Issue Archive

[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.] [Article in Chinese]

LIANG Xiao, LUO Xiao-Lin, ZHONG Hua, HU Qing-Hua, HE Fang^*

Department of Pathophysiology/Key Laboratory of Education Ministry of Xinjiang Endemic and Ethnic Diseases, Medical College of Shihezi University, Shihezi 832002, China； Department of Pathophysiology, Tongji Medical College, Huazhong University of Science and Technology/Key Laboratory for Respiratory Diseases, Health Ministry of China, Wuhan 430030, China

Abstract

To investigate the effect of Ca2+-sensing receptor (CaR) on Spermine-induced extracellular Ca2+ 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 Ca2+ concentration ([Ca2+]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 [Ca2+]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 Ca2+ influx and NO generation in HUVEC.

Key words: Endothelial cells; calcium-sensing receptors; calcium signaling; nitric oxide; small interference RNA

Received: 2011-12-03　　Accepted: 2012-02-17

Corresponding author: 何芳　　E-mail: fangf2002shz@126.com

Citing This Article：

LIANG Xiao, LUO Xiao-Lin, ZHONG Hua, HU Qing-Hua, HE Fang. [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.] [Article in Chinese]. Acta Physiol Sin 2012; 64 (3): 289-295 (in Chinese with English abstract).