背景：肝纤维化是一种癌前病变，受损肝细胞在纤维化微环境中转化为恶性病变的机制尚不明确。衰老是激活肝星状细胞（HSCs）的主要命运之一。关于衰老的HSCs对脂肪肝肝细胞行为的影响，目前文献资料不多。方法：使用小鼠非酒精性脂肪性肝炎（NASH）-纤维化-肝细胞癌（HCC）模型和人体NASH-HCC标本，鉴定衰老的HSCs。通过无标记质谱分析（NanoRPLC-MS/MS）和定量验证，分析衰老的HSCs的分泌物组成。结果：随着从非酒精性脂肪肝（NAFL）、NASH到NASH-纤维化的病情进展，衰老的HSCs逐渐增加，在晚期纤维化阶段达到峰值，随后在肝细胞畸形或HCC发展时下降。从衰老的HSCs的分泌物中鉴定出影响增殖、上皮间质转化（EMT）或迁移的关键成分，并可能激活形态发生孤立子或致癌Wnt信号通路，加速脂肪变性或畸形肝细胞的恶性转化。受刺激的原代肝细胞与衰老的HSCs的培养基共培养或在3D球体中共培养，显示出增强的增殖或EMT特性。结论：衰老的HSCs分泌一种特征蛋白质谱，有利于脂肪变性或畸形肝细胞通过激活形态发生孤立子或致癌Wnt信号通路，从NASH进展为恶性病变。©作者。

Background: Hepatic fibrosis is a premalignant lesion, and how injured hepatocytes transform into malignancy in a fibrotic microenvironment is poorly understood. Senescence is one of major fates of activated hepatic stellate cells (HSCs). Paucity of literature is available regarding the influence of senescent HSCs on behavior of steatotic hepatocytes. Methods: Senescent HSCs were identified in a murine model of nonalcoholic steatohepatitis (NASH)-fibrosis-hepatocellular carcinoma (HCC) and human NASH-HCC specimens. Secretome of senescent HSCs was analyzed by label-free mass-spectrum (NanoRPLC-MS/MS) and verified quantitatively. Results: Senescent HSCs were increased along with the progression from nonalcoholic fatty liver (NAFL), NASH to NASH-fibrosis, and reached a peak at the stage of advanced fibrosis and then decreased when hepatocellular dysplasia or HCC was developed. Critical components affecting proliferation, epithelial-mesenchymal transition (EMT) or migration were identified from secretome of senescent HSCs, and may activate morphogenic hedgehog or oncogenic Wnt signaling pathways to accelerate malignant transformation of steatotic or dysplastic hepatocytes. Primary hepatocytes stimulated with conditioned medium from senescent HSCs, in co-culture or co-cultured in 3D spheroids with senescent HSCs exhibited an enhanced proliferating or EMT profile. Conclusion: Senescent HSCs secreted a characterized protein profile favoring malignant transformation of steatotic or dysplastic hepatocytes through activating morphogenic hedgehog or oncogenic Wnt signaling pathways in the progression from NASH to malignancy.© The author(s).