Department of Pharmacology, Center for Molecular Medicine, University of Nevada School of Medicine, Reno, Nevada.
Am J Physiol Cell Physiol. 2013 Sep 15;305(6):C632-42. doi: 10.1152/ajpcell.00324.2012. Epub 2013 Jun 26.
The mechanisms governing maintenance of quiescence during pregnancy remain largely unknown. The current study characterizes a stretch-activated, tetraethylammonium-insensitive K(+) current in smooth muscle cells isolated from pregnant human myometrium. This study hypothesizes that these K(+) currents can be attributed to TREK-1 and that upregulation of this channel during pregnancy assists with the maintenance of a negative cell membrane potential, conceivably contributing to uterine quiescence until full term. The results of this study demonstrate that, in pregnant human myometrial cells, outward currents at 80 mV increased from 4.8 ± 1.5 to 19.4 ± 7.5 pA/pF and from 3.0 ± 0.8 to 11.8 ± 2.7 pA/pF with application of arachidonic acid (AA) and NaHCO3, respectively, causing intracellular acidification. Similarly, outward currents were inhibited following application of 10 μM fluphenazine by 51.2 ± 9.8% after activation by AA and by 73.9 ± 4.2% after activation by NaHCO3. In human embryonic kidney (HEK-293) cells stably expressing TREK-1, outward currents at 80 mV increased from 91.0 ± 23.8 to 247.5 ± 73.3 pA/pF and from 34.8 ± 8.9 to 218.6 ± 45.0 pA/pF with application of AA and NaHCO3, respectively. Correspondingly, outward currents were inhibited 89.5 ± 2.3% by 10 μM fluphenazine following activation by AA and by 91.6 ± 3.4% following activation by NaHCO3. Moreover, currents in human myometrial cells were activated by stretch and were reduced by transfection with small interfering RNA or extracellular acidification. Understanding gestational regulation of expression and gating of TREK-1 channels could be important in determining appropriate maintenance of uterine quiescence during pregnancy.
维持妊娠期间静止状态的机制在很大程度上仍然未知。本研究描述了从妊娠人子宫平滑肌细胞中分离出的一种伸展激活的、四乙铵不敏感的 K(+)电流。本研究假设这些 K(+)电流可以归因于 TREK-1,并且在妊娠期间这种通道的上调有助于维持负细胞膜电位,这可能有助于子宫静止直到足月。本研究的结果表明,在妊娠人子宫平滑肌细胞中,80 mV 时的外向电流分别从 4.8 ± 1.5 增加到 19.4 ± 7.5 pA/pF 和从 3.0 ± 0.8 增加到 11.8 ± 2.7 pA/pF,用花生四烯酸 (AA) 和 NaHCO3 处理,导致细胞内酸化。同样,用 10 μM 氟奋乃静处理后,外向电流分别被 AA 激活抑制 51.2 ± 9.8%,被 NaHCO3 激活抑制 73.9 ± 4.2%。在稳定表达 TREK-1 的人胚肾 (HEK-293) 细胞中,80 mV 时的外向电流分别从 91.0 ± 23.8 增加到 247.5 ± 73.3 pA/pF 和从 34.8 ± 8.9 增加到 218.6 ± 45.0 pA/pF,用 AA 和 NaHCO3 处理。相应地,用 10 μM 氟奋乃静处理后,外向电流分别被 AA 激活抑制 89.5 ± 2.3%,被 NaHCO3 激活抑制 91.6 ± 3.4%。此外,人子宫平滑肌细胞的电流可被伸展激活,并可通过转染小干扰 RNA 或细胞外酸化来减少。了解妊娠期间 TREK-1 通道表达和门控的调节可能对确定妊娠期间子宫静止的适当维持很重要。