Lu Mengxin, Tao Shuai, Zhao Conglin, Wang Neng, Hu Qiankun, Li Qiang, Qi Xun, Li Xinyan, Zhang Yi, Tu Chuantao, Huang Yuxian, Chen Liang
Department of Liver Disease, Shanghai Public Health Clinical Center, Fudan University, No. 2901 Caolang Road, Jinshan District, Shanghai, China.
Scientific Research Center, Shanghai Public Health Clinical Center, Fudan University, Shanghai, China.
Cell Mol Life Sci. 2025 Apr 17;82(1):161. doi: 10.1007/s00018-025-05682-0.
Latent Transforming Growth Factor Beta Binding Protein 2 (LTBP2) is a multi-domain exocrine protein located in the extracellular matrix (ECM) and has been implicated in fibrosis across various organs. However, its role in liver fibrosis remains inadequately understood. This study aims to elucidate the function and mechanism of LTBP2 in hepatic stellate cells (HSCs) activation and liver fibrosis. Our findings indicate that LTBP2 expression is positively correlated with liver fibrosis and is significantly elevated in fibrotic liver tissues from both human and murine models. Importantly, AAV6-mediated knockdown of LTBP2 in HSCs markedly alleviates CCl4-induced liver fibrosis by inhibiting the HSCs activation and reducing collagen deposition in mice. Gain-of-function and loss-of-function experiments confirmed that overexpression or knockdown of LTBP2 can enhance or inhibit the activation of HSCs, proliferation, migration and epithelial-mesenchymal transition (EMT) in LX-2 cells. Mechanistically, chromatin immunoprecipitation (ChIP) assays and dual-luciferase reporter gene assays revealed that Hypoxia-inducible Factor 1α (HIF-1α) promotes LTBP2 expression by directly binding to the LTBP2 promoter region. Furthermore, molecular docking and co-immunoprecipitation (Co-IP) experiments demonstrated an interaction between Lysyl Oxidase Like Protein 1 (LOXL1) and LTBP2. Rescue experiments verified that LTBP2 interacts with LOXL1 via the ERK signaling pathway to promote the activation of HSCs and EMT. Our results provide compelling evidence that the HIF-1α/LTBP2 axis facilitates the activation of HSCs and EMT by interacting with LOXL1 through ERK signaling pathway, suggesting that LTBP2 may serve as a potential therapeutic target for liver fibrosis.
潜伏转化生长因子β结合蛋白2(LTBP2)是一种位于细胞外基质(ECM)中的多结构域外分泌蛋白,与多种器官的纤维化有关。然而,其在肝纤维化中的作用仍未得到充分了解。本研究旨在阐明LTBP2在肝星状细胞(HSCs)激活和肝纤维化中的功能及机制。我们的研究结果表明,LTBP2表达与肝纤维化呈正相关,在人类和小鼠模型的纤维化肝组织中均显著升高。重要的是,腺相关病毒6(AAV6)介导的HSCs中LTBP2基因敲低通过抑制HSCs激活和减少小鼠体内胶原蛋白沉积,显著减轻了四氯化碳诱导的肝纤维化。功能获得和功能丧失实验证实,LTBP2的过表达或敲低可增强或抑制LX-2细胞中HSCs的激活、增殖、迁移和上皮-间质转化(EMT)。机制上,染色质免疫沉淀(ChIP)实验和双荧光素酶报告基因实验表明,缺氧诱导因子1α(HIF-1α)通过直接结合LTBP2启动子区域促进LTBP2表达。此外,分子对接和免疫共沉淀(Co-IP)实验证明了赖氨氧化酶样蛋白1(LOXL1)与LTBP2之间存在相互作用。挽救实验证实,LTBP2通过ERK信号通路与LOXL1相互作用,促进HSCs激活和EMT。我们的结果提供了有力证据,即HIF-1α/LTBP2轴通过ERK信号通路与LOXL1相互作用促进HSCs激活和EMT,提示LTBP2可能作为肝纤维化的潜在治疗靶点。