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目的通过定量分析雌激素受体在动脉粥样硬化兔心脏和肝脏内的相对基因表达,研究雌激素受体基因mRNA的表达水平与动脉粥样硬化(AS)发生的关系。方法手术法去势新西兰雌兔,高胆固醇饮食法复制AS模型,以管家基因-βactin作为内源性校准基因,用SYBR Green I作为荧光染料,采用实时荧光定量逆转录聚合酶链式反应(real-time RT-PCR)对ERαmRNA的表达进行了分析。结果对照组(N)、模型组(C)和假手术组(SC)主动脉硬化斑块面积分别为0、(0.75±0.24)和(0.56±0.27),饮食法复制AS模型成功;ER在对照组兔心脏和肝脏内的相对基因拷贝数分别为0.46、0.49,在模型组的去势雌兔心脏和肝脏内的相对基因拷贝数分别为0.29、0.32,在假手术组兔心脏和肝脏内的相对表达量分别为0.53、0.51。结论兔心脏、肝脏内都有雌激素受体α分布;去势AS雌兔组织内雌激素受体表达量的减少可能与动脉硬化的形成有关。
Objective To study the relationship between the expression of estrogen receptor mRNA and the development of atherosclerosis (AS) by quantitative analysis of the relative gene expression of estrogen receptor in the heart and liver of atherosclerotic rabbits. Methods The ovariectomized New Zealand female rabbits were enrolled in this study. High cholesterol diet was used to replicate the AS model. The housekeeping gene-βactin was used as the endogenous calibration gene. SYBR Green I was used as the fluorescent dye. Real-time fluorescence quantitative reverse transcription polymerase chain reaction -time RT-PCR) was used to analyze the expression of ERa mRNA. Results The area of aortic plaque in control group (N), model group (C) and sham operation group (SC) were 0, 0.75 ± 0.24 and 0.56 ± 0.27, respectively. In the control group, the relative gene copy numbers in the heart and liver were 0.46 and 0.49, respectively. The relative gene copy number in the heart and liver of the model group was 0.29 and 0.32, respectively. The relative expression levels were 0.53,0.51. Conclusion The distribution of estrogen receptor α in the heart and liver of rabbits may be related to the decrease of estrogen receptor expression in the ovariectomized female rabbits, which may be related to the formation of arteriosclerosis.