Korovila Ioanna, Jung Tobias, Deubel Stefanie, Grune Tilman, Ott Christiane
Department of Molecular Toxicology, German Institute of Human Nutrition (DIfE) Potsdam-Rehbruecke, Nuthetal, 14558, Germany.
German Centre for Cardiovascular Research (DZHK), partner site Berlin, Berlin, 13347, Germany.
Mol Nutr Food Res. 2020 Oct;64(20):e2000816. doi: 10.1002/mnfr.202000816. Epub 2020 Sep 28.
Several studies show that excessive lipid intake can cause hepatic steatosis. To investigate lipotoxicity on cellular level, palmitate (PA) is often used to highly increase lipid droplets (LDs). One way to remove LDs is autophagy, while it is controversially discussed if autophagy is also affected by PA. It is aimed to investigate whether PA-induced LD accumulation can impair autophagy and punicalagin, a natural autophagy inducer from pomegranate, can improve it.
To verify the role of autophagy in LD degradation, HepG2 cells are treated with PA and analyzed for LD and perilipin 2 content in presence of autophagy inducer Torin 1 and inhibitor 3-Methyladenine. PA alone seems to initially induce autophagy-related proteins but impairs autophagic-flux in a time-dependent manner, considering 6 and 24 h PA. To examine whether punicalagin can prevent autophagy impairment, cells are cotreated for 24 h with PA and punicalagin. Results show that punicalagin preserves expression of autophagy-related proteins and autophagic flux, while simultaneously decreasing LDs and perilipin 2.
Data provide new insights into the role of PA-induced excessive LD content on autophagy and suggest autophagy-inducing properties of punicalagin, indicating that punicalagin can be a health-beneficial compound for future research on lipotoxicity in liver.
多项研究表明,脂质摄入过多会导致肝脂肪变性。为了在细胞水平上研究脂毒性,通常使用棕榈酸酯(PA)来大量增加脂滴(LDs)。清除脂滴的一种方法是自噬,然而自噬是否也受PA影响仍存在争议。本研究旨在探讨PA诱导的脂滴积累是否会损害自噬,以及石榴中的天然自噬诱导剂 punicalagin 是否能改善这种情况。
为了验证自噬在脂滴降解中的作用,用PA处理HepG2细胞,并在存在自噬诱导剂Torin 1和抑制剂3-甲基腺嘌呤的情况下分析脂滴和周脂素2的含量。单独使用PA似乎最初会诱导自噬相关蛋白,但会以时间依赖性方式损害自噬通量,以6小时和24小时的PA处理为例。为了研究punicalagin是否能预防自噬损伤,细胞用PA和punicalagin共同处理24小时。结果表明,punicalagin能保持自噬相关蛋白的表达和自噬通量,同时减少脂滴和周脂素2。
这些数据为PA诱导的过量脂滴含量对自噬的作用提供了新的见解,并表明punicalagin具有自噬诱导特性,这表明punicalagin可能是未来肝脏脂毒性研究中一种有益健康的化合物。