Li Wen-Qun, Li Xiao-Hui, Du Jie, Zhang Wang, Li Dai, Xiong Xiao-Ming, Li Yuan-Jian
Department of Pharmacology, School of Pharmaceutical Sciences, Central South University, Changsha, 410078, China.
National Institution of Drug Clinical Trial, Xiangya Hospital, Central South University, Changsha, 410078, China.
Naunyn Schmiedebergs Arch Pharmacol. 2016 Jul;389(7):757-67. doi: 10.1007/s00210-016-1240-8. Epub 2016 Apr 6.
Rutaecarpine has been shown to exhibit wide pharmacological effects in the cardiovascular system via stimulation of calcitonin gene-related peptide (CGRP) release. In the present study, the effect of rutaecarpine on hypoxia-induced right ventricular (RV) remodeling and the underlying mechanisms were evaluated. RV remodeling was induced by hypoxia (10 % O2, 3 weeks) in rats. Rats were treated with rutaecarpine (20 or 40 mg/kg) by intragastric administration. Proliferation of cardiac fibroblasts was induced by TGF-β1 (5 ng/mL) and determined by MTS and EdU incorporation method. Cardiac fibroblasts were treated with exogenous CGRP (10 or 100 nM). The concentrations of CGRP and TGF-β1 in plasma were measured by ELISA. The expression of eIF3a, p27, α-SMA, collagen-I/III, ANP, and BNP were measured by real-time PCR or western blot. Hypoxia induced an increase of right ventricle systolic pressure (RVSP), ration of RV/LV+S, and RV/tibial length in rats, while cardiac hypertrophy, apoptosis, and fibrosis were detected. The expression of ANP, BNP, α-SMA, collagen-I, collagen-III, eIF3a, and TGF-β1 was up-regulated, and the expression of p27 was down-regulated in the right ventricle of hypoxia-treated rats. The plasma concentration of CGRP was decreased and TGF-β1 was increased in hypoxia-treated rats. All of these effects induced by hypoxia were attenuated by rutaecarpine in a dose-dependent manner. In cultured cardiac fibroblasts, TGF-β1 significantly promoted the proliferation and up-regulated the expression of α-SMA and collagen-I/III, while the expression of eIF3a was up-regulated and the expression of p27 was down-regulated. The effects of TGF-β1 were attenuated by CGRP. CGRP8-37, a selective CGRP receptor antagonist, abolished the effects of CGRP. Rutaecarpine attenuates hypoxia-induced RV remodeling via stimulation of CGRP release, and the effects of rutaecarpine involve the eIF3a/p27 pathway.
吴茱萸次碱已被证明可通过刺激降钙素基因相关肽（CGRP）释放，在心血管系统中发挥广泛的药理作用。在本研究中，评估了吴茱萸次碱对缺氧诱导的右心室（RV）重塑的影响及其潜在机制。通过在大鼠中制造缺氧环境（10%氧气，3周）来诱导RV重塑。通过灌胃给予大鼠吴茱萸次碱（20或40mg/kg）。用转化生长因子-β1（TGF-β1，5ng/mL）诱导心脏成纤维细胞增殖，并通过MTS和EdU掺入法进行测定。用外源性CGRP（10或100nM）处理心脏成纤维细胞。通过酶联免疫吸附测定法（ELISA）测量血浆中CGRP和TGF-β1的浓度。通过实时聚合酶链反应（PCR）或蛋白质免疫印迹法（western blot）测量真核翻译起始因子3a（eIF3a）、p27、α-平滑肌肌动蛋白（α-SMA）、I/III型胶原蛋白、心钠素（ANP）和脑钠肽（BNP）的表达。缺氧导致大鼠右心室收缩压（RVSP）、RV/LV+S比值和RV/胫骨长度增加，同时检测到心脏肥大、细胞凋亡和纤维化。在缺氧处理大鼠的右心室中，ANP、BNP、α-SMA、I型胶原蛋白、III型胶原蛋白、eIF3a和TGF-β1的表达上调，而p27的表达下调。缺氧处理大鼠的血浆CGRP浓度降低，TGF-β1浓度升高。吴茱萸次碱以剂量依赖的方式减弱了缺氧诱导的所有这些效应。在培养的心脏成纤维细胞中，TGF-β1显著促进增殖并上调α-SMA和I/III型胶原蛋白的表达，而eIF3a的表达上调，p27的表达下调。CGRP减弱了TGF-β1的作用。CGRP8-37，一种选择性CGRP受体拮抗剂，消除了CGRP的作用。吴茱萸次碱通过刺激CGRP释放减弱缺氧诱导的RV重塑，且吴茱萸次碱的作用涉及eIF3a/p27信号通路。
