Non-alcoholic steatohepatitis (NASH) is a highly prevalent, chronic liver disease characterized by hepatic lipid accumulation, inflammation, and concomitant fibrosis. Up to date, no anti-NASH drugs have been approved. In this study, we reproduced key NASH characteristics in vitro by exposing primary human hepatocytes (PHH), human skin stem cell-derived hepatic cells (hSKP-HPC), HepaRG and HepG2 cell lines, as well as LX-2 cells to multiple factors that play a role in the onset of NASH. The obtained in vitro disease models showed intracellular lipid accumulation, secretion of inflammatory chemokines, induced ATP content, apoptosis, and increased pro-fibrotic gene expression. These cell systems were then used to evaluate the anti-NASH properties of eight peroxisome proliferator-activated receptor (PPAR) agonists (bezafibrate, elafibranor, fenofibrate, lanifibranor, pemafibrate, pioglitazone, rosiglitazone, and saroglitazar). PPAR agonists differently attenuated lipid accumulation, inflammatory chemokine secretion, and pro-fibrotic gene expression.

Based on the obtained readouts, a scoring system was developed to grade the anti-NASH potencies. The in vitro scoring system, based on a battery of the most performant models, namely PHH, hSKP-HPC, and LX-2 cultures, showed that elafibranor, followed by saroglitazar and pioglitazone, induced the strongest anti-NASH effects. These data corroborate available clinical data and show the relevance of these in vitro models for the preclinical investigation of anti-NASH compounds.

非酒精性脂肪性肝炎 (NASH) 是一种高度流行的慢性肝病，其特征是肝脏脂质积聚、炎症和伴随的纤维化。迄今为止，尚未批准任何抗 NASH 药物。在这项研究中，我们通过将原代人肝细胞 (PHH)、人皮肤干细胞衍生的肝细胞 (hSKP-HPC)、HepaRG 和 HepG2 细胞系以及 LX-2 细胞暴露于多种因素，在体外重现了关键的 NASH 特征这在 NASH 的发作中发挥作用。获得的体外疾病模型显示细胞内脂质积累、炎症趋化因子的分泌、诱导的 ATP 含量、细胞凋亡和促纤维化基因表达增加。然后使用这些细胞系统来评估八种过氧化物酶体增殖物激活受体 (PPAR) 激动剂（苯扎贝特、elafibranor、非诺贝特、lanifibranor、pemafibrate、吡格列酮、罗格列酮和沙罗格列扎）。PPAR 激动剂不同程度地减弱脂质积累、炎症趋化因子分泌和促纤维化基因表达。

基于获得的读数，开发了一个评分系统来对抗 NASH 效力进行分级。体外评分系统基于一组性能最佳的模型，即 PHH、hSKP-HPC 和 LX-2 培养物，表明 elafibranor，其次是 saroglitazar 和吡格列酮，诱导了最强的抗 NASH 作用。这些数据证实了可用的临床数据，并显示了这些体外模型与抗 NASH 化合物的临床前研究的相关性。