摘要
目的以动脉粥样硬化(AS)的独立危险因子同型半胱氨酸(Hcy)处理人脐静脉血管平滑肌细胞(VSMCs),观察Hcy对VSMCs多价核因子CTCF表达的影响,探讨Hcy致AS发病的可能机制。方法 将培养的VSMCs分为0、50、100、200、500及1000 mol/L Hcy处理组,以0 mol/L Hcy组为空白对照。在VSMCs覆盖瓶底70%时,将不同浓度的Hcy加入培养液,继续培养VSMCs 48 h,采用半定量逆转录聚合酶链反应(RT-PCR)法和免疫组织化学方法分别检测VSMCsCTCF的mRNA和蛋白表达水平。结果与对照组比较,Hcy刺激48h后,VSMCs CTCF的mRNA和蛋白表达皆明显增加,以高浓度组为著(500、1000 mol/LHcy,<0.05)。CTCF免疫组织化学阳性染色以细胞核为主。结论 Hcy升高可诱导VSMCs CTCF的表达上调。
Objective To investigate the potential impact of homcysteine(hcy) on the expression of CCCTC-binding factor(CTCF) in VSMCs and its possible role in the pathogenesis of atherosclerosis(AS).Methods Cultured human umbilical VSMCs were divided into 0,50,100,200,500,1000 %emol/L Hcy groups,the 0 %emol/L Hcy group was the control.After the VSMCs were treated with different concentrations of Hcy for 48h,the mRNA level of CTCF was assayed by reverse transcription polymerase chain reaction(RT-PCR),and its protein expression were detected by immunohistochemistry.Results After Hcy treatment,the level of CTCF mRNA in VSMCs was increased,especially in 500,1000 %emol/L Hcy groups(P <0.05).Immunohistochemistry showed that the CTCF positive staining of VSMCs in control group was very weak,but in 500,1000 %emol/L Hcy groups,the CTCF staining was significantly increased,the positive staining was mainly concentrated in the area of nucleus.Conclusions Hcy can induced an up-expression of CTCF in VSMCs.
出处
《遵义医学院学报》
2010年第6期528-534,534,共4页
Journal of Zunyi Medical University
基金
贵州省科技厅(No.黔科合J字[2010]2277)
遵义医学院博士启动基金(F-460)
