VEGF-A and VEGF-B Coordinate the Arteriogenesis to Repair the Infarcted Heart with Vagus Nerve Stimulation
VEGF-A and VEGF-B Coordinate the Arteriogenesis to Repair the Infarcted Heart with Vagus Nerve Stimulation
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VEGF-A 和 VEGF-B 协调动脉生成，通过迷走神经刺激修复梗塞心脏。
DOI：
10.1159/000491775
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发表时间：
2018-01-01
影响因子：
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通讯作者：
Tang, Jun-ming
中科院分区：
文献类型：
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作者：
Lv, Yan-xia;Zhong, Sen;Tang, Jun-ming
背景/目的：迷走神经刺激（VNS）可抑制心律失常活动，减少心肌细胞损伤。然而，VNS如何影响梗死心脏的血管生成/动脉生成尚不清楚。方法：采用冠状动脉左前降支结扎法致大鼠心肌梗死。LAD术后7天，在颈部左右迷走神经周围套上不锈钢丝进行迷走神经刺激（VNS）。以20 Hz、每分钟10秒、持续0.2ms的脉冲刺激迷走神经，连续4小时，ELISA法测定迷走神经刺激后心脏组织和血清中乙酰胆碱释放水平。VNS后3天和14天，分别采用Real-time PCR、免疫染色和western blot检测VNS心脏中VEGF-A/B的表达和α - sma -和cd31阳性血管，α - 7-nAChR阻滞剂甲胺（10 mg/kg, ip）或马赫- r阻滞剂阿托品（10 mg/kg, ip）预处理1小时。采用Langendorff系统和血流动力学参数分析VEGF-A/ b敲低或VEGFR阻滞剂AMG706预处理的vns心脏的冠状动脉功能和左心室功能。观察VNS刺激冠状动脉平滑肌细胞（VSMCs）后冠状动脉内皮细胞增殖、迁移和血管形成情况。结果：VNS刺激心肌梗死后冠状动脉平滑肌细胞（VSMCs）和内皮细胞（ECs）中VEGF-A和VEGF-B的表达，导致心肌梗死后α - sma和cd31阳性血管数量增加。在体内，m-AChR抑制剂阿托品和α - 7-nAChR阻滞剂甲胺可抑制vns诱导的VEGF-A/B表达和血管生成。有趣的是，shRNA敲低VEGF-A主要减少了vns介导的CD31（+）微血管的形成。相反，VEGF-B的抑制能有效地消除vns诱导的α - sma（+）血管的形成。与vns诱导的VEGF-B相比，vns诱导的VEGF-A对EC管形成的影响更大。VEGF-A促进体外培养的EC增殖和VSMC迁移，VEGF-B诱导体外培养的VSMC增殖和EC迁移。迷走神经递质乙酰胆碱在EC中通过m/nACh-R/PI3K/Akt/Sp1通路刺激VEGF-A/B的表达。功能上，VNS改善了冠状动脉功能和左心室功能。然而，拮抗剂AMG706阻断VEGF受体或shRNA敲低VEGF- a或VEGF- b可显著降低VNS对心室功能的有益作用。结论：VNS通过VEGF-A和VEGF-B的协同作用促进血管生成/动脉生成，修复心肌梗死。(C) 2018作者：s . Karger AG，巴塞尔出版
Background/Aims: Vagus nerve stimulation (VNS) suppresses arrhythmic activity and minimizes cardiomyocyte injury. However, how VNS affects angiogenesis/arteriogenesis in infarcted hearts, is poorly understood. Methods: Myocardial infarction (MI) was achieved by ligation of the left anterior descending coronary artery (LAD) in rats. 7 days after LAD, stainless-steel wires were looped around the left and right vagal nerve in the neck for vagus nerve stimulation (VNS). The vagal nerve was stimulated with regular pulses of 0.2ms duration at 20 Hz for 10 seconds every minute for 4 hours, and then ACh levels by ELISA in cardiac tissue and serum were evaluated for its release after VNS. Three and 14 days after VNS, Real-time PCR, immunostaining and western blot were respectively used to determine VEGF-A/B expressions and alpha-SMA- and CD31-postive vessels in VNS-hearts with pretreatment of alpha 7-nAChR blocker mecamylamine (10 mg/kg, ip) or mACh-R blocker atropine (10 mg/kg, ip) for 1 hour. The coronary function and left ventricular performance were analyzed by Langendorff system and hemodynamic parameters in VNS-hearts with pretreatment of VEGF-A/B-knockdown or VEGFR blocker AMG706. Coronary arterial endothelial cells proliferation, migration and tube formation were evaluated for angiogenesis following the stimulation of VNS in coronary arterial smooth muscle cells (VSMCs). Results: VNS has been shown to stimulate VEGF-A and VEGF-B expressions in coronary arterial smooth muscle cells (VSMCs) and endothelial cells (ECs) with an increase of alpha-SMA- and CD31-postive vessel number in infarcted hearts. The VNS-induced VEGF-A/B expressions and angiogenesis were abolished by m-AChR inhibitor atropine and alpha 7-nAChR blocker mecamylamine in vivo. Interestingly, knockdown of VEGF-A by shRNA mainly reduced VNS-mediated formation of CD31(+) microvessels. In contrast, knockdown of VEGF-B powerfully abrogated VNS-induced formation of alpha-SMA(+) vessels. Consistently, VNS-induced VEGF-A showed a greater effect on EC tube formation as compared to VNS-induced VEGF-B. Moreover, VEGF-A promoted EC proliferation and VSMC migration while VEGF-B induced VSMC proliferation and EC migration in vitro. Mechanistically, vagal neurotransmitter acetylcholine stimulated VEGF-A/B expressions through m/nACh-R/PI3K/Akt/Sp1 pathway in EC. Functionally, VNS improved the coronary function and left ventricular performance. However, blockade of VEGF receptor by antagonist AMG706 or knockdown of VEGF-A or VEGF-B by shRNA significantly diminished the beneficial effects of VNS on ventricular performance. Conclusion: VNS promoted angiogenesis/arteriogenesis to repair the infracted heart through the synergistic effects of VEGF-A and VEGF-B. (C) 2018 The Author(s) Published by S. Karger AG, Basel