论文部分内容阅读
目的:通过用高糖对培养的大鼠主动脉平滑肌细胞进行干预,观察其对基质金属蛋白酶-2(MMP-2)表达的影响。方法:用组织贴块法体外培养大鼠主动脉平滑肌细胞,分组后分别予以葡萄糖0,1,5g/L进行干预。分别提取细胞总RNA和总蛋白分别进行逆转录PCR(RT-PCR),Westernblotting检测,同时取出条件培养基经酶谱法(zymography)检测分泌于培养基中的MMP-2活性。结果:经mRNA,蛋白和酶活性检测,均发现在葡萄糖浓度为5g/L时,MMP-2的表达水平较对照和1g/L时明显增加。结论:高糖促进血管平滑肌细胞和内皮细胞内MMP-2表达,提示血管细胞外基质的降解在糖尿病患者动脉粥样硬化发生机制中起着一定作用。
OBJECTIVE: To investigate the effects of high glucose on the expression of matrix metalloproteinase-2 (MMP-2) in rat aortic smooth muscle cells. Methods: Rat aorta smooth muscle cells were cultured in vitro by tissue patch method. The cells were treated with 0, 1 and 5 g / L glucose respectively. The total RNA and total protein of the cells were respectively extracted for reverse transcription PCR (RT-PCR) and Western blotting. Meanwhile, the conditioned medium was removed and the secretion of MMP-2 in the medium was detected by zymography. Results: The results of mRNA, protein and enzyme activity showed that the expression of MMP-2 was significantly increased at glucose concentration of 5 g / L compared with the control and 1 g / L. Conclusion: High glucose can promote the expression of MMP-2 in vascular smooth muscle cells and endothelial cells, suggesting that degradation of vascular extracellular matrix plays a role in the pathogenesis of atherosclerosis in diabetic patients.