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目的观察吡格列酮(Pio)对胰岛素(Ins)刺激的血管平滑肌细胞(VSMC)增殖的干预作用,及对增殖核抗原(PCNA)基因和蛋白表达水平的影响。方法 (1)以四甲基偶氮唑盐比色法观察不同浓度、不同时间的Ins对VSMC增殖的影响,以及Pio和Ins共孵育对VSMC增殖的作用。(2)用半定量逆转录聚合酶链反应测定PCNA mRNA的表达。(3)用Western blot法检测PCNA蛋白的表达。结果Ins促进VSMC增殖,促增殖效应在72h末达峰值,差异有统计学意义(P<0.01);而Pio能抑制Ins诱导的VSMC增殖及PCNA mRNA和蛋白表达水平,最佳浓度100μmol/L(P<0.01)。结论吡格列酮抑制胰岛素的促VSMC增殖作用,通过降低PCNA的表达影响VSMC表型转换,有望用于治疗动脉粥样硬化。
Objective To investigate the effects of pioglitazone on the proliferation of vascular smooth muscle cells (VSMCs) stimulated by insulin and the effect of Pio on the expression of PCNA gene and protein. Methods (1) Tetramethylzirconate colorimetry was used to observe the effects of Ins at different concentrations and different times on the proliferation of VSMCs, and the effect of co-incubation of Pio and Ins on the proliferation of VSMCs. (2) The expression of PCNA mRNA was determined by semi-quantitative reverse transcription polymerase chain reaction. (3) The expression of PCNA protein was detected by Western blot. Results Ins increased the proliferation of VSMC and reached its peak value at the end of 72 h (P <0.01). However, Pio inhibited the proliferation of VSMC induced by Ins and the expression of PCNA mRNA and protein with the optimal concentration of 100 μmol / L P <0.01). Conclusion Pioglitazone inhibits the proliferation of VSMC induced by insulin, which may be used to treat atherosclerosis by reducing the expression of PCNA and affecting the phenotype of VSMC.