| 摘要: |
| 目的 探讨二甲双胍对TNF-α诱导的血管平滑肌细胞（vascular smooth muscle cells，VSMCs）迁移的影响并探讨其可能的分子机制。方法 实验设对照组、TNF-α组（10 ng/mL）、二甲双胍组（2 mmol/L）、二甲双胍+TNF-α组（浓度同前）。分别用划痕实验和侵袭小室（transwell）实验检测细胞的迁移能力；RT-PCR法检测基质金属蛋白酶（MMP）-2、MMP-9 mRNA的表达；蛋白免疫印迹法检测MMP-2、MMP-9以及各组细胞核内NF-κB、胞质IκB的表达。结果 划痕实验和transwell实验均显示TNF-α组细胞迁移能力高于对照组（P<0.01），二甲双胍组细胞迁移能力低于对照组（P<0.05），二甲双胍 TNF-α组细胞迁移能力低于TNF-α组（P<0.01）；TNF-α组MMP-2、MMP-9、NF-κB的表达高于对照组，IκB的表达则降低（P<0.01）。二甲双胍 TNF-α组较TNF-α组MMP-2、MMP-9、NF-κB的表达则明显下降，IκB的表达则升高（P<0.01）。二甲双胍组细胞内MMP-2、MMP-9的表达与对照组比较差异有统计学意义（P<0.05或P<0.01），但细胞内NF-κB、IκB蛋白的表达与对照组相比差异无统计学意义。结论 二甲双胍能抑制TNF-α诱导的VSMCs迁移，其抑制作用可能与基质金属蛋白酶表达量的下调及NF-κB信号通路的阻断有关。 |
| 关键词: 二甲双胍 肿瘤坏死因子α 血管平滑肌细胞 细胞迁移 |
| DOI：10.3724/SP.J.1008.2014.01029 |
| 投稿时间：2014-03-06修订日期：2014-05-22 |
| 基金项目: |
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| Effect of metformin on TNF-α-induced migration of vascular smooth muscle cells and the underlying mechanism |
| ZHANG Duo-duo,LI Pai,ZHANG Wei,KONG Jing-jing,QIN Yu,WEI Chun-yang* |
(Department of Special Medicine, The First Affiliated Hospital of Dalian Medical University, Dalian 116011, Liaoning, China
* Corresponding author.) |
| Abstract: |
| Objective To explore the effect of metformin on TNF-α-induced migration of vascular smooth muscle cells (VSMCs) and the possible mechanism. Methods VSMCs were divided into four groups: control group, TNF-α group(10 ng/mL), metformin group(2 mmol/L) and metformin+TNF-α group(2 mmol/L 10 mg/mL). The migration of VSMCs was detected by wound healing assay and transwell assay. The expressions of matrix metalloproteinase (MMP)-2 and MMP-9 were detected by RT-PCR and Western blotting analysis; the protein expression of nuclear NF-κB and cytoplasmic IκB were also determined by Western blotting analysis. Results Wound healing assay and transwell assay showed that the migration ability of VSMCs in TNF-α group was significantly increased compared with the control group (P<0.01), while that of metformin TNF-αgroup was significantly decreased (P<0.05). Compared with TNF-α group, the migration ability of metformin TNF-α group was significantly decreased (P<0.01). MMP-2, MMP-9 and NF-κB expression levels were significantly increased and the IκB expression was significantly decreased in TNF-α group compared with those in the control group (P<0.01). Compared with TNF-α group, MMP-2, MMP-9 and NF-κB expression levels were significantly decreased and IκB expression was significantly increased in metformin TNF-α group (P<0.01). The intracellular expression levels of MMP-2 and MMP-9 were significantly different between metformin group and control group (P<0.05 or P<0.01), while the intracellular NF-κB, IκB proteins were not significantly different. Conclusion It suggests that metformin can effectively inhibit the TNF-α-induced migration of VSMCs, which might be related to the down-regulation of matrix metalloproteinases and the blockage of the NF-κB signal pathway. |
| Key words: metformin tumor necrosis factor-α vascular smooth muscle cells cell migration |
