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AGEs-RAGE通路阻断对血管平滑肌细胞迁移能力的影响

Effect of block of AGEs-RAGE pathway on the migration of VSMCs

  • 摘要: 研究AGEs-RAGE通路阻断对糖尿病大鼠主动脉血管平滑肌细胞(VSMCs)迁移能力的影响并探讨其可能机制。以晚期糖基化终产物受体(RAGE)抗体预孵VSMCs后,再用晚期糖基化终产物(AGEs)刺激VSMCs细胞,Transwell法检测VSMCs迁移能力变化,MTT法及ELISA分别检测VSMCs增殖及p27蛋白表达的变化,DCFH法测定VSMCs细胞内活性氧(ROS)水平,RT-PCR法测定NOX1 mRNA表达水平。结果显示:RAGE抗体可以明显抑制AGEs诱导的VSMCs迁移作用(P<;0.01),且抑制迁移作用较其抑制增殖作用强,细胞ROS水平降低(P<;0.01),NOX1 mRNA表达量下降但P27蛋白表达变化不大。实验结果表明,用RAGE抗体阻断AGEs-RAGE通路可以抑制VSMCs的迁移,其机制可能与其下调NOX1 mRNA表达从而降低细胞内氧化应激水平有关。

     

    Abstract: To investigate the effect of block of AGEs-RAGE pathway on the migration of aortic vascular smooth muscle in diabetic rats and its possible mechanisms, vascular smooth muscle cells(VSMCs)cells were pre-stimulated by antibody of RAGE, and then stimulated by AGEs. Transwell assay was adopted to assay migration of VSMCs. Proliferation of VSMCs and expression of p27 were analyzed by MTT and ELISA, respectively. The change of ROS level in VSMCs was defermined by DCFH assay, the expression of NOX1 mRNA was determined by RT-PCR assay. Results indicated that the AGEs induction for migration of VSMCs was significantly inhibited after treatment by RAGE antibody(P< 0. 01), which blocked the AGEs-RAGE pathway, and the inhibition of migration was stronger than that of proliferation. The ROS level was decreased(P< 0. 01), and the expression of NOX1 mRNA was decreased, yet the expression of P27 protein was not changed greatly. Block of AGEs-RAGE pathway by antibody of RAGE can inhibit the migration of VSMCs, and the mechanism may be related with the decrease of NOX1 mRNA and then down to the level of intracellular oxidative stress in VSMCs.

     

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