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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。
AIM: To investigate the effect of murine double minute 2 (MDM2) antisense oligonucleotide (ASON) on the expression of MDM2 and p53 in vascular smooth muscle cells (VSMCs), and to explore the role of MDM2 antisense oligonucleotide The feasibility of embedding scaffold in the prevention and treatment of stent restenosis. Methods: Antisense oligodeoxynucleotides targeting MDM2 mRNA were synthesized and transfected into rabbit vascular smooth muscle cells with different concentration of ASON. RT-PCR and Western-blotting were used to detect the effect of MDM2 antisense oligonucleotide on the proliferation of vascular smooth muscle cells MDM2 and p53 expression. Results: After MDM2 antisense oligonucleotides were treated with different concentrations of vascular smooth muscle cells, the expressions of MDM2 and p53 mRNA in each group were significantly different (P <0.01). The expression of MDM2 and p53 protein in each concentration group There was a significant difference (P <0.01). CONCLUSION: MDM2 antisense oligonucleotide can specifically inhibit the MDM2 expression in rabbit vascular smooth muscle cells and increase the expression of p53 gene in vitro. MDM2 antisense oligonucleotide is expected to be further used in drug-eluting stent studies.