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Identification of novel DNA methylation inhibitors via a two-component reporter gene system |
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| Authors: | Yi-Shiuan Lin Arthur Y Shaw Shi-Gang Wang Chia-Chen Hsu I-Wen Teng Min-Jen Tseng Tim HM Huang Ching-Shih Chen Yu-Wei Leu Shu-Huei Hsiao |
| Institution: | 1. Department of Pathophysiology, Qiqihar Medical University, 161006, Qiqihar, PR China 2. Department of Pathophysiology, Harbin Medical University, 150086, Harbin, PR China 5. The second affiliated hospital of Harbin Medical University, 150086, Harbin, PR China 4. Bio-pharmaceutical Key Laboratory of Heilongjiang Province, 150086, Harbin, PR China 3. Department of Pharmacology, Harbin Medical University, 150086, Harbin, PR China 6. Department of Biology, Lakehead University, P7B5E1, Thunder Bay, Ont, Canada |
| Abstract: | BackgroundThe extracellular calcium-sensing receptor (CaSR) belongs to family C of the G protein coupled receptors. Whether the CaSR is expressed in the pulmonary artery (PA) is unknown.MethodsThe expression and distribution of CaSR were detected by RT-PCR, Western blotting and immunofluorescence. PA tension was detected by the pulmonary arterial ring technique, and the intracellular calcium concentration (Ca2+]i) was detected by a laser-scanning confocal microscope.ResultsThe expressions of CaSR mRNA and protein were found in both rat pulmonary artery smooth muscle cells (PASMCs) and PAs. Increased levels of Ca2+]o (extracellular calcium concentration) or Gd3+ (an agonist of CaSR) induced an increase of Ca2+]i and PAs constriction in a concentration-dependent manner. In addition, the above-mentioned effects of Ca2+ and Gd3+ were inhibited by U73122 (specific inhibitor of PLC), 2-APB (specific antagonist of IP3 receptor), and thapsigargin (blocker of sarcoplasmic reticulum calcium ATPase).ConclusionsCaSR is expressed in rat PASMCs, and is involved in regulation of PA tension by increasing Ca2+]i through G-PLC-IP3 pathway. |
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