Liu Daoyan, Scholze Alexandra, Zhu Zhiming, Kreutz Reinhold, Wehland-von-Trebra Markus, Zidek Walter, Tepel Martin
Medizinische Klinik IV, Charité Campus Benjamin Franklin, Berlin, Germany.
Am J Hypertens. 2005 Nov;18(11):1503-7. doi: 10.1016/j.amjhyper.2005.05.033.
Disturbances in the regulation of cytosolic calcium concentration have been attributed to primary hypertension, but the role of calcium-permeable transient receptor potential canonical channel 3 (TRPC3) has not yet been evaluated in primary hypertension.
Expression of TRPC3 was determined using in-cell Western assay. Evaluation of RNA interference for the downregulation of a specific gene in cells by small interfering RNA was performed. Measurements of cytosolic calcium were carried out using the fluorescent dye fura2.
Expression of TRPC3 was significantly increased in monocytes from spontaneously hypertensive rats (SHR) compared with normotensive Wistar-Kyoto rats (WKY). Transplasmamembrane calcium influx and thapsigargin-induced sustained calcium increase were significantly higher in SHR compared with WKY. In the presence of the TRP channel blocker SKF-96365 these differences were no longer observed. Specific TRPC3-knockdown by transfection of monocytes from SHR with small interfering RNA significantly reduced TRPC3 expression, trans-plasma membrane calcium influx, and thapsigargin-induced sustained calcium increase.
This study shows, for the first time, increased TRPC3 channel expression and increased TRPC3-related calcium influx in SHR.
细胞溶质钙浓度调节紊乱被认为与原发性高血压有关，但钙通透性瞬时受体电位香草酸受体3（TRPC3）在原发性高血压中的作用尚未得到评估。
采用细胞内蛋白质免疫印迹法测定TRPC3的表达。通过小干扰RNA对细胞中特定基因下调进行RNA干扰评估。使用荧光染料fura2进行细胞溶质钙的测量。
与正常血压的Wistar-Kyoto大鼠（WKY）相比，自发性高血压大鼠（SHR）单核细胞中TRPC3的表达显著增加。与WKY相比，SHR的跨膜钙内流和毒胡萝卜素诱导的持续性钙增加显著更高。在存在TRP通道阻滞剂SKF-96365的情况下，这些差异不再观察到。用小干扰RNA转染SHR的单核细胞进行特异性TRPC3基因敲低，可显著降低TRPC3表达、跨质膜钙内流和毒胡萝卜素诱导的持续性钙增加。
本研究首次表明，SHR中TRPC3通道表达增加以及与TRPC3相关的钙内流增加。
