Impact of inhibition of the autophagy-lysosomal pathway on biomolecules carbonylation and proteome regulation in rat cardiac cells
Impact of inhibition of the autophagy-lysosomal pathway on biomolecules carbonylation and proteome regulation in rat cardiac cells
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DOI：
10.1016/j.redox.2019.101123
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发表时间：
2019-05-01
期刊：
影响因子：
11.4
通讯作者：
Fedorova, Maria
中科院分区：
文献类型：
--
作者：
Coliva, Giulia;Duarte, Sofia;Fedorova, Maria
细胞采用多种防御机制来维持与修饰的自身生物分子的积累相关的广泛的应激条件，从而导致脂毒性和蛋白毒性。其中一种机制涉及自噬-溶酶体途径的激活，以去除和降解修饰的脂质、蛋白质甚至细胞器。生物分子羰基化是一种不可逆的氧化修饰，在多种病理条件下发生，通常被视为氧化应激的标志。在这里，我们使用大鼠原代心肌细胞模型来阐明自噬-溶酶体途径在羰基化生物分子周转中的作用。用荧光显微镜研究了用自噬-溶酶体降解抑制剂处理的细胞，并用短脉冲的轻度氮氧化应激处理，观察到羰基化生物分子在液滴或囊泡样结构中的积累。此外，使用相对无标记定量方法的蛋白质组调控的全系统分析揭示了用蛋白酶抑制剂处理的细胞中最显著的改变。有趣的是，胰岛素信号转导的下调是最丰富的途径之一，如通过调节蛋白质的功能注释所揭示的。
Cells employ multiple defence mechanisms to sustain a wide range of stress conditions associated with accumulation of modified self-biomolecules leading to lipo- and proteotoxicity. One of such mechanisms involves activation of the autophagy-lysosomal pathway for removal and degradation of modified lipids, proteins and even organelles. Biomolecules carbonylation, an irreversible oxidative modification, occurs in a variety of pathological conditions and is generally viewed as a marker of oxidative stress. Here, we used a model of rat primary cardiac cells to elucidate the role of autophagy-lysosomal pathway in the turnover of carbonylated biomolecules. Cells treated with inhibitors of autophagy-lysosomal degradation and primed with a short pulse of mild nitroxidative stress were studied using fluorescent microscopy and accumulation of carbonylated biomolecules in droplets- or vesicle-like structures was observed. Furthermore, systems-wide analysis of proteome regulation using relative label free quantification approach revealed the most significant alterations in cells treated with protease inhibitors. Interestingly, down-regulation of insulin signalling was among the most enriched pathway, as revealed by functional annotation of regulated proteins.