pEBAF-as-HBX expression vector may lead to targeted-expression of HBX antisense RNA in hepatoma cells and shows great inhibition effect on HBV.</p>


Asunto(s)
Animales , Humanos , Carcinoma Hepatocelular , Genética , Patología , Virología , Línea Celular Tumoral , Replicación del ADN , Elementos de Facilitación Genéticos , Genética , Regulación Viral de la Expresión Génica , Terapia Genética , Métodos , Virus de la Hepatitis B , Genética , Fisiología , Neoplasias Hepáticas , Genética , Patología , Virología , Regiones Promotoras Genéticas , Genética , ARN sin Sentido , Farmacología , Transactivadores , Genética , Activación Transcripcional , Transfección , alfa-Fetoproteínas , Genética
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