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目的:通过采用锌指蛋白A20基因干扰技术观察其对H2O2诱导的人脐动脉血管平滑肌细胞(HUASMC)超微结构影响,以初步研究锌指蛋白A20在氧化还原方面对细胞的保护作用。方法:将培养的人脐动脉血管平滑肌细胞随机分六组,空白组;A20siRNA组;scramble siRNA组;H2O2组;H2O2+A20siRNA组;H2O2+scramble siRNA,采用RT-PCR,Western-blotting方法观察A20基因表达变化,应用透射电镜观察细胞超微结构改变。结果:空白组A20mRNA与蛋白有微弱表达,A20siRNA组较空白组表达减少(P<0.05);H2O2组A20mRNA与蛋白表达明显高于空白组但显著低于H2O2+A20siRNA组(P<0.05)干扰效率大约55%。空白组平滑肌细胞(VSMC)电镜下呈典型的“收缩表型”;H2O2组VSMC表型发生明显改变,呈“合成表型”H2O2+A20siRNA组成典型的合成表型且肌丝明显减少,细胞器及分泌物增多。结论:A20基因可以抑制H2O2损伤诱导的血管平滑肌细胞表型转化。
OBJECTIVE: To observe the effect of zinc finger protein A20 gene interference on the ultrastructure of human umbilical artery smooth muscle cells (HUASMC) induced by H2O2 and to study the protective effect of zinc finger protein A20 on redox cells. Methods: The cultured human umbilical artery smooth muscle cells were randomly divided into six groups: blank group, A20 siRNA group, scramble siRNA group, H2O2 group, H2O2 + A20 siRNA group, H2O2 + scramble siRNA group and RT-PCR and Western- Changes in gene expression, transmission electron microscopy changes in the ultrastructure of cells. Results: A20 mRNA and protein were weakly expressed in A20 siRNA group and A20 siRNA group (P <0.05). A20 mRNA and protein expression in H2O2 group was significantly higher than that in blank group but significantly lower than that in H2O2 + A20 siRNA group (P <0.05) About 55%. VSMC showed a typical “contractile phenotype” under the electron microscope; the VSMC phenotype of H2O2 group changed obviously, which was a typical synthetic phenotype of H2O2 + A20siRNA with obvious myofilament Reduce, organelles and secretions increased. CONCLUSION: A20 gene can inhibit H2O2-induced vascular smooth muscle cell phenotype transformation.