P53基因半剂量缺失对活性维生素D缺乏小鼠肾脏的影响

发布时间：2018-03-21 13:26

本文选题：维生素D　切入点：p53　出处：《中南大学》2014年硕士论文　论文类型：学位论文

【摘要】：研究表明：维生素D缺乏／不足是慢性肾脏病的共同特征,同时,维生素D缺乏也加剧了慢性肾脏疾病的进展,并作为独立的危险因素直接影响着正常人群及慢性肾脏病患者的死亡率。我们在敲除1α羟化酶基因造成内源性1,25(OH)2D3缺乏的小鼠模型(1α(OH)ase-/-)中观察到小鼠的多器官的早衰模型(可见小鼠生长发育滞后、皮肤变薄、皮下脂肪减少、骨质疏松、肾功能减退、肾间质纤维增多等)及多种慢性疾病的发生(包括慢性肾脏病)。而有“基因组卫士”之称的p53基因对活性维生素D缺乏的肾脏有何种影响及其影响机制尚不清楚。为了解答上述疑问,我们建立了p53基因半剂量缺失小鼠模型p53+/-,并将其和1α(OH)ase+/-小鼠雌雄交配获得的p53+/-1α (OH)ase+/-双基因小鼠,由p53+/-1α(OH)ase+/-基因小鼠雌雄交配获得同窝的1α(OH)ase-/-、WT(野生型)、p53+/-及p53+/-1α (OH)ase-/-四种基因型的雄性小鼠,同时辅以高钙、磷饲料至10周龄,排除血清钙、磷高低的不同的影响。同时我们分别运用形态学、组织化学、免疫组织化学、Western Blot、Masson染色、流式细胞术等方法,比较分析四种基因型小鼠肾脏差异。通过应用以上方法的比较分析,我们发现p53基因能在不同程度上改善因活性维生素D缺乏引起的小鼠体型、体重及肾脏大小及肾脏重量与体重的比值的下降、血清肌酐、尿素氮等肾功能指标的上升、ROS水平的升高、肾脏内细胞增殖指标(如PCNA、Ki67等)表达水平的下降、肾脏的间质纤维的增加、细胞DNA损伤增加等。以上的结果证实,p53基因对活性维生素D缺乏的小鼠肾脏具有保护性的作用,同时也为p53基因应用于防治肾脏疾病提供了实验和理论依据。
[Abstract]:Studies have shown that vitamin D deficiency / deficiency is a common feature of chronic kidney disease and that vitamin D deficiency exacerbates the progression of chronic kidney disease. As an independent risk factor, the mortality of normal population and patients with chronic kidney disease was directly affected. We observed the preterm senescence of multiple organs in mice model of 1 伪 -hydroxylase gene knockout resulting in endogenous 1 1 OHH 2D3 deficiency. The model (which shows that the growth and development of mice is lagging), Skin thinning, subcutaneous fat loss, osteoporosis, renal dysfunction, The pathogenesis of various chronic diseases (including chronic kidney disease) and p53 gene, known as "genome defender", has not been clear on the kidney with active vitamin D deficiency and its mechanism. To answer the above questions, We established a half dose deletion mouse model of p53 gene p53 / -, and then we mated it with 1 伪 -OHHase / -mouse male and female mice to get p53 / 1 伪 -OHHase / a-double gene mice. Male mice with four genotypes of p53- and p53 / -1 伪 -OHase / -gene mice were obtained by mating with male and female mice with the same litter of 1 伪 -OHH- 伪 -p5- and p53 / -1 伪, supplemented with high calcium and phosphorus forage to the age of 10 weeks, to exclude serum calcium, the male mice with four genotypes of p53- and p53- 1 伪 -OHase-r-1 were obtained by mating with the female and male mice with the gene p53 / 1 伪. At the same time, we compared and analyzed the renal differences among the four genotypes by means of morphological, histochemical, immunohistochemical Western BlotMasson staining and flow cytometry. By comparing and analyzing the above methods, we found that p53 gene could improve the body size, kidney size and the ratio of kidney weight to body weight, serum creatinine in mice caused by active vitamin D deficiency in varying degrees. The increase of renal function, such as urea nitrogen, the increase of Ros level, the decrease of cell proliferation index (such as PCNAnKi67, etc.) and the increase of interstitial fiber in kidney. The above results confirmed that p53 gene had protective effect on kidney of mice with active vitamin D deficiency and provided experimental and theoretical basis for the application of p53 gene in the prevention and treatment of renal diseases.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R692

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