GRP78在apoE基因缺失小鼠脑、肝脏及主动脉中的基因表达分析

【作者】 任莉萍；

【导师】 潘杰；

【作者基本信息】 山东师范大学 ， 细胞生物学， 2007， 硕士

【摘要】 动脉粥样硬化(atherosclerosis,AS)是心脑血管系统中最常见的疾病,是冠心病、心肌梗塞等心脑血管病的物质基础,严重危害着人类健康,在西方发达国家一直是主要的死亡原因之一。近年来,本病在我国亦有明显增加,其发病率和死亡率逐年上升,并呈年轻化趋势。因此,对AS的研究日益重要。载脂蛋白E基因缺失(apoE-/-)小鼠的AS形成呈自发性、进展性、连续性、广泛性,其病变程度与人AS形成相似,具有代表性,是探讨AS发生发展的优良模型。葡萄糖调节蛋白(GRP)78是在应激状态下细胞内质网明显高表达的一类应激蛋白,是内质网应激的标志分子,与HSP70有很高的同源性,是HSP70家族的重要组成部分。内质网应激是细胞受到刺激时的一种重要的防御机制,可以提高细胞的应激耐受力,延长细胞在各种有害刺激下的存活期。本文以apoE-/-小鼠为实验动物模型,首先研究了apoE-/-小鼠早期AS病变特征,进而探讨了GRP78在apoE-/-小鼠中的基因表达情况。结果发现:1. apoE-/-小鼠血浆TC、TG、LDL-C水平均高于同龄野生型小鼠,并随年龄的增长而升高。apoE-/-小鼠血浆TC和LDL-C均极显著高于野生型小鼠。2.光学显微镜观察显示:1月龄apoE-/-小鼠主动脉根部瓣膜处未见苏丹Ⅳ阳性染色区,2月龄时主动脉根部内膜出现轻微的苏丹Ⅳ阳性染色区,3月龄apoE-/-小鼠主动脉根部内膜出现明显的苏丹Ⅳ阳性染色区。野生型小鼠从1月龄至3月龄主动脉根部瓣膜处未发现明显苏丹Ⅳ阳性染色区。3.根据RT-PCR结果显示,GRP78在apoE-/-小鼠脑和动脉中从出生后14天至3月龄内表达有明显变化;但在肝脏中的表达没有明显变化。而GRP78在野生型小鼠的肝脏、脑和动脉中的表达则没有明显变化。更多还原

【Abstract】 Atherosclerosis (AS) is the most common cardiovascular system diseases. It is the materious foundation of the Coronary heart, myocardial infarction and other cardiovascular diseases, it seriously endanger human’s health. In Western countries, it has been one of the main causes of death. In recent years, there was significant increase in China. Its incidence and mortality rates rise year by year, younger by younger. Therefore, the reaserch of atherosclerosis has become more and more important.The AS formation of apolipoprotein E gene deleted mice (apoE-/-) was spontaneous, progressive, continous and extensive. To a large extent,it is similar to the AS formation of human, it is a reliable and representative model for the study of development of AS.Glucose-regulated protein (GRP) 78, a kind of stress proteins, will significantly high express under the stress in the endoplasmic reticulum. It is the marker of the endoplasmic reticulum stress with a high homology to HSP70. It was considered as an important component of HSP70 families. Endoplasmic reticulum stress is an important self-defense mechanism when the cells were stimulated. It can increase the stress tolerance of cell, extend its survival period when it was under the harmful stimulation.This paper is based on apoE-/- mice AS model. First, there is a study of early atherosclerotic lesions feature of apoE-/- mice. Furthermore, the expression of the gene GRP78 in apoE-/- mice is also explored.The results showed that the plasma TC, TG and LDL-C of apoE-/- mice were higher than the same age of wild-type mice, and increased with age. Plasma TC and LDL-C of the apoE-/- mice were significantly higher than those of wild-type mice.Optical microscopy results showed that there was no obvious regions stained with Sudan IV in the valvular of the aortic root when apoE-/- mice were 1 month old until it grew to 3 months, there were significant regions stainied with Sudan IV in the valvular of the aortic root. The wild-type mice from one month to three months old have not been found positive stained regions of Sudan IV in the valvular of the aortic root.The RT-PCR results show that the expression of the gene GRP78 in apoE-/- mice’s brain and artery was distinctive during 14 days and 3 month after birth.while it was not been found in apoE-/- mice’s liver tissue. Little obvious distinction was also found in wild-type mice from 14 days to3 month old.更多还原