摘要
目的研究罗格列酮对高糖诱导的大鼠血管平滑肌细胞(VSMC)生长的影响,从而探讨其对糖尿病血管并发症的作用及机制。方法利用组织贴块法体外培养SD大鼠主动脉血管平滑肌细胞并分组给药,采用MTT比色法检测细胞生长活性;用W estern b lotting检测细胞增殖核抗原的表达;以流式细胞仪检测细胞周期的进程;RT-PCR检测MMP-2 mRNA的水平。结果罗格列酮呈浓度依赖性抑制高糖诱导的大鼠VSMC的增殖;降低PCNA的表达(P<0.05),明显降低细胞的S期数目百分比(P<0.01),增加G0/G1期数目百分比(P<0.01);明显降低MMP-2 mRNA(P<0.01)的表达。结论从细胞分子水平说明罗格列酮可能通过降低PCNA的表达,阻止细胞进入S期,减少有丝分裂来发挥对高糖诱导的VSMC增殖的抑制作用,MMP-2在罗格列酮改善血管重构的过程中可能发挥了重要的作用。
Objective: To investigate the effect of Rosiglitazone (RSG) on the proliferation of rat aortic vascular smooth muscle cells (VSMCs) induced by high glucose and the possible mechanism involved in the effect. Methods: The smooth muscle cells were cultured from the thoracic aorta of Sprague-Dawley (SD) rat. The viability of VSMCs was determined by3-(4, 5-dimethy 1-2-thiazoly) 2,5-diphenyl-2H-tetyazolium bromide (MTT) assay. The cells cycle was examined by flow cytometry. The protein expression of proliferating cell nuclear antigen (PCNA) in VSMCs was evaluated by Western blotting. The mRAN expression of Matrix metalloproteinases-2 (MMP-2) of VSMC was determined by RT-PCR. Results: Rosiglitazone at the concentration of 10μmol/l remarkedly inhibited the viability of VSMCs (P 〈 0.01 ), the expression of PCNA(P 〈 0.05)and the level of MMP-2 mRNA(P 〈 0.01 ) in VSMCs. Meanwhile, RSG decreased the proportion of VSMC "S" periods (P 〈 0.01 ) but increased the proportion of VSMCs G0/G1 periods ( P 〈 0.01 ). Conclusion : These findings suggest that RSG has its inhibiting effect on high glucose-induced VSMCs proliferation, at least partly by the way of halting the cell cycles and inhibiting PCNA expression and MMP-2 synthesis.
出处
《泰山医学院学报》
CAS
2008年第5期321-325,共5页
Journal of Taishan Medical College