Department of Gastroenterology, The Second Affiliated Hospital of Nanchang University, Nanchang 330006, China.
Medical College of Nanchang University, Nanchang 330006, China.
Gastroenterol Hepatol. 2024 Mar;47(3):219-229. doi: 10.1016/j.gastrohep.2023.05.005. Epub 2023 May 18.
Hepatic fibrosis (HF) is a common result of the repair process of various chronic liver diseases. Hepatic stellate cells (HSCs) activation is the central link in the occurrence of HF.
ELISA and histological analysis were performed to detect the pathological changes of liver tissues. In vitro, HSCs were treated with TGF-β1 as HF cell model. Combination of GATA-binding protein 3 (GATA3) and miR-370 gene promoter was ensured by ChIP and luciferase reporter assay. Autophagy was monitored by observing the GFP-LC3 puncta formation. The interaction between miR-370 and high mobility group box 1 protein (HMGB1) was verified by luciferase reporter assay.
CCl-induced HF mice exhibited an increase of ALT and AST, and severe damage and fibrosis of liver tissues. GATA3 and HMGB1 were up-regulated, and miR-370 was down-regulated in CCl-induced HF mice and activated HSCs. GATA3 enhanced expression of the autophagy-related proteins and activation markers in the activated HSCs. Inhibition of autophagy partly reversed GATA3-induced activation of HSCs and the promotion of GATA3 to hepatic fibrosis. Moreover, GATA3 suppressed miR-370 expression via binding with its promotor, and enhanced HMGB1 expression in HSCs. Increasing of miR-370 inhibited HMGB1 expression by directly targeting its mRNA 3'-UTR. The promotion of GATA3 to TGF-β1-induced HSCs autophagy and activation was abrogated by miR-370 up-regulation or HMGB1 knockdown.
This work demonstrates that GATA3 promotes autophagy and activation of HSCs by regulating miR-370/HMGB1 signaling pathway, which contributes to accelerate HF. Thus, this work suggests that GATA3 may be a potential target for prevention and treatment of HF.
肝纤维化（HF）是各种慢性肝病修复过程中的常见结果。肝星状细胞（HSCs）的激活是 HF 发生的中心环节。
采用 ELISA 和组织学分析检测肝组织的病理变化。体外，用 TGF-β1 处理 HSCs 作为 HF 细胞模型。采用 ChIP 和荧光素酶报告基因检测确保 GATA 结合蛋白 3（GATA3）和 miR-370 基因启动子的结合。通过观察 GFP-LC3 斑点的形成来监测自噬。通过荧光素酶报告基因检测验证 miR-370 与高迁移率族蛋白 B1（HMGB1）之间的相互作用。
CCl 诱导的 HF 小鼠表现出 ALT 和 AST 升高，肝组织严重损伤和纤维化。在 CCl 诱导的 HF 小鼠和激活的 HSCs 中，GATA3 和 HMGB1 上调，而 miR-370 下调。GATA3 增强了激活的 HSCs 中自噬相关蛋白和激活标志物的表达。自噬抑制部分逆转了 GATA3 诱导的 HSCs 激活和对肝纤维化的促进作用。此外，GATA3 通过与启动子结合抑制 miR-370 的表达，并增强 HSCs 中的 HMGB1 表达。miR-370 通过直接靶向其 mRNA 3'-UTR 抑制 HMGB1 表达。miR-370 上调或 HMGB1 敲低可消除 GATA3 对 TGF-β1 诱导的 HSCs 自噬和激活的促进作用。
本研究表明，GATA3 通过调节 miR-370/HMGB1 信号通路促进 HSCs 的自噬和激活，有助于加速 HF。因此，本研究表明 GATA3 可能是预防和治疗 HF 的潜在靶点。
