Miltefosine increases macrophage cholesterol release and inhibits NLRP3-inflammasome assembly and IL-1β release
Miltefosine increases macrophage cholesterol release and inhibits NLRP3-inflammasome assembly and IL-1β release
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DOI：
10.1038/s41598-019-47610-w
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发表时间：
2019-07-31
影响因子：
4.6
通讯作者：
Gulshan, Kailash
中科院分区：
文献类型：
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作者：
Iacano, Amanda J.;Lewis, Harvey;Gulshan, Kailash
米替福新是FDA批准的用于治疗皮肤和内脏利什曼病的口服药物。利什曼原虫是一种有鞭毛的原生动物，它感染巨噬细胞并在巨噬细胞中分化。在这里，我们研究了米替福新对巨噬细胞的脂质稳态、自噬和NLRP 3炎性小体组装/活性的影响。米替福新治疗对巨噬细胞脂质稳态产生多重影响，导致细胞胆固醇释放增加、脂筏破坏增加、细胞表面磷脂酰丝氨酸（PS）翻转减少以及磷脂酰肌醇4，5-二磷酸（PIP 2）从质膜重新分布至细胞中富含肌动蛋白的区域。与对照相比，在用米替福新处理的细胞中观察到增强的基础自噬、脂肪吞噬和线粒体自噬。米替福新处理的细胞显示出参与自噬诱导的激酶的磷酸化显著增加，所述激酶例如腺苷单磷酸活化蛋白激酶（AMPK）和Unc-51样自噬活化激酶（ULK 1）。Toll样受体（TLR）信号通路被米替福新处理钝化，导致TLR 4向细胞表面的募集减少，并且LPS诱导的pro-IL-1 β mRNA水平类似于75%的降低。米替福新可降低内毒素介导的线粒体活性氧，保护线粒体膜电位。米替福新治疗诱导线粒体自噬并抑制NLRP 3炎性体组装。总的来说，我们的数据显示米替福新诱导ABCA 1介导的胆固醇释放，诱导AMPK磷酸化和线粒体自噬，同时抑制NLRP 3炎性小体组装和IL-1 β释放。
Miltefosine is an FDA approved oral drug for treating cutaneous and visceral leishmaniasis. Leishmania is a flagellated protozoa, which infects and differentiates in macrophages. Here, we studied the effects of Miltefosine on macrophage's lipid homeostasis, autophagy, and NLRP3 inflammasome assembly/activity. Miltefosine treatment conferred multiple effects on macrophage lipid homeostasis leading to increased cholesterol release from cells, increased lipid-raft disruption, decreased phosphatidylserine (PS) flip from the cell-surface, and redistribution of phosphatidylinositol 4,5-bisphosphate (PIP2) from the plasma membrane to actin rich regions in the cells. Enhanced basal autophagy, lipophagy and mitophagy was observed in cells treated with Miltefosine vs. control. Miltefosine treated cells showed marked increased in phosphorylation of kinases involved in autophagy induction such as; Adenosine monophosphate-activated protein kinase (AMPK) and Unc-51 like autophagy activating kinase (ULK1). The Toll like receptor (TLR) signaling pathway was blunted by Miltefosine treatment, resulting in decreased TLR4 recruitment to cell-surface and similar to 75% reduction in LPS induced pro-IL-1 beta mRNA levels. Miltefosine reduced endotoxin-mediated mitochondrial reactive oxygen species and protected the mitochondrial membrane potential. Miltefosine treatment induced mitophagy and dampened NLRP3 inflammasome assembly. Collectively, our data shows that Miltefosine induced ABCA1 mediated cholesterol release, induced AMPK phosphorylation and mitophagy, while dampening NLRP3 inflammasome assembly and IL-1 beta release.