De Mello Walmor C
School of Medicine, University of Puerto Rico , San Juan, PR , USA.
Front Endocrinol (Lausanne). 2015 Dec 11;6:185. doi: 10.3389/fendo.2015.00185. eCollection 2015.
The activation of the renin-angiotensin system is known to impair intercellular communication in the heart, but the role of aldosterone on the process of chemical communication and particularly the intercellular diffusion of glucose between cardiomyocytes is not known. This problem was investigated in cell pairs isolated from the left ventricle of adult Wistar Kyoto rats. For this, fluorescent glucose was dialyzed into one cell of the pair using the whole cell clamp technique, and its diffusion from cell-to-cell through gap junctions was followed by measuring the fluorescence intensity in the dialyzed as well as in non-dialyzed cell as a function of time. The results indicated that (1) in cell pairs exposed to aldosterone (100 nM) for 24 h, the intercellular flow of glucose through gap junctions was disrupted; (2) although the mechanism by which aldosterone disrupts the cell-to-cell flow of glucose is multifactorial, two major factors are involved: oxidative stress and PKC activation; (3) the effect of aldosterone was significantly reduced by spironolactone (100 nM); and (4) calculation of gap junction permeability (Pj) indicated an average values of 0.3 ± 0.001 × 10(-4) cm/s (n = 31) (four animals) for controls and 24 ± 0.03 × 10(-6) cm/s (n = 34) (four animals) (P < 0.05) for cell pairs exposed to aldosterone (100 nM) for 24 h. Bis-1 (10(-9)M), which is a selective PKC inhibitor, added to the aldosterone solution, improved the value of Pj to 0.21 ± 0.001 × 10(-4) cm/s (n = 24) (P < 0.05), whereas spironolactone (100 nM) added to aldosterone solution, reduced significantly the effect of the hormone on junctional permeability to glucose.
已知肾素-血管紧张素系统的激活会损害心脏中的细胞间通讯,但醛固酮在化学通讯过程中,特别是在心肌细胞之间葡萄糖的细胞间扩散中所起的作用尚不清楚。本研究在从成年Wistar Kyoto大鼠左心室分离的细胞对中对这一问题进行了探究。为此,使用全细胞钳技术将荧光葡萄糖透析到细胞对中的一个细胞中,并通过测量透析细胞和未透析细胞中荧光强度随时间的变化,来跟踪其通过缝隙连接在细胞间的扩散。结果表明:(1)在暴露于醛固酮(100 nM)24小时的细胞对中,通过缝隙连接的葡萄糖细胞间流动受到破坏;(2)尽管醛固酮破坏葡萄糖细胞间流动的机制是多因素的,但涉及两个主要因素:氧化应激和蛋白激酶C(PKC)激活;(3)螺内酯(100 nM)可显著降低醛固酮的作用;(4)缝隙连接通透性(Pj)的计算表明,对照组的平均值为0.3±0.001×10(-4) cm/s(n = 31)(四只动物),而暴露于醛固酮(100 nM)24小时的细胞对的平均值为24±0.03×10(-6) cm/s(n = 34)(四只动物)(P < 0.05)。添加到醛固酮溶液中的选择性PKC抑制剂双-1(10(-9)M)可将Pj值提高到0.21±0.001×10(-4) cm/s(n = 24)(P < 0.05),而添加到醛固酮溶液中的螺内酯(100 nM)则可显著降低该激素对葡萄糖连接通透性的影响。