胚胎时期肾脏对骨骼系统发育的相关性研究

发布时间：2018-09-09 08:56

【摘要】：目的:利用顺铂经肾脏排泄的特性,建立鸡胚胎肾脏损伤模型,将胚胎时期肾脏的发育与骨骼系统发育之间联系起来进行分析,以鸡胚为实验模型初步阐述胚胎时期肾脏与骨骼系统发育之间的关系。方法:将新鲜种鸡蛋48只,使用恒温培养箱孵化4d,在蛋壳上开窗,并随机分为4组进行空白对照和顺铂暴露:25μg/egg、50μlg/egg、100μg/egg组;继续孵化至12d时取胚胎进行评价;鸡胚发育到4.5d,分离胚胎肢芽组织,进行体外胚胎骨细胞培养;H.E及阿利新蓝-茜素红双染进行形态学观察;测定钙含量、25(OH)D及组织切片的碱性磷酸酶(ALP)和抗酒石酸酸性磷酸酶(TRAP)表达水平。结果:PBS和顺铂暴露随暴露剂量的增加,肾的发育在顺铂暴露组与PBS组之间存在显著性的差异,且呈现出剂量相关性(P0.01);Col 2al、ALP、Runx2、SOX9、MMP13、GADPH的RT-PCR结果显示,骨发生相关基因的表达降低(P0.05);25(OH)D及组织切片的ALP和TRAP随着顺铂暴露剂量的增加呈现相应的降低(P0.05)。结论:1)利用顺铂建立鸡胚胎肾损伤模型是可行的,肾损伤程度与剂量呈正相关关系;2)顺铂对鸡胚胎时期的骨骼发育没有直接影响;3)证实胚胎时期肾脏损伤影响骨骼发育,主要影响到成骨细胞和破骨细胞的功能;4)为部分先天性骨发育不良的发生提供一定的理论支持。
[Abstract]:Aim: to establish a chicken embryonic kidney injury model by using the characteristics of cisplatin excretion through the kidney, and to analyze the relationship between kidney development and skeletal system development in embryonic stage. Using chicken embryo as experimental model, the relationship between kidney and skeletal system development during embryonic period was discussed. Methods: 48 fresh chicken eggs were incubated in constant temperature incubator for 4 days. The eggs were randomly divided into 4 groups: blank control group and cisplatin exposure 25 渭 g / eggshell 50 渭 lg/egg,100 渭 g/egg group. The embryonic limb bud tissue was isolated from the chicken embryo at 4.5 days. The embryo bone cells were cultured in vitro with H. E and Alicin-alizarin red double staining for morphological observation. The expression levels of alkaline phosphatase (ALP) and tartrate-resistant acid phosphatase (TRAP) in tissue sections and calcium content of 25 (OH) D were determined. Results with the increase of exposure dose, there was a significant difference in renal development between cisplatin exposure group and Cisplatin exposure group (P0.01), and there was a dose-dependent (P0.01) RT-PCR result of Col _ 2AlPPA-Runx _ (2) and SOX9 MMP13GADPH, showing that there was no significant difference in renal development between cisplatin exposure group and Cisplatin exposure group (P0.01). The expression of osteogenesis related genes decreased (P0.05) and the ALP and TRAP of tissue sections decreased with the increase of cisplatin exposure dose (P0.05). Conclusion: it is feasible to establish chicken embryo kidney injury model by using cisplatin. 2) there is no direct effect of cisplatin on the bone development of chicken embryo. It mainly affects the function of osteoblasts and osteoclasts.
【学位授予单位】：暨南大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R692

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