巢蛋白对维持足细胞形态作用的探讨  

Role of nestin in the maintenance of the structure of podocytes

作　　者：苏蔚[1,2] 房成[2] 杨海春[1] 陈靖[1] 顾勇[1] 郝传明[1] 

机构地区：[1]复旦大学附属华山医院肾脏科,上海200040 [2]新乡医学院病理教研室,河南新乡453003

出　　处：《中国病理生理杂志》2009年第1期156-159,共4页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30570861)

摘　　要：目的:观察细胞骨架蛋白巢蛋白(nestin)在体外培养的小鼠足细胞中的表达及在维持足细胞形态方面的作用。方法:应用免疫荧光技术检测nestin在条件性永生的足细胞中的表达;应用RNA干扰技术抑制巢蛋白nestin在足细胞中的表达,观察足细胞形态的变化并计算nestin siRNA组和无关序列siRNA组中有足突的足细胞数量。结果:体外培养的足细胞中巢蛋白呈丝状分布于胞浆;转染nestin siRNA质粒的足细胞,足突变短或消失,而转染无关对照序列siRNA质粒的足细胞形态未见改变;无关对照组有足突的足细胞数为77.0%±6.3%,转染nestin siRNA序列质粒组有足突的细胞数为16.0%±4.6%,两组差异显著(n=3,P<0.01)。结论:巢蛋白nes-tin对于维持足细胞正常形态具有重要作用。AIM： To investigate the expression of nestin, a kind of cytoskeletal protein in cultured murine podocytes and the role of nestin in the maintenance of the podocyte structure. METHODS ： The immortalized murine podocytes were cultured. The expression of nestin was determined by immunofluorescence. In differentiated podocytes, the expression of nestin was knock - down by RNAi. The effect of nestin knock - down was examined by Western blotting and immunofluorescence. The projections longer than maximal length of the cell body in which the cells transfected with siRNA and control vector that contained nonhomologous oligo were counted, respectively. RESULTS： Nestin siRNA markedly reduced or abolished nestin expression. In ceils transfected with nestin siRNA, the percentage of cells with processes was significantly lower than that in cells transfected with control vector （77.0% ± 6. 3% vs 16.0% ±4. 6%, n = 3, P 〈 0. 01 ）. CONCLUSION： Nestin may play an important role in maintaining normal function of podocytes.

关 键 词：足细胞 巢蛋白 RNA干扰 免疫荧光 

分 类 号：R3[医药卫生—基础医学]