自噬在高糖诱导的巨噬细胞经典活化中的作用
首发时间:2018-05-10
摘要:目的:巨噬细胞浸润是糖尿病肾病的重要组织病理特征,探讨自噬对高糖环境下巨噬细胞活化状态的影响,并进一步探索自噬流各阶段对巨噬细胞 M1/M2 表型转化的影响。 方法:体外培养小鼠巨噬细胞系 RAW264.7,通过 Western blot、免疫荧光检测 30mmol/L 高糖环境下巨噬细胞表型及自噬相关蛋白标志物的表达情况。利用雷帕霉素(促进自噬体生成)、3-甲基腺嘌呤(抑制自噬体生成)、巴弗洛霉素A1(抑制自噬体与溶酶体融合)及氯喹(抑制自噬溶酶体降解) 干预 M1/M2 经典模型中的自噬流各阶段,评估对巨噬细胞表型及自噬标志物的影响。 结果:(1)高糖诱导 RAW264.7 细胞 M1 型活化,同时抑制 LC3、Beclin-1 表达,促进 p62 的表达,降低了其自噬水平;提高自噬水平时则趋向于 M2 型活化。(2)RAW264.7 细胞的经典 M1 模型的自噬水平显著高于 M2 型,同时电镜观察到其自噬流过程。(3)分别干预自噬流各阶段,其自噬相关标志物的变化趋势明显不同,但均会影响 M1/M2 型相互转化,即激活自噬可诱导细胞向 M2 型转化,而抑制自噬则诱导 M1 型活化。 结论:高糖诱导巨噬细胞 M1 型活化,与其自噬降低有关,自噬流各阶段均能调节巨噬细胞 M1/M2 表型相互转化。单一标志物的变化不能判定细胞的自噬活性变化,需要评估自噬流的动态变化。
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The Effects of Autophagy on Classical Activation of Macrophages Induced by High Glucose
Abstract:Objective:Macrophage infiltration is an important histopathological feature of diabetic nephropathy. To investigate the effect of autophagy on macrophage activation in high glucose environment, and to further explore the effect of autophagic flow on the macrophage M1 / M2 phenotype transition. Methods: The mouse macrophage RAW264.7 was cultured in vitro. The macrophage phenotype and autophagy-related protein marker expression were detected by Western blot aThe Effects of Autophagy on Classical Activation of Macrophages Induced by High Glucosend immunofluorescence at a high glucose concentration of 30 mmol/L. Use of rapamycin (promotes autophagosome production), 3-methyladenine (inhibits autophagosome production), bafilomycin A1 (inhibits autophagosome-lyase fusion), and chloroquine (inhibits autophagy) Lysosome Degradation) Interventions in various stages of autophagic flow in the classical M1/M2 model assess the effect on macrophage phenotype and autophagy markers. Results: (1) High glucose induces M1 activation in RAW264.7 cells, inhibits the expression of LC3 and Beclin-1, promotes the expression of p62, and decreases the level of autophagy; when the level of autophagy is increased, it tends to be activated by M2.(2) The autophagy level of the classic M1 model of RAW264.7 cells was significantly higher than that of the M2 type, and the autophagic flow process was observed by electron microscopy. (3) Interventions in the autophagic flow at different stages have significantly different trends in autophagy-related markers, but all affect the M1/M2 type transformation. That is, activation of autophagy induces M2-type transformation of the cells, while inhibition of autophagy induces M1-type activation. Conclusion: High glucose induces M1 activation in macrophages and is associated with decreased autophagy. Autophagic flow regulates the M1/M2 phenotype transition of macrophages at various stages. Changes in a single marker cannot be used to determine changes in the autophagic activity of cells, and the dynamic changes in autophagic flow need to be assessed.
Keywords: Macrophages Phenotypic transformation High glucose Autophagic flow
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