| 彭炬 熊丹 马晓东 兰时乐 卢向阳.miR-375 在AGEs介导糖尿病血管损伤细胞中的生物学功能[J].,2014,14(13):2441-2445 |
| miR-375 在AGEs介导糖尿病血管损伤细胞中的生物学功能 |
| The Biological Function of miR-375 in Vascular Cells Injury Induced byAGEs-mediated Diabetes |
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| DOI： |
| 中文关键词: miR-375 AGEs 转染 血管损伤细胞 |
| 英文关键词: miR-375 AGEs Transfection Injured vascular cells |
| 基金项目: |
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| 中文摘要: |
| 目的：探讨miR-375 在血管损伤细胞中的表达及生物学功能。方法：利用基因克隆技术构建miR-375表达载体；然后将
miR-375 表达质粒转染至血管损伤细胞中，同时分别设立Huvec12对照组，血管损伤细胞组，血管损伤抑制组，Huvec12 转染
miR-375 组。24h 后收集细胞，在mRNA 和蛋白水平检测Mtpn、NFγB、profilin1、sICAM1 的表达，经荧光染色观察细胞F-actin 的
变化，再用流式细胞仪检测细胞凋亡。结果：血管损伤细胞中过表达miR-375后，在mRNA和蛋白水平靶基因Mtpn 下降，NFγB
的表达活性下降，使糖尿病血管病变的标志profilin1 下调；F-actin 表达恢复；细胞粘附因子（sICAM1）表达下降，细胞凋亡减少。
结论：初步证明miR-375 可以抑制AGEs 介导的糖尿病血管细胞损伤的发生，可能成为糖尿病血管损伤并发症基因治疗的靶点。 |
| 英文摘要: |
| Objective:To investigate the expression and biological function of miR-375 in injured vascular cells.Methods:Cloning technology was used to construct miR-375 expression vector and subsequently mir-375 expression plasmid was transfected into
injured vascular cells. Meanwhile, we set for further study: Huvec12 as control, injured vascular cells, vascular cells after injury inhibition
and cells with miR-375 overexpression. Cells were harvested 24 hours after treatments. The expression of Mtpn, NFγB, profilin1 and
sICAM1 were measured in both mRNA and protein level. F-actin was examined via fluorescence staining. Cell apoptosis was assessed by
flow cytometry.Results:In injured vascular cells with miR-375 overexpression, the mRNA and protein expression of targeted gene Mtpn
declined. It was also observed the reduction of NFγB activity, downregulation of the diabetic vascular injury marker profilin1, restoration
of F-actin expression, a decrease of sICAM1 expression and reduced cell apoptosis.Conclusion:miR-375 may inhibit the injuries of
vascular cells caused by AGEs-mediated diabetes and may become a new target for gene therapy to treat vascular complications caused
by diabetes. |
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