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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(10ng/mL)、二甲双胍组(2mmol/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-9mRNA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。
Objective To investigate the effect of metformin on the migration of vascular smooth muscle cells (VSMCs) induced by TNF-α and explore its possible molecular mechanism. Methods The control group, TNF-α group (10ng / mL), metformin group (2mmol / L), metformin + TNF-α group (with the same concentration). The migration ability of cells was detected by scratch assay and transwell assay respectively. The expression of matrix metalloproteinase (MMP) -2 and MMP-9 mRNA was detected by RT-PCR and the protein expression of MMP-2 and MMP-9 As well as the expression of NF-κB and cytoplasmic IκB in each group. Results Scratch assay and transwell assay showed that the migration ability of TNF-α group was higher than that of the control group (P <0.01), and the migration ability of metformin group was lower than that of the control group (P <0.05). The migration ability of metformin + TNF- (P <0.01). The expression of MMP-2, MMP-9 and NF-κB in TNF-α group was higher than that in control group, while the expression of IκB was decreased (P <0.01). The expression of MMP-2, MMP-9 and NF-κB in metformin + TNF-α group was significantly lower than that in TNF-α group (P <0.01). The expression of MMP-2 and MMP-9 in metformin group was significantly lower than that in control group (P <0.05 or P <0.01), but there was no significant difference in the expression of NF-κB and IκB between control group and metformin group Statistical significance. Conclusion Metformin can inhibit the migration of VSMCs induced by TNF-α, and its inhibitory effect may be related to the down-regulation of the expression of matrix metalloproteinase and the block of NF-κB signaling pathway.