黄芪水提取物促进小鼠胚胎干细胞向血管平滑肌细胞分化的实验研究

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/mL ARE, ARE 高剂量组（120 g/mL 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。

Experimental Study of Effect of Astragalus Radix Extract on Differentiation of Vascular Smooth Muscle Cell fromMouse Embryonic StemCells

Objective:To investigate the effects and possible mechanism of astragalus radix extract (ARE) on differentiation of vascular smooth muscle cells (VSMCs) from mouse embryonic stem cells (ESCs).Methods:VSMCs differentiation model was established by seeding ESCs directly on collagen IV coated plates. Cells were divided into 3 groups according to dose of ARE supplemented into culture medium: (1) control group (no ARE); (2) low dose group (60 g/mL); and (3) high dose group (120 g/mL). Cells were treated for 14 days. Immunofluorescence staining, real-time PCR and Western Blot were employed to detect expression of differentiation marker of VSMCs. The cholinergic receptor agonist (carbacol) was used to assess contractility of VSMCs from different groups. Real-time PCR was performed to detect changes of key factors regulating VSMCs differentiation.Results:Compared to the control group, ARE treatment significantly increased expression of VSMCs differentiation markers in a dose dependent manner. ARE treatment also improved contractility of VSMCs. In addition, ARE treatment promoted expression of transforming growth factor (TGF- ).Conclusion:ARE promotes VSMCs differentiation fromESCs possibly through activating TGF- pathway.