Department of Cardiovascular and Metabolic Sciences, Cleveland Clinic, Cleveland, OH, 44195, USA.
Department of Cardiovascular Medicine, Cleveland Clinic, Cleveland, OH, 44195, USA.
Sci Rep. 2019 Jul 31;9(1):11128. doi: 10.1038/s41598-019-47610-w.
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 ~75% reduction in LPS induced pro-IL-1β 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-1β release.
米替福新是一种经美国食品药品监督管理局批准的用于治疗皮肤利什曼病和内脏利什曼病的口服药物。利什曼原虫是一种鞭毛原生动物，它在巨噬细胞中感染和分化。在这里，我们研究了米替福新对巨噬细胞脂质稳态、自噬和 NLRP3 炎性体组装/活性的影响。米替福新处理对巨噬细胞脂质稳态产生了多种影响，导致细胞内胆固醇释放增加，脂筏破坏增加，细胞膜表面磷脂酰丝氨酸（PS）翻转减少，以及磷脂酰肌醇 4,5-二磷酸（PIP2）从质膜向细胞中富含肌动蛋白的区域重新分布。与对照组相比，用米替福新处理的细胞中观察到基础自噬、脂自噬和线粒体自噬增强。米替福新处理的细胞显示参与自噬诱导的激酶如腺苷单磷酸激活蛋白激酶（AMPK）和非典型卷曲相关激酶 1（ULK1）的磷酸化明显增加。米替福新处理阻断 Toll 样受体（TLR）信号通路，导致 TLR4 向细胞表面募集减少，LPS 诱导的 pro-IL-1β mRNA 水平降低约 75%。米替福新减少内毒素介导的线粒体活性氧并保护线粒体膜电位。米替福新处理诱导线粒体自噬并抑制 NLRP3 炎性体组装。总之，我们的数据表明，米替福新诱导 ABCA1 介导的胆固醇释放，诱导 AMPK 磷酸化和线粒体自噬，同时抑制 NLRP3 炎性体组装和 IL-1β 释放。
