黄芪甲苷Ⅳ通过靶向Wnt/β-catenin信号通路调控低氧诱导的人肺动脉平滑肌细胞的增殖和迁移  

Astragaloside Ⅳ inhibited hypoxia-enhanced proliferation and migration of human pulmonary arterial smooth muscle cells through modulating Wnt/β-catenin signaling pathway

作　　者：赵梅[1] 邢诒雄[1] 陈鑫[2] ZHAO Mei;XING Yi-xiong;CHEN Xin(Department of Pharmacy,Sanya Central Hospital(Hainan Third People's Hospital),Sanya,Hainan 572000,China;Deparment of Neurosurgery,The First Affiliated Hospital of Harbin Medical University,Harbin,Heilongjiang 150001,China)

机构地区：[1]三亚中心医院(海南省第三人民医院)药学部,海南三亚572000 [2]哈尔滨医科大学附属第一医院神经外科,黑龙江哈尔滨150001

出　　处：《中国热带医学》2022年第8期762-768,共7页China Tropical Medicine

基　　金：海南省自然科学基金(No.822RC872);三亚市医疗卫生科技创新项目(No.2019YW09)。

摘　　要：目的探讨黄芪甲苷Ⅳ(astragalosideⅣ,AS-Ⅳ)对低氧诱导的人肺动脉平滑肌细胞增殖和迁移的影响及其相关分子机制。方法运用实时定量PCR和Western blot检测增殖细胞核抗原(PCNA)mRNA和蛋白的表达水平;CCK-8和EdU试验检测细胞活性和增殖;Transwell迁移试验检测细胞迁移。结果低氧处理人肺动脉平滑肌细胞24 h后,PCNA mRNA和蛋白表达量显著增加(P<0.05)。低氧刺激能显著增加人肺动脉平滑肌细胞的活性和增殖(P<0.05)。此外,低氧刺激还增强了人肺动脉平滑肌细胞的迁移能力(P<0.05)。AS-Ⅳ可下调低氧诱导的人肺动脉平滑肌细胞PNCA mRNA和蛋白的表达水平,抑制细胞活性、增殖以及迁移能力,其抑制作用呈浓度依赖性(P<0.05)。低氧诱导能够激活人肺动脉平滑肌细胞Wnt/β-catenin信号通路(P<0.05);而AS-Ⅳ能够浓度依赖性的抑制低氧诱导人肺动脉平滑肌细胞的Wnt/β-catenin信号通路的活性(P<0.05)。Wnt/β-catenin信号通路抑制剂XAV939可降低低氧诱导的人肺动脉平滑肌细胞PNCA mRNA和蛋白的的表达水平,同时降低低氧诱导的人肺动脉平滑肌细胞活性、增殖以及迁移能力(P<0.05)。Wnt/β-catenin信号转导激动剂LiCl处理可恢复AS-Ⅳ处理下调的人肺动脉平滑肌细胞PCNA mRNA和蛋白的表达量(P<0.05);AS-Ⅳ对低氧处理的人肺动脉平滑肌细胞增殖和迁移的抑制作用也被LiCl处理明显减弱(P<0.05)。结论研究表明AS-Ⅳ能够逆转低氧诱导的人肺动脉平滑肌细胞的增殖和迁移,此作用可能是通过调控Wnt/β-catenin信号通路实现的。ObjectiveThis study aims to determine the effects of astragalosideⅣ(AS-Ⅳ)treatment on the viability,proliferation and migration of hypoxia-stimulated human pulmonary arterial smooth muscle cells(PASMCs),and to explore the underlying molecular mechanisms.MethodsThe mRNA and protein expression levels of proliferating cell nuclear antigen(PCNA)were determined qRT-PCR and Western blot assays,respectively.Cell viability and cell proliferation were determined by CCK-8 and 5-ethynyl-2′-deoxyuridine(EdU)cell proliferation assays,respectively.The cell migration was determined by Transwell migration assay.ResultsHypoxia stimulation up-regulated the mRNA and protein expression of PCNA in PASMCs(P<0.05);hypoxia stimulation significantly promoted PASMC viability and proliferation(P<0.05),and also increased the migration of PASMCs(P<0.05).AS-Ⅳconcentration-dependently down-regulated the mRNA expression and protein expression of PCNA(P<0.05),inhibited the viability,proliferation and migration of PASMCs under hypoxia(P<0.05).Hypoxia stimulation activated the Wnt/β-catenin signaling in PASMCs;while AS-Ⅳconcentration-dependently repressed the Wnt/β-catenin signaling in the hypoxia-stimulated PASMCs(P<0.05).Moreover,the treatment of XAV393,a Wnt/β-catenin inhibitor,attenuated the hypoxia-induced increase in the viability,proliferation and migration of PASMCs(P<0.05).The treatment of LiCl,a Wnt/β-catenin activator,restored the mRNA and protein expression levels of PCNA in the hypoxia-stimulated PASMCs with AS-Ⅳtreatment(P<0.05).The inhibitory effects of AS-Ⅳtreatment on the viability,proliferation and migration of PASMCs under hypoxia was attenuated by LiCl treatment(P<0.05).ConclusionOur results indicate that AS-Ⅳreversed hypoxia-induced proliferation and migration of human pulmonary artery smooth muscle cells,which may be by regulating the Wnt/β-catenin signaling pathway.

关 键 词：肺动脉平滑肌细胞 黄芪甲苷Ⅳ 增殖 迁移 Wnt/β-catenin信号 

分 类 号：R966[医药卫生—药理学]