Extracellular Ca(2+) influx and NO generation are inhibited by small interference RNA targeting extracellular Ca(2+)-sensing receptor in human umbilical vein endothelial cells / 生理学报
Acta Physiologica Sinica
;
(6): 289-295, 2012.
Artículo
en Chino
| WPRIM
| ID: wpr-335911
ABSTRACT
To investigate the effect of Ca(2+)-sensing receptor (CaR) on Spermine-induced extracellular Ca(2+) influx and NO generation in human umbilical vein endothelial cells (HUVEC), the small interference RNA (siRNA) specifically targeting CaR gene was designed, synthesized and transfected into HUVEC according to the cDNA sequence of human CaR gene in GenBank. The transfection efficiency and the interference efficiency of CaR protein were determined by laser scanning confocal microscopy and Western blot, respectively. Intracellular Ca(2+) concentration ([Ca(2+)](i)) was measured by Fura-2/AM loading. The production of NO and the activity of endothelial nitric oxide synthase (eNOS) were determined by the DAF-FM diacetate (DAF-FM DA). Western blot results demonstrated that siRNA targeting the CaR specifically decreased the expression of CaR protein in CaR siRNA group 48 h after transfection (P < 0.05). At the same time, the Spermine-induced [Ca(2+)](i), eNOS activity and NO generation were also significantly reduced (P < 0.05) in CaR siRNA group compared with those in the untransfected or negative siRNA transfected group. In conclusion, the present study suggests that the CaR plays an important role in the Spermine-evoked process of extracellular Ca(2+) influx and NO generation in HUVEC.
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Farmacología
/
Fisiología
/
Transfección
/
Espermina
/
Células Cultivadas
/
Calcio
/
ARN Interferente Pequeño
/
Receptores Sensibles al Calcio
/
Óxido Nítrico Sintasa de Tipo III
/
Células Endoteliales de la Vena Umbilical Humana
Límite:
Humanos
Idioma:
Chino
Revista:
Acta Physiologica Sinica
Año:
2012
Tipo del documento:
Artículo
Similares
MEDLINE
...
LILACS
LIS