Key Laboratory of Targeted Intervention of Cardiovascular Disease, Collaborative Innovation Center of Translational Medicine for Cardiovascular Disease, Department of Physiology, Nanjing Medical University, Nanjing, China.
Department of Basic Medicine, Wuxi School of Medicine, Jiangnan University, Wuxi, China.
Antioxid Redox Signal. 2019 Dec 20;31(18):1352-1370. doi: 10.1089/ars.2019.7723.
Vascular calcification (VC) is a hallmark feature of cardiovascular disease and a significant risk factor for morbidity and mortality. Salusin-β exerts cardiovascular regulating effects in hypertension, atherosclerosis, and diabetes. The present study was designed to examine the roles of salusin-β in the progression of VC and its downstream signaling mechanisms. Salusin-β expression in both the aortas of VC rats induced by vitamin D3 and nicotine and vascular smooth muscle cells (VSMCs) incubated with calcifying media was increased. Salusin-β knockdown remarkably reduced VC, whereas overexpression of salusin-β exacerbated VC both and . Overexpression of salusin-β promoted the VSMC osteochondrogenic transition, decreased Klotho protein levels, enhanced Ras-related C3 botulinum toxin substrate 1 (Rac1) activity and the translocation of p47phox to the membrane, increased the expression of nicotinamide adenine dinucleotide phosphate [NAD(P)H] oxidase subunits and the production of reactive oxygen species (ROS) with or without calcifying media; however, salusin-β deficiency played the opposite roles. The calcification and downregulated Klotho protein levels induced by salusin-β were restored by ROS scavenger N-acetyl-l-cysteine, diphenyleneiodonium chloride [an inhibitor of flavin-containing enzyme, including NAD(P)H oxidase], or gene knockdown of NAD(P)H oxidase (NOX)-2, p22phox, or p47phox but were not affected by NOX-1 and NOX-4 knockdown. Klotho knockdown attenuated the protective effect of salusin-β deficiency on VSMC calcification. By contrast, exogenous Klotho ameliorated the development of VC and ROS generation induced by salusin-β overexpression. Salusin-β is a critical modulator in VC. Salusin-β regulates VC through activation of NAD(P)H/ROS-mediated Klotho downregulation, suggesting that salusin-β may be a novel target for treatment of VC.
血管钙化 (VC) 是心血管疾病的一个显著特征，也是发病率和死亡率的重要危险因素。Salusin-β 在高血压、动脉粥样硬化和糖尿病中发挥心血管调节作用。本研究旨在探讨 salusin-β 在 VC 进展中的作用及其下游信号机制。维生素 D3 和尼古丁诱导的 VC 大鼠主动脉和钙化培养基孵育的血管平滑肌细胞 (VSMC) 中 salusin-β 的表达增加。Salusin-β 敲低显著减少 VC，而过表达 salusin-β 则加剧 VC。过表达 salusin-β 促进 VSMC 成骨软骨转化，降低 Klotho 蛋白水平，增强 Ras 相关 C3 肉毒杆菌毒素底物 1 (Rac1) 活性和 p47phox 向膜的易位，增加烟酰胺腺嘌呤二核苷酸磷酸 [NAD(P)H] 氧化酶亚基的表达和活性氧 (ROS) 的产生，无论是否存在钙化培养基；然而，salusin-β 缺乏则起到相反的作用。ROS 清除剂 N-乙酰-l-半胱氨酸、二苯碘 [一种包含黄素酶的抑制剂，包括 NAD(P)H 氧化酶] 或 NAD(P)H 氧化酶 (NOX)-2、p22phox 或 p47phox 的基因敲低可恢复由 salusin-β 诱导的钙化和下调的 Klotho 蛋白水平，但不影响 NOX-1 和 NOX-4 的敲低。Klotho 敲低减弱了 salusin-β 缺乏对 VSMC 钙化的保护作用。相比之下，外源性 Klotho 改善了由 salusin-β 过表达引起的 VC 发展和 ROS 生成。Salusin-β 是 VC 的关键调节剂。Salusin-β 通过激活 NAD(P)H/ROS 介导的 Klotho 下调来调节 VC，表明 salusin-β 可能是治疗 VC 的新靶点。
