Increased Transient Receptor Potential Canonical Type 3 Channels in Vasculature From Hypertensive Rats
Increased Transient Receptor Potential Canonical Type 3 Channels in Vasculature From Hypertensive Rats
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DOI：
10.1161/hypertensionaha.108.116947
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发表时间：
2009-01-01
期刊：
影响因子：
8.3
通讯作者：
Tepel, Martin
中科院分区：
文献类型：
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作者：
Liu, Daoyan;Yang, Dachun;Tepel, Martin
我们测试的假设，瞬时受体电位典型3型（TRPC 3）通道增加血管平滑肌细胞和主动脉组织自发性高血压大鼠（SHR）与血压正常的Wistar京都大鼠相比。免疫组化和Western blotting检测TRPC 3的表达。使用pAdEasy-1系统通过特异性小干扰RNA和TRPC 3过表达进行TRPC 3基因敲除。用荧光分光光度法测定胞浆钙，用力传感器测定主动脉环的血管收缩。与Wistar京都大鼠相比，SHR大鼠主动脉环（1.48 +/- 0.05对1.00 +/- 0.06;各n = 6; P < 0.01）和血管平滑肌细胞（1.28 +/- 0.08对1.00 +/- 0.03;各n = 6; P < 0.05）中TRPC 3通道蛋白的表达显著更高。通过特异性小干扰RNA敲低TRPC 3基因表达可显著降低血管紧张素II诱导的钙内流30 +/- 3%（n = 6; P < 0.01），而TRPC 3过表达可显著增加钙内流55 +/- 3%（n = 6; P < 0.01）。与Wistar京都大鼠相比，SHR血管平滑肌细胞中血管紧张素II诱导的钙增加显著增强，即使在钙通道阻滞剂的存在下。血管紧张素II使SHR血管平滑肌细胞TRPC 3通道蛋白表达从1.28 ± 0.08显著升高至1.61 ± 0.08（各n = 6; P < 0.01）。血管紧张素II诱导的TRPC 3表达被替米沙坦阻止。给予SHR替米沙坦4周可显著降低血压、血管紧张素II诱导的血管收缩和主动脉组织中TRPC 3通道蛋白表达。TRPC 3的表达在SHR中使用降血压后没有显著降低。总之，我们给出的实验证据表明，增加TRPC 3通道蛋白在血管系统中的表达是重要的血压升高。（高血压。2009; 53：70-76.）
We tested the hypothesis that transient receptor potential canonical type 3 (TRPC3) channels are increased in vascular smooth muscle cells and aortic tissue from spontaneously hypertensive rats (SHR) compared with normotensive Wistar Kyoto rats. Expression of TRPC3 was analyzed by immunohistochemistry and Western blotting. TRPC3 gene knockdown was performed by specific small interfering RNA and TRPC3 overexpression using the pAdEasy-1 system. Cytosolic calcium was measured using fluorescence spectrophotometry and vasoconstriction of aortic rings using a force transducer. In SHR, the expression of TRPC3 channel protein was significantly higher in aortic rings (1.48 +/- 0.05 versus 1.00 +/- 0.06; each n = 6; P < 0.01) and vascular smooth muscle cells (1.28 +/- 0.08 versus 1.00 +/- 0.03; each n = 6; P < 0.05) compared with Wistar Kyoto rats. Knockdown of TRPC3 gene expression by specific small interfering RNA significantly reduced the angiotensin II-induced calcium influx by 30 +/- 3% (n = 6; P < 0.01), whereas TRPC3 overexpression significantly increased it by 55 +/- 3% (n = 6; P < 0.01). The angiotensin II-induced calcium increase was significantly enhanced in vascular smooth muscle cells from SHR compared with Wistar Kyoto rats, even in the presence of the calcium channel blocker amlodipine. Angiotensin II significantly elevated the TRPC3 channel protein expression in vascular smooth muscle cells from SHR from 1.28 +/- 0.08 to 1.61 +/- 0.08 (each n = 6; P < 0.01). Angiotensin II-induced TRPC3 expression was prevented by telmisartan. Administration of telmisartan to SHR for 4 weeks significantly reduced blood pressure, angiotensin II-induced vasoconstriction, and TRPC3 channel protein expression in aortic tissue. TRPC3 expression was not significantly reduced after reduction of blood pressure in SHR using amlodipine. In conclusion, we give experimental evidence that increased TRPC3 channel protein expression in the vasculature is important for elevated blood pressure. (Hypertension. 2009; 53: 70-76.)