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目的:研究黄芪水提取物(Astragalus radix extract,ARE)对小鼠胚胎干细胞(Embryonic stem cells,ESCs)来源的血管平滑肌细胞(Vascular smooth muscle cells,VSMCs)分化的影响。方法:将ESCs直接接种到IV型胶原包被的培养皿中建立VSMC分化模型,向培养液中添加不同剂量ARE(对照组不添加ARE,ARE低剂量组60 g/m L ARE,ARE高剂量组(120 g/m L ARE),将3组细胞连续培养14天。分别应用免疫荧光染色、实时定量PCR及Western Blot技术检测各组VSMC分化标志物表达情况。应用胆碱能受体激动剂卡巴可刺激各组细胞检测收缩反应。应用实时定量PCR技术检测调控VSMC分化的重要因子的变化。结果:与对照组相比,ARE处理后VSMC分化标志物表达显著增加,且呈剂量依赖性。ARE处理可提高VSMCs的收缩能力。此外,ARE处理后,转化生长因子(Transforming growth factor)的表达显著增加。结论:黄芪水提取物可能通过激活TGF-促进ESCs向VS MCs分化。
Objective: To investigate the effect of astragalus radix extract (ARE) on the differentiation of mouse vascular smooth muscle cells (VSMCs) derived from embryonic stem cells (ESCs). METHODS: VSCs were directly inoculated into IV collagen-coated culture dishes to establish VSMC differentiation models. Different doses of ARE were added to the medium (ARE, ARE low-dose group 60 g / mL ARE, ARE high dose (120 g / m L ARE), and the three groups of cells were cultured continuously for 14 days.Immunofluorescence staining, real-time quantitative PCR and Western Blot were used to detect the expression of VSMC differentiation markers in each group.Using cholinergic receptor agonist Carbamate stimulated the contractile response in all groups of cells, and the changes of important factors regulating the differentiation of VSMCs were detected by real-time quantitative PCR.Results: Compared with the control group, the expression of VSMCs increased significantly and in a dose-dependent manner. ARE treatment could increase VSMCs contractility.In addition, the expression of Transforming growth factor was significantly increased after treatment with ARE.Conclusion: Astragalus membranaceus water extract may promote the differentiation of ESCs into VS MCs by activating TGF-β1.