基础医学与临床 ›› 2015, Vol. 35 ›› Issue (1): 26-32.

• 研究论文 • 上一篇    下一篇

ox-LDL抑制大鼠MSCs向内皮分化

吕铁伟1,孙慧超2,刘玲娟2,张蕾3,郑敏2,朱静2,刘振国4,田杰5   

  1. 1. 重庆医科大学附属儿童医院心血管内科
    2. 重庆医科大学附属儿童医院
    3. 重庆医科大学儿童医院
    4. 美国俄亥俄州立大学医学中心
    5. 重庆医科大学及附属儿童医院
  • 收稿日期:2013-11-18 修回日期:2014-04-18 出版日期:2015-01-05 发布日期:2014-12-30
  • 通讯作者: 吕铁伟 E-mail:ltw200145@163.com
  • 基金资助:
    氧化应激对高脂源性骨髓间充质干细胞内皮分化的影响及其潜在机制

Ox-LDL inhibits endothelial differentiation of bone marrow mesenchymal stem cells in rats

  • Received:2013-11-18 Revised:2014-04-18 Online:2015-01-05 Published:2014-12-30
  • Contact: Tiewei Lv E-mail:ltw200145@163.com

摘要: 目的 观察氧化低密度脂蛋白(ox-LDL)对大鼠骨髓间充质干细胞(MSCs)内皮分化的影响及其机制。 方法 将可内皮分化的MSCs分为对照组(不加任何干预因素),Ox-LDL组(加入5μg/mL的ox-LDL),ox-LDL+乙酰半胱氨酸组(n-acetylcycteine,NAC)预处理后再加等浓度ox-LDL), nLD对照组(加入5μg/mL的天然低密度脂蛋白 (native LDL,nLDL) 。光镜下观察细胞形态,免疫组化、Western blot及RT-PCR技术检测特异性内皮表面标记,流式细胞技术检测分化效率,电子顺磁震荡技术检测氧化活性产物(reactive oxygen species,ROS),Western blot技术测定细胞信号通道蛋白Akt。结果 1) MSCs能够分化为内皮细胞,表现为内皮表面标志vWF、Flk-1和CD31的出现和血管样结构的形成; 2) Ox-LD明显减弱MSCs向内皮分化(P<0.05),而NAC预处理后MSCs内皮分化能力恢复;3) ox-LDL干预后ROS明显升高(P<0.05),而NAC预处理后ROS产量明显降低(P<0.05);4. ox-LDL干预后磷酸化Akt表达明显降低(P<0.01),而NAC预处理后有所恢复,但仍低于正常培养组。结论 ox-LDL抑制大鼠MSCs向内皮分化, NAC可部分或完全反转ox-LDL的抑制效应,Akt发挥一定作用。

关键词: 氧化低密度脂蛋白, 骨髓间充质干细胞, 内皮细胞, 分化

Abstract: Objective: To investigate that Ox-LDL inhibits endothelial differentiation of bone marrow mesenchymal stem cells (MSCs) in rats and its underlying mechanism. Methods: Cultured MSCs were divided into four groups: blank groups, Ox-LDL groups (5μg/ml Ox-LDL), Ox-LDL+NAC groups (5μg/ml Ox-LDL and pre-treated NAC), and negative groups (5μg/ml nLDL). Cellular morphology, endothelial Marker and differentiation efficiency as well as signal of oxidative and pathway protein were detected after induction by Western blot, real–time PCR. Results: MSCs can differentiate into endothelial cells with expression of endothelial marker vWF,Flk-1 and CD31 at the mRNA and protein level and appearance of vascular morphology, ox-LDL obvious weaken endothelial differentiation of MSCs (P<0.05), but NAC can reverse the inhibited effect of ox-LDL, the amount of ROS in Ox-LDL groups was more higher than that in Ox-LDL+NAC groups(P<0.05), The expression of phosphorylated Akt became decreased distinctly after treatment with ox-LDL(P<0.05), NAC can restore its expression close to normal. Conclusion:ox-LDL can inhibit endothelial differentiation of MSCs via ROS, NAC restore partly or completely inhibited effect of ox-LDL,Akt signaling also played a critical role.

Key words: oxidized low density lipoprotein, Rat bone marrow mesenchymal stem cells, endothlial cells, differentiation.

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