Goeritzer Madeleine, Vujic Nemanja, Schlager Stefanie, Chandak Prakash G, Korbelius Melanie, Gottschalk Benjamin, Leopold Christina, Obrowsky Sascha, Rainer Silvia, Doddapattar Prakash, Aflaki Elma, Wegscheider Martin, Sachdev Vinay, Graier Wolfgang F, Kolb Dagmar, Radovic Branislav, Kratky Dagmar
Institute of Molecular Biology & Biochemistry, Center of Molecular Medicine, Medical University of Graz, Harrachgasse 21, 8010 Graz, Austria.
Center for Medical Research/Institute of Cell Biology, Histology and Embryology, Medical University of Graz, Harrachgasse 21, 8010 Graz, Austria.
Biochim Biophys Acta. 2015 Oct;1851(10):1304-1316. doi: 10.1016/j.bbalip.2015.06.005. Epub 2015 Jul 2.
During autophagy, autophagosomes fuse with lysosomes to degrade damaged organelles and misfolded proteins. Breakdown products are released into the cytosol and contribute to energy and metabolic building block supply, especially during starvation. Lipophagy has been defined as the autophagy-mediated degradation of lipid droplets (LDs) by lysosomal acid lipase. Adipose triglyceride lipase (ATGL) is the major enzyme catalyzing the initial step of lipolysis by hydrolyzing triglycerides (TGs) in cytosolic LDs. Consequently, most organs and cells, including macrophages, lacking ATGL accumulate TGs, resulting in reduced intracellular free fatty acid concentrations. Macrophages deficient in hormone-sensitive lipase (H0) lack TG accumulation albeit reduced in vitro TG hydrolase activity. We hypothesized that autophagy is activated in lipase-deficient macrophages to counteract their energy deficit. We therefore generated mice lacking both ATGL and HSL (A0H0). Macrophages from A0H0 mice showed 73% reduced neutral TG hydrolase activity, resulting in TG-rich LD accumulation. Increased expression of cathepsin B, accumulation of LC3-II, reduced expression of p62 and increased DQ-BSA dequenching suggest intact autophagy and functional lysosomes in A0H0 macrophages. Markedly decreased acid TG hydrolase activity and lipid flux independent of bafilomycin A1 treatment, however, argue against effective lysosomal degradation of LDs in A0H0 macrophages. We conclude that autophagy of proteins and cell organelles but not of LDs is active as a compensatory mechanism to circumvent and balance the reduced availability of energy substrates in A0H0 macrophages.
在自噬过程中,自噬体与溶酶体融合以降解受损的细胞器和错误折叠的蛋白质。分解产物被释放到细胞质中,为能量和代谢构建块的供应做出贡献,尤其是在饥饿期间。脂肪自噬被定义为溶酶体酸性脂肪酶介导的脂滴(LDs)的自噬性降解。脂肪甘油三酯脂肪酶(ATGL)是催化脂解第一步的主要酶,通过水解细胞质中脂滴中的甘油三酯(TGs)来实现。因此,大多数器官和细胞,包括巨噬细胞,缺乏ATGL会积累TGs,导致细胞内游离脂肪酸浓度降低。缺乏激素敏感性脂肪酶(H0)的巨噬细胞尽管体外TG水解酶活性降低,但缺乏TG积累。我们假设自噬在脂肪酶缺陷的巨噬细胞中被激活,以抵消其能量不足。因此,我们培育了同时缺乏ATGL和HSL(A0H0)的小鼠。来自A0H0小鼠的巨噬细胞显示中性TG水解酶活性降低了73%,导致富含TG的脂滴积累。组织蛋白酶B表达增加、LC3-II积累、p62表达降低以及DQ-BSA去淬灭增加表明A0H0巨噬细胞中自噬完整且溶酶体功能正常。然而,与巴弗洛霉素A1处理无关的酸性TG水解酶活性和脂质通量显著降低,这表明A0H0巨噬细胞中脂滴的溶酶体有效降解存在问题。我们得出结论,蛋白质和细胞器的自噬而非脂滴的自噬作为一种补偿机制是活跃的,以规避和平衡A0H0巨噬细胞中能量底物可用性的降低。