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S100B在小胶质细胞中的表达及其对小胶质细胞的影响。

S100B expression in and effects on microglia.

作者信息

Adami C, Sorci G, Blasi E, Agneletti A L, Bistoni F, Donato R

机构信息

Section of Microbiology, University of Perugia, Via del Giochetto, C.P. 81 Succ. 3, 06122 Perugia, Italy.

出版信息

Glia. 2001 Feb;33(2):131-42.

Abstract

We evaluated the intracellular and extracellular biological role of S100B protein with respect to microglia. S100B, which belongs to the multigenic family of Ca2+-binding proteins, is abundant in astrocytes where it is found diffusely in the cytoplasm and is associated with membranes and cytoskeleton constituents. S100B protein is also secreted by astrocytes and acts on these cells to stimulate nitric oxide secretion in an autocrine manner. However, little is known about the relationship between S100B and microglia. To address this issue, we used primary microglia from newborn rat cortex and the BV-2 microglial cell line, a well-established cell model for the study of microglial properties. S100B expression was assessed by immunofluorescence in primary microglia and by RT-PCR, Western blotting, and immunofluorescence in BV-2 cells. S100B was found in microglia in the form of a filamentous network as well as diffusely in the cytoplasm and associated with intracellular membranes. S100B relocated around phagosomes during BV-2 phagocytosis of opsonized Cryptococcus neoformans. Furthermore, interferon-gamma (IFN-gamma) treatment caused cell shape changes and redistribution of S100B, and downregulation of S100B mRNA expression in BV-2 cells. Treatment of BV-2 cells with nanomolar to micromolar amounts of S100B resulted in increased IFN-gamma-induced expression of inducible nitric oxide synthase mRNA as well as nitric oxide secretion. Taken together, these data suggest a possible role for S100B in the accomplishment/regulation of microglial cell functions.

摘要

我们评估了S100B蛋白在小胶质细胞方面的细胞内和细胞外生物学作用。S100B属于钙结合蛋白的多基因家族,在星形胶质细胞中含量丰富,在细胞质中呈弥散分布,并与膜和细胞骨架成分相关。S100B蛋白也由星形胶质细胞分泌,并以自分泌方式作用于这些细胞,刺激一氧化氮分泌。然而,关于S100B与小胶质细胞之间的关系知之甚少。为了解决这个问题,我们使用了新生大鼠皮质的原代小胶质细胞和BV-2小胶质细胞系,这是一个用于研究小胶质细胞特性的成熟细胞模型。通过原代小胶质细胞中的免疫荧光以及BV-2细胞中的RT-PCR、蛋白质印迹和免疫荧光来评估S100B的表达。在小胶质细胞中发现S100B以丝状网络的形式存在,也弥散于细胞质中并与细胞内膜相关。在BV-2吞噬调理过的新型隐球菌期间,S100B重新定位到吞噬体周围。此外,干扰素-γ(IFN-γ)处理导致细胞形态改变和S100B重新分布,并下调BV-2细胞中S100B mRNA的表达。用纳摩尔至微摩尔量的S100B处理BV-2细胞导致IFN-γ诱导的诱导型一氧化氮合酶mRNA表达以及一氧化氮分泌增加。综上所述,这些数据表明S100B在小胶质细胞功能的完成/调节中可能发挥作用。

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