| 摘要: |
| 目的 探索3-甲基腺嘌呤(3-MA)对大鼠骨髓源性内皮祖细胞(EPCs)超微结构和自噬体膜型(LC3-II)蛋白表达的 影响。方法 采用密度梯度法取大鼠骨髓EPCs,体外诱导分化并鉴定;分成对照组和4个3-MA浓度组(分别加入1.25、2.5、5、 10mmol/L 3-MA) 。采用单丹(磺)酰戊二胺(MDC)荧光染色和透射电镜观察 3-MA 给药后 EPCs 自噬的发生。Western blot 检测 3-MA给药后EPCs内LC3-II蛋白表达的变化。结果 MDC荧光染色和透射电镜均观察到5、10mmol/L组EPCs超微结构明显 区别于正常及死亡细胞,是典型的凋亡形态图。Western blot 检测 3-MA 给药 24h 后 EPCs LC3-II 蛋白表达降低;与对照组比较,5、 10mmol/L组LC3-II蛋白表达明显降低(P<0.05)。结论 5、10mmol/L的3-MA对EPCs超微结构和LC3-II蛋白表达均有影响, 推测浓度≥5mmol/L 的 3-MA 能抑制 EPCs 的自噬。 |
| 关键词: 内皮祖细胞 单丹(磺)酰戊二胺 3-甲基腺嘌呤 |
| DOI： |
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| 基金项目:国家自然科学基金资助项目 |
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| Effect of 3-methlyadenineon ultrastructure and LC3-II expression of rat endothelial progenitor cells |
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ZHU Renda, LI Xiaoqiang
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the Second Affliated Hospital of Soochow University，Suzhou
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| Abstract: |
| Objective To investigate the effect of 3-methlyadenine(3-MA ) on ultrastructure and LC3-II protein expression in rat bone marrow-derived endothelial progenitor cells (EPCs). Methods Mononuclear cells (MNCs) were isolated from rat bone marrow; the EPCs were induced from cultured MNCs and identified.The induced rat EPCs were treated with different concentrations of 3-MA (0, 1.25, 2.5, 5, 10mmol/L). Monodansylcadaverine staining and transmission electron microscopy (TEM) were used to examine cell autophagy induced by 3-MA in rat EPCs.Western blot analysis was used to detect the LC3-II protein expression. Results Autophagy inhibition was observed in EPCs by 3-MA at concentration of 5mmol/L and 10mmol/L by both MDC staining and TEM. LC3-II protein expression in EPCs was decreased aftertreatment of 5mmol/L and 10mmol/L 3-MA as detected by western blot (P＜0.05). Conclusion High dose of 3-MA caninduce the ultrastructural changes and reduce LC3-II expression in rat bone marrow-derived epithelial progenitor cells. |
| Key words: Endothelial progenitor cell Monodansylcadaverine 3-methlyadenine |
