足细胞中巢蛋白的表达及其相互作用分子的研究

目的 观察细胞骨架蛋白巢蛋白(Nestin)在肾脏足细胞的表达及其在足细胞中的作用方式。方法 免疫组化、免疫荧光、免疫电镜方法检测Nestin在人正常肾组织、体外培养的小鼠足细胞中的表达。免疫共沉淀方法检测足细胞中与巢蛋白相互作用的蛋白分子。结果 在正常肾组织中巢蛋白的表达局限于肾小球。免疫电镜证实巢蛋白主要表达于足细胞胞浆及初级足突，体外培养的足细胞中也可观察到巢蛋白丝状分布于胞浆。免疫共沉淀显示足细胞中Nestin与另两种细胞骨架蛋白波形蛋白(vimentin)、α-internexin相互作用。 结论 巢蛋白除了表达于中枢神经干细胞、胰岛干细胞等之外，在肾脏足细胞中也有表达，并与足细胞中其他细胞骨架蛋白相互作用，共同维持其正常形态和功能。

The research of Nestin expression and the interacted proteins in human podocytes

Objective To investigate the expression of Nestin， a kind of cytoskeletal protein，in human podocytes. Methods The expression of Nestin in normal human kidney and cultured murine podocytes was determined by immunoelectromicroscope， immunohistochemistry， and immunofluorescence. Co-immunopricipitation (co-IP) was used to detect the proteins that interacted with nestin in podocytes. Results Nestin was selectively expressed in human podocytes and localized in the primary and second processes. The co-immunopricipitation studies demonstrated that Nestin had a physical interaction with vimentin and α-internexin， but not with alpha-actinin 4. Conclusions Nestin is selectively expressed in glomerular podocytes of human kidney，central nerve stem cells，and pancreatic island stem cells as well. Nestin can interact with other cytoskeletal protein in podocyte. This study suggests the potential role of nestin in modulating normal cytoskeletal structure of podocytes.