三氧化二砷对大鼠血管平滑肌细胞增殖与凋亡的影响

The effects of arsenic trioxide on the proliferation and apoptosis of vascular smooth muscle cells of rats

目的观察三氧化二砷(As2O3)对大鼠血管平滑肌细胞(VSMC)增殖及凋亡的影响,以提供含As2O3缓释血管内支架植入手术后防止再狭窄的实验依据。方法分离培养大鼠VSMC并以As2O3处理;用四甲基偶氮唑蓝还原反应(MTT法)检测细胞增殖情况,RT-PCR检测增殖细胞核抗原(PCNA)mRNA;以AnnexinV+PI双染色流式细胞术检测细胞凋亡;比色法检测半胱氨酸天门冬氨酸蛋白酶-3(Caspase-3)的活性。结果 2μmol/L的As2O3对VSMC的生长有明显的抑制作用,且此作用呈时间依赖性(P<0.01)。以2μmol/L的As2O3作用72h后,AS2O3对PCNAmRNA表达有显著的抑制作用(P=0.009),对VSMC凋亡的发生(P=0.0001)及对Caspase-3的激活(P=0.005)均有显著促进作用。结论 As2O3可有效抑制VSMC增殖,诱导VSMC凋亡。As2O3的应用有可能为防治血管内支架植入术后再狭窄的药物研究提供一个新思路。

Objective To observe the effects of arsenic trioxide(As2O3)on the proliferation and apoptosis of vascular smooth muscle cells(VSMC)of rats in order to investigate the possibility for its application in slow-release drug delivery endovascular stent.Methods VSMCs of rats were isolated,cultured and treated with As2O3;methyl thiazolyl tetrazolium reduction reaction(MTT method)was used to measure the proliferation of cell,and the mRNA of proliferating cell nuclear antigen(PCNA)was measured by RT-PCR;the apoptosis was measured using Annexin V and PI double staining by flow cytometry;colorimetric method was employed to measure the activities of cysteine aspartic acid protease-3(Caspase-3).Results 2 μmol/L As2O3 showed significant inhibitory effects on the growth of VSMC(P<0.01 in comparison with the control group),and this effects were time-dependent.As2O3 showed significant inhibitory effects on the expression of PCNA mRNA after the cells were treated with 2 μmol/L As2O3 for 72 h(P=0.009),and it did significantly promote the apoptosis of VSMC(P=0.0001)and the activation of Caspase-3(P=0.005).Conclusion As2O3 can effectively inhibit the proliferation of VSMC and induce the apoptosis of VSMC.The application of As2O3 may provide a new idea for the investigations on drugs for the prevention and treatment on restenosis after endovascular stent implantation.