目的:研究小鼠双微体扩增基因(mouse double minute 2；MDM2)反义寡核苷酸(antisense oligonucleotide; ASON)对血管平滑肌细胞MDM2和p53表达的影响,探讨MDM2反义寡核苷酸包埋支架防治支架内再狭窄的可行性.方法:人工合成一段针对MDM2 mRNA的反义寡核苷酸,脂质体包裹不同浓度ASON转染兔血管平滑肌细胞,RT-PCR和Western-blotting检测MDM2反义寡核苷酸对兔血管平滑肌细胞MDM2和p53表达的影响.结果:不同浓度MDM2反义寡核苷酸作用于兔血管平滑肌细胞后,MDM2和p53 mRNA表达量各浓度组之间有显著性差异(P＜0.01),MDM2和p53蛋白表达量各浓度组之间有显著性差异(P＜0.01).结论:MDM2反义寡核苷酸体外能够特异性抑制兔血管平滑肌细胞MDM2表达,提高细胞内p53基因表达量,MDM2反义寡核苷酸有望被进一步应用于药洗脱支架研究.%Objective: To investigate the possibility of using MDM2 mRNA antisense oligonucleotide (ASON) on preventing restenosis following angioplasty, the effection of the MDM2 ASON on MDM2 and p53 was studied in vascular smooth muscle cells. Methods: An ASON targeted to MDM2 mRNA were synthesized. RT-PCR and Western-blotting were carried out to assay the MDM2 and p53 mRNA and protein level after vascular smooth muscle cells were transfected with liposome-coated ASON. Results: The mRNA level of MDM2 and p53 had significant different at different concertration of ASON (P<0.01). The protein level of MDM2 and p53 were significant different at different concertration of ASON (P<0.01). Conclusion: The MDM2 ASON can be specially hybridized to the MDM2 mRNA and inhibit MDM2 gene expression and increase p53 gene expression in vascular smooth muscle cell in vitro which provide a basis for it to be used in drug eluting stent study.
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