细胞外基质纤维连接蛋白在小鼠肾脏发育中的表达*

王堃1，田鹤2，王岩2，郭敏2

课题背景：课题受辽宁省教育厅基金项目资助（05L134）。本课题组观察多种细胞外基质成分在正常小鼠肾脏发育过程中表达规律，为应用小鼠作为转基因动物模型的肾脏疾病研究提供基本的实验依据。�

应用要点：以往关于纤维连接蛋白在小鼠肾脏发育中的表达多局限于定性研究，本实验采用免疫组织化学技术、体视学方法和Western blot技术检测纤维连接蛋白在小鼠肾脏发育中的表达及其含量的变化，进行了系统的定性及半定量分析。实验结果表明，纤维连接蛋白对肾脏各结构的发育以及成熟肾脏各结构的维持都起到了重要作用。

术语解析：纤维连接蛋白是细胞外基质和血浆的一种主要成分，也是某些整合素黏附受体的特异性配体。纤维连接蛋白具有多方面生理功能 ,诸如:参与细胞与细胞、细胞与基质之间的粘连,维持细胞正常形态,保持微血管完整性,形成利于细胞运动的暂时性基质,促进细胞分化,参与伤口的愈合,阻止肿瘤细胞转移,提高网状内皮系统防御功能,调理单核巨噬细胞吞噬外来抗原,并能促进机体清除血中有害物质,包括免疫复合物、损伤的血小板、凝血产物、肿瘤细胞等。

摘要
目的:纤维连接蛋白作为细胞外基质的重要成份，成为肾脏正常组织结构的基本组成部分。实验观察纤维连接蛋白在小鼠肾脏发育中的时空性表达，探讨纤维连接蛋白与肾脏发育的关系。
方法：实验于2006-09/2007-07在辽宁医学院组织胚胎学教研室与科学实验中心完成。①实验材料：选取成年健康昆明系小白鼠44只，体质量25~30 g。②实验过程：小白鼠雌雄（1∶1）同窝饲养，以观察到阴道栓脱落的最早时间计为胚龄0 d,以观察到仔鼠出生的最早时间计为生后0 d。分别取胚龄12，14，16，18 d胎鼠和生后1，3，7，14，21，28，42 d仔鼠肾脏，每时间点4只，常规石蜡标本制备、切片。③实验评估：采用免疫组织化学方法、体视学和Western blot技术测定不同胚龄及生后日龄小鼠肾脏内纤维连接蛋白的表达及其含量变化。实验过程中对动物的处理符合动物伦理学标准。
结果：①免疫组织化学结果：显示从胚龄14 d开始，纤维连接蛋白在除逗号小体以外的各期肾小体、肾小管、集合管的基底膜处均有表达。②体视学测量结果：显示纤维连接蛋白在除逗号小体以外的各期肾小体及肾小管、集合管基底膜处的表达量均逐渐增加。③Western blot 结果：显示纤维连接蛋白随着肾脏的发育其表达量逐渐增加。
结论：纤维连接蛋白对肾脏各结构的发育以及成熟肾脏各结构的维持均起一定的作用。
关键词：纤维连接蛋白；肾脏；发育；小鼠组织构建

王堃，田鹤，王岩，郭敏.细胞外基质纤维连接蛋白在小鼠肾脏发育中的表达[J].中国组织工程研究与临床康复，2008，12(11):2087-2091 [www.zglckf.com/zglckf/ejournal/upfiles/08-11/11k-2087(ps).pdf]

王堃，男，1954年生，辽宁省锦西市人，汉族，1976年辽宁中医学院毕业，副主任医师，主要从事肾脏发生发育方面的研究。

通讯作者：郭敏，博士，教授，研究生导师，辽宁医学院组织胚胎学教研室，辽宁省锦州市 121001
mguo1118@yahoo.
com.cn

辽宁省教育厅基金项目资助（05L134）*

中图分类号:R329.461
文献标识码:B
文章编号:1673-8225
(2008)11-02087-05

收稿日期：2007-08-18
修回日期：2007-11-03
(07-50-8-4494/W·A）

Abstract

AIM：As an important part of extracellular matrix, fibronectin is the fundamental composition of normal kidney. In this study, we observed the chronological and spatial expression of fibronectin in mouse to investigate the correlation between fibronectin and kidney development.
METHODS: The experiment was performed in the Scientific Experiment Center and Department of Histology and Embryology, Liaoning Medical University from September 2006 to July 2007. ①Forty-four healthy adult Kunming mice weighing 25-30 g were selected and fed with the female and male at a ratio of 1∶1. The earliest time that vaginal suppository shed from the female mice was counted for embryonic day 0, and the earliest time that the neonatal mouse was born for postnatal day 0. The mice pregnant for 12，14, 16 and 18 days and mice born for 1, 3, 7, 14, 21, 28, and 42 days were selected with 4 mice at each time point. The kidneys were embedded in paraffin, and made into section. ②The expressions and content changes of fibronectin in embryos and postnatal mouse kidneys were examined by immunohistochemistry, stereological method and Western blot. All the procedure was accorded with animal ethical standards.
RESULTS: ①Immunohistochemical analysis showed that since pregnant for 12 days, fibronectin stained in the basal membrane of renal glomeruli, tubules and collecting duct in all developing stages except of the comma-shaped bodies. ②Stereological results suggested that the expressions of fibronectin were increased gradually in all stage of renal glomeruli and tubules, except of the comma-shaped bodies. ③Western blot results showed that the content of fibronectin was increased with kidney development.
CONCLUSION: Fibronectin plays an important role in kidney development and maturation.

Wang K, Tian H, Wang Y, Guo M.Fibronectin expression during mouse kidney development.Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu 2008;12(11):2087-2091(China) [www.zglckf.com/zglckf/ejournal/upfiles/08-11/11k-2087(ps).pdf]

1First Affiliated Hospital, 2Department of Histology and Embryology, Liaoning Medical College, Jinzhou 121001, Liaoning Province, China

Wang Kun, Associate chief physician, First Affiliated Hospital, Liaoning Medical College, Jinzhou 121001, Liaoning Province, China

Correspondence to: Guo Min, Doctor, Professor, Tutor of master, Department of Histology and Embryology, Liaoning Medical College, Jinzhou 121001, Liaoning Province, China
mguo1118@yahoo.
com.cn

Supported by: a grant from Education Department of Liaoning Province, No. 05L134*

Received: 2007-08-18
Accepted: 2007-11-03