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斜纹枯叶蛾刚毛提取物调节Rac1激活、膜动力学和细胞黏附特性。

Lonomia obliqua bristle extract modulates Rac1 activation, membrane dynamics and cell adhesion properties.

作者信息

Bernardi L, Pinto A F M, Mendes E, Yates J R, Lamers M L

机构信息

Basic Research Center, Dentistry School, Federal University of Rio Grande of Sul, Brazil; Department of Morphological Sciences, Institute of Basic Health Sciences, Federal University of Rio Grande do Sul, Brazil.

Clayton Foundation Peptide Biology Lab, Salk Institute for Biological Studies, USA.

出版信息

Toxicon. 2019 Apr 15;162:32-39. doi: 10.1016/j.toxicon.2019.02.019. Epub 2019 Mar 6.

Abstract

Lonomia obliqua is a caterpillar of potential therapeutic interest whose venom is able to induce severe blood leakage and modulate leukocyte migration. Since both phenotypes are associated with changes in cytoskeleton dynamics and cell adhesion properties, the aim of this study was to analyze the effects of Lonomia obliqua bristle extract (LOBE) in cell adhesion and migration signaling. Proteomic analysis revealed that epithelial cells (CHO-K1) exposed to LOBE (30 μg/mL, 30 min) exhibited changes in levels of actin regulatory proteins, including RhoGTPases. These changes correlated with an increase in the activity of the RhoGTPase family member Rac as measured by Förster resonance energy transfer (FRET). When plated in migration promoting conditions, CHO-K1 cells exposed to LOBE (10 μg/mL) showed an increase in membrane ruffling after short (30 min) period of incubation that was accompanied by changes in the distribution of the adhesion markers paxillin, vinculin and an increase of focal adhesion kinase autophosphorylation levels (Y397), suggesting changes in cell-extracellular matrix (ECM) adhesion properties and signaling. These data suggest that LOBE possesses bioactive molecules that are capable to modulated cell migration signaling, cytoskeletal dynamics and cell-ECM properties of several cell types.

摘要

斜纹脂毛虫是一种具有潜在治疗价值的毛虫,其毒液能够引发严重的血液渗漏并调节白细胞迁移。由于这两种表型都与细胞骨架动力学和细胞黏附特性的变化有关,本研究的目的是分析斜纹脂毛虫刚毛提取物(LOBE)对细胞黏附及迁移信号传导的影响。蛋白质组学分析显示,暴露于LOBE(30μg/mL,30分钟)的上皮细胞(CHO-K1)中,包括RhoGTPases在内的肌动蛋白调节蛋白水平发生了变化。这些变化与通过荧光共振能量转移(FRET)测量的RhoGTPase家族成员Rac活性增加相关。当在促进迁移的条件下铺板时,暴露于LOBE(10μg/mL)的CHO-K1细胞在短时间(30分钟)孵育后膜褶皱增加,同时黏附标记桩蛋白、纽蛋白的分布发生变化,粘着斑激酶自磷酸化水平(Y397)升高,表明细胞与细胞外基质(ECM)的黏附特性和信号传导发生了变化。这些数据表明,LOBE含有能够调节几种细胞类型的细胞迁移信号传导、细胞骨架动力学和细胞与ECM特性的生物活性分子。

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