From the Departamento de Bioquímica y Biología Molecular y Fisiología, Universidad de Valladolid, Spain (M.A.-M., P.C., N.G.-M., S.M.-E., J.S., J.R.L.-L., M.T.P.-G.).
Instituto de Biología y Genética Molecular (IBGM), CSIC, Universidad de Valladolid, Spain (M.A.-M., P.C., N.G.-M., S.M.-E., J.S., M.S., M.A.d.l.F.).
Arterioscler Thromb Vasc Biol. 2019 Dec;39(12):e273-e286. doi: 10.1161/ATVBAHA.119.313492. Epub 2019 Oct 10.
We have previously described that changes in the expression of Kv channels associate to phenotypic modulation (PM), so that Kv1.3/Kv1.5 ratio is a landmark of vascular smooth muscle cells phenotype. Moreover, we demonstrated that the Kv1.3 functional expression is relevant for PM in several types of vascular lesions. Here, we explore the efficacy of Kv1.3 inhibition for the prevention of remodeling in human vessels, and the mechanisms linking the switch in Kv1.3 /Kv1.5 ratio to PM. Approach and Results: Vascular remodeling was explored using organ culture and primary cultures of vascular smooth muscle cells obtained from human vessels. We studied the effects of Kv1.3 inhibition on serum-induced remodeling, as well as the impact of viral vector-mediated overexpression of Kv channels or myocardin knock-down. Kv1.3 blockade prevented remodeling by inhibiting proliferation, migration, and extracellular matrix secretion. PM activated Kv1.3 via downregulation of Kv1.5. Hence, both Kv1.3 blockers and Kv1.5 overexpression inhibited remodeling in a nonadditive fashion. Finally, myocardin knock-down induced vessel remodeling and Kv1.5 downregulation and myocardin overexpression increased Kv1.5, while Kv1.5 overexpression inhibited PM without changing myocardin expression.
We demonstrate that Kv1.5 channel gene is a myocardin-regulated, vascular smooth muscle cells contractile marker. Kv1.5 downregulation upon PM leaves Kv1.3 as the dominant Kv1 channel expressed in dedifferentiated cells. We demonstrated that the inhibition of Kv1.3 channel function with selective blockers or by preventing Kv1.5 downregulation can represent an effective, novel strategy for the prevention of intimal hyperplasia and restenosis of the human vessels used for coronary angioplasty procedures.
我们之前曾描述过 Kv 通道表达的变化与表型调节（PM）有关，因此 Kv1.3/Kv1.5 比值是血管平滑肌细胞表型的标志。此外，我们证明 Kv1.3 的功能表达与几种类型的血管病变中的 PM 有关。在这里，我们探索了 Kv1.3 抑制在预防人血管重塑中的功效，以及 Kv1.3/Kv1.5 比值变化与 PM 相关的机制。
使用器官培养和从人血管获得的血管平滑肌细胞原代培养来探索血管重塑。我们研究了 Kv1.3 抑制对血清诱导的重塑的影响，以及病毒载体介导的 Kv 通道过表达或心肌抑制素敲低的影响。Kv1.3 阻断通过抑制增殖、迁移和细胞外基质分泌来预防重塑。PM 通过下调 Kv1.5 激活 Kv1.3。因此，Kv1.3 阻断剂和 Kv1.5 过表达以非加性方式抑制重塑。最后，心肌抑制素敲低诱导血管重塑和 Kv1.5 下调，心肌抑制素过表达增加 Kv1.5，而 Kv1.5 过表达抑制 PM 而不改变心肌抑制素表达。
我们证明 Kv1.5 通道基因是一种受心肌抑制素调节的血管平滑肌细胞收缩标志物。PM 时 Kv1.5 的下调使 Kv1.3 成为去分化细胞中表达的主要 Kv1 通道。我们证明，用选择性阻断剂抑制 Kv1.3 通道功能或防止 Kv1.5 下调可代表预防用于冠状动脉成形术的人血管内膜增生和再狭窄的有效、新颖策略。
