摘要
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 RNA interference for A20 gene on the phenotype of Human Umbilical Artery Smooth Muscle Cells （HUASMC） induced by H2O2 and its possible molecular biology mechanism. Methods： HUASMC was randomly divided into six groups： （1） blank control group, （2） A20 siRNA group, （3） scramble siRNA group, （4） H2O2 group, （5） H2O2＋A20siRNA group, （6） H2O2 ＋ scramble siRNA group. RT-PCR and western-blotting were used to detect the change of expression of A20 gene. Transmission electron microscope was used to determine the change of ultra-structure of the cells. Results： The faint level expression of A20mRNA and protein was detected in control group, and depressed after siRNA transfection. The expression level of A20mRNA and protein increased significantly in H2O2 group. The expression level was lower in the control group than in the H2O2＋A20siRNA group （P〈0.05）and the interference efficiency was about 55%. Representative contractile phenotype of VSMC was shown in the control group and synthetic phenotype was shown in H2O2 group. The result in the H2O2＋A20siRNA group shows representative synthetic phenotype with less myofilament and more organelles and secretions. Conclusions： A20 gene inhibits HUASMC phenotype transformation that induced by H2O2.
出处
《现代生物医学进展》
CAS
2009年第11期2031-2034,共4页
Progress in Modern Biomedicine
基金
国家自然科学基金资助项目(30670836)
