Abstract

Microglia phagocytosis plays an important role in the pathogenesis of neurodegeneration. Defects or dysfunction of microglia phagocytosis were observed in neurodegenerative diseases, with different targets and associated receptors influencing the microglia response. Moreover, non-canonical LC3-associated (microtubule-associated protein 1 light chain 3) phagocytosis was extensively studied recently as a novel form of phagocytosis on macrophages, but little on microglia. Here, we investigated changes in phagocytic function of microglia activated by lipopolysaccharide (LPS) as well as rapamycin-regulated phagocytosis. Phagocytosis in mouse BV2 cells and primary microglia was analyzed by flow cytometry and immunofluorescence. Phagocytosis-related mechanisms in BV2 cells were analyzed using Western blotting and real-time polymerase chain reaction. Rapamycin was shown to reduce LPS-induced phagocytosis of microglia and at the same time stimulate LС3-dependent phagocytosis by regulating Atg3, Atg4 and Atg7 expression. In addition, in BV2 cells, the PI3K/AKT/mTOR pathway may be involved in phagocytosis. These results suggest that phagocytosis of microglia is a complex process, and the increase in phagocytosis should not be considered as a maturation of phagocytic function. The data will provide new insights into the mechanisms of phagocytosis and neuroimmunity.

中文翻译：

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雷帕霉素在体外调节脂多糖诱导的小胶质细胞吞噬作用

摘要

小胶质细胞吞噬作用在神经退行性变的发病机制中发挥着重要作用。在神经退行性疾病中观察到小胶质细胞吞噬作用的缺陷或功能障碍，不同的靶标和相关受体影响小胶质细胞的反应。此外，非经典 LC3 相关（微管相关蛋白 1 轻链 3）吞噬作用最近作为巨噬细胞吞噬作用的一种新型形式得到了广泛研究，但对小胶质细胞的吞噬作用却很少。在这里，我们研究了脂多糖（LPS）激活的小胶质细胞吞噬功能的变化以及雷帕霉素调节的吞噬作用。通过流式细胞术和免疫荧光分析小鼠 BV2 细胞和原代小胶质细胞的吞噬作用。使用蛋白质印迹和实时聚合酶链反应分析 BV2 细胞中吞噬作用相关的机制。Rapamycin 可减少 LPS 诱导的小胶质细胞吞噬作用，同时通过调节Atg3、Atg4和Atg7的表达来刺激 LС3 依赖性吞噬作用。此外，在BV2细胞中，PI3K/AKT/mTOR通路可能参与吞噬作用。这些结果表明小胶质细胞的吞噬作用是一个复杂的过程，吞噬作用的增加不应被视为吞噬功能的成熟。这些数据将为吞噬作用和神经免疫机制提供新的见解。