Novel defatting strategies reduce lipid accumulation in primary human culture models of liver steatosis
Novel defatting strategies reduce lipid accumulation in primary human culture models of liver steatosis
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DOI：
10.1242/dmm.042663
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发表时间：
2020-04-01
影响因子：
4.3
通讯作者：
Conti, Filomena
中科院分区：
文献类型：
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作者：
Aoudjehane, Lynda;Gautheron, Jeremie;Conti, Filomena
常温灌注提供了一种挽救脂肪变性肝移植物的方法，包括药物脱脂。在这项研究中，我们测试了新的药物组合在三种人类培养模型中触发细节的潜力，诱导脂肪变性的原代肝细胞，从脂肪变性肝脏分离的原代肝细胞，以及脂肪变性肝脏的精密切割肝脏切片（PCLS）。毛喉素、L-卡米汀和一种PPARalpha激动剂都与雷帕霉素（一种诱导自噬的免疫抑制剂）在D-FAT鸡尾酒中组合。D-FAT单独或与坏死磺酰胺（一种参与坏死性凋亡的混合谱系激酶结构域样假激酶的抑制剂）组合进行测试。在24小时内，在所有三个模型中。D-FAT诱导甘油三酯含量降低30%，这归因于参与游离脂肪酸β-氧化和自噬的基因的上调，以及参与脂肪生成的基因的下调。脱脂伴随着内质网应激和活性氧的产生减少。在PCLS中，添加坏死磺酰胺使脱脂功效增加8%-12%，具有增加自噬的趋势。总之，培养模型，特别是PCLS，对设计肝移植抢救策略很有见地。通过靶向线粒体氧化代谢、大细胞自噬和脂肪生成的药物组合可以快速实现脱脂。
Normotherrnic perfusion provides a means to rescue steatotic liver grafts, including by pharmacological defatting. In this study, we tested the potential of new drug combinations to trigger detailing in three human culture models, primary hepatocytes with induced steatosis, primary hepatocytes isolated from steatotic liver, and precision-cut liver slices (PCLS) of steatotic liver. Forskolin, L-camitine and a PPAR alpha agonist were all combined with rapamycin, an immunosuppressant that induces autophagy, in a D-FAT cocktail. D-FAT was tested alone or in combination with necrosulfonamide, an inhibitor of mixed lineage kinase domain like pseudokinase involved in necroptosis. Within 24 h, in all three models. D-FAT induced a decrease in triglyceride content by 30%, attributable to an upregulation of genes involved in free fatty acid beta-oxidation and autophagy, and a downregulation of those involved in lipogenesis. Defatting was accompanied by a decrease in endoplasmic reticulum stress and in the production of reactive oxygen species. The addition of necrosulfonamide increased the efficacy of defatting by 8%-12% in PCLS, with a trend towards increased autophagy. In conclusion, culture models, notably PCLS, are insightful to design strategies for liver graft rescue. Defatting can be rapidly achieved by combinations of drugs targeting mitochondrial oxidative metabolism, macro-autophagy and lipogenesis.