Li Shan-Liang, Yan Jie, Zhang Yan-Qiu, Zhen Chang-Lin, Liu Ming-Yu, Jin Jing, Gao Jin-Lai, Xiao Xiao-Lin, Shen Xin, Tai Yu, Hu Nan, Zhang Xin-Zi, Sun Zhi-Jie, Dong De-Li
Department of Pharmacology (The State-Province Key Laboratories of Biomedicine-Pharmaceutics of China, Key Laboratory of Cardiovascular Research, Ministry of Education), College of Pharmacy, Harbin Medical University, Translational Medicine Research and Cooperation Center of Northern China, Heilongjiang Academy of Medical Sciences, Harbin Medical University, PR China.
Institute of Materials Processing and Intelligent Manufacturing & Center for Biomedical Materials and Engineering, Harbin Engineering University, PR China.
Pharmacol Res. 2017 Jan;115:78-86. doi: 10.1016/j.phrs.2016.11.008. Epub 2016 Nov 18.
We previously demonstrated that the typical mitochondrial uncoupler carbonyl cyanide m-chlorophenylhydrazone (CCCP) inhibited artery constriction, but CCCP was used only as a pharmacological tool. Niclosamide is an anthelmintic drug approved by FDA. Niclosamide ethanolamine (NEN) is a salt form of niclosamide and has been demonstrated to uncouple mitochondrial oxidative phosphorylation. The aim of the present study was to elucidate the vasoactivity of NEN and the potential mechanisms. Isometric tension of rat mesenteric artery and thoracic aorta was recorded by using multi-wire myograph system. The protein levels were measured by using western blot techniques. Niclosamide ethanolamine (NEN) treatment relaxed phenylephrine (PE)- and high K (KPSS)-induced constriction, and pre-treatment with NEN inhibited PE- and KPSS-induced constriction of rat mesenteric arteries. In rat thoracic aorta, NEN also showed antagonism against PE- and KPSS-induced constriction. NEN induced increase of cellular ADP/ATP ratio in vascular smooth muscle cells (A10) and activated AMP-activated protein kinase (AMPK) in A10 cells and rat thoracic aorta. NEN-induced aorta relaxation was attenuated in AMPKα1 knockout (-/-) mice. SERCA inhibitors cyclopiazonic acid and thapsigargin, but not K channel blockers glibenclamide and 5-hydroxydecanoic acid, attenuated NEN-induced vasorelaxation in rat mesenteric arteries. NEN treatment increased cytosolic [Ca] and depolarized mitochondrial membrane potential in vascular smooth muscle cells (A10). Niclosamide in non-salt form showed the similar vasoactivity as NEN in rat mesenteric arteries. Niclosamide ethanolamine inhibits artery constriction, indicating that it would be promising to be developed as an anti-hypertensive drug or it would induce vasodilation-related side effects when absorbed in vivo.
我们之前证明，典型的线粒体解偶联剂羰基氰化物间氯苯腙（CCCP）可抑制动脉收缩，但CCCP仅用作一种药理学工具。氯硝柳胺是一种经美国食品药品监督管理局（FDA）批准的驱虫药。氯硝柳胺乙醇胺（NEN）是氯硝柳胺的盐形式，已被证明可使线粒体氧化磷酸化解偶联。本研究的目的是阐明NEN的血管活性及其潜在机制。使用多线肌张力描记系统记录大鼠肠系膜动脉和胸主动脉的等长张力。采用蛋白质印迹技术测量蛋白质水平。氯硝柳胺乙醇胺（NEN）处理可舒张去甲肾上腺素（PE）和高钾（KPSS）诱导的收缩，并且用NEN预处理可抑制PE和KPSS诱导的大鼠肠系膜动脉收缩。在大鼠胸主动脉中，NEN也显示出对PE和KPSS诱导的收缩的拮抗作用。NEN诱导血管平滑肌细胞（A10）中细胞ADP/ATP比值升高，并激活A10细胞和大鼠胸主动脉中的AMP激活蛋白激酶（AMPK）。在AMPKα1基因敲除（-/-）小鼠中，NEN诱导的主动脉舒张减弱。肌浆网Ca2+-ATP酶抑制剂环匹阿尼酸和毒胡萝卜素可减弱NEN诱导的大鼠肠系膜动脉舒张，但钾通道阻滞剂格列本脲和5-羟基癸酸则无此作用。NEN处理可增加血管平滑肌细胞（A10）中的胞质[Ca2+]并使线粒体膜电位去极化。非盐形式的氯硝柳胺在大鼠肠系膜动脉中显示出与NEN相似的血管活性。氯硝柳胺乙醇胺可抑制动脉收缩，表明其有潜力被开发为抗高血压药物，或者在体内吸收时会诱发与血管舒张相关的副作用。
