Mito Masaki, Tsuchiya Atsunori, Ishii Soichi, Tonouchi Takafumi, Furuyama Kaito, Jinbo Ryo, Takeda Nobutaka, Abe Hiroyuki, Tamai Katsuto, Terai Shuji
Division of Gastroenterology and Hepatology, Graduate School of Medical and Dental Sciences, Niigata University, 1-757, Asahimachi-dori, Chuo-ku, Niigata, 951-8510, Japan.
Department of Stem Cell Therapy Science, Graduate School of Medicine, Osaka University, 2-2, Yamadaoka, Suita, Osaka, 565-0871, Japan.
Biochem Biophys Rep. 2025 May 25;42:102061. doi: 10.1016/j.bbrep.2025.102061. eCollection 2025 Jun.
Liver cirrhosis is a serious disease characterized by liver dysfunction and severe fibrosis; however, no breakthrough drugs have effectively improved fibrosis, making it an unmet medical need. We have previously reported that the HMGB1 peptide, synthesized from box A of the HMGB1 protein, ameliorates liver fibrosis and is a promising candidate for fibrosis-improving drugs against liver cirrhosis. In this study, we used spatial analysis to observe treatment-induced changes over time.
Liver cirrhosis was induced in C57BL/6J mice using carbon tetrachloride (CCl4) injections, followed by HMGB1 peptide treatment. To assess the temporal effects of HMGB1 on the liver in a CCl4-induced cirrhosis mouse model, we used GeoMx spatial analysis. We focused on αSMA-positive active hepatic stellate cells (HSCs), F4/80-positive macrophages, and CK8/18-positive hepatocytes to determine how each cell type was affected over time. Statistical analyses were conducted using GraphPad Prism9, with significance set at p < 0.05.
In cirrhotic mice, we first observed a decrease in the number of activated HSCs over time, two weeks after treatment initiation. Macrophage-associated genes ceased to induce fibrosis-related pathways early in the treatment. This suggests that the effect of macrophages on fibrosis was weakened by the treatment. We also confirmed that lipid metabolism of hepatocytes may be improved during treatment. Furthermore, and expression were induced in the peptide-treated group, indicating possible cell migration to the liver.
Over time, macrophages followed by HSCs, showed the most notable changes with treatment, resulting in improved fibrosis. The HMGB1 peptide drug also affected lipid metabolism in hepatocytes, suggesting a positive therapeutic effect on steatohepatitis. Elevated factors that promote cell migration may have also enhanced the healing effect.
肝硬化是一种以肝功能障碍和严重纤维化为特征的严重疾病；然而，尚无突破性药物能有效改善纤维化，这使其成为未满足的医疗需求。我们之前报道过，由高迁移率族蛋白B1（HMGB1）蛋白的A盒合成的HMGB1肽可改善肝纤维化，是治疗肝硬化的抗纤维化药物的一个有前景的候选药物。在本研究中，我们使用空间分析来观察治疗随时间引起的变化。
通过注射四氯化碳（CCl4）在C57BL/6J小鼠中诱导肝硬化，随后进行HMGB1肽治疗。为了评估HMGB1对CCl4诱导的肝硬化小鼠模型肝脏的时间效应，我们使用了GeoMx空间分析。我们重点关注α平滑肌肌动蛋白（αSMA）阳性的活化肝星状细胞（HSCs）、F4/80阳性的巨噬细胞和细胞角蛋白8/18（CK8/18）阳性的肝细胞，以确定每种细胞类型随时间如何受到影响。使用GraphPad Prism9进行统计分析，显著性设定为p < 0.05。
在肝硬化小鼠中，我们首先观察到在开始治疗两周后，活化HSCs的数量随时间减少。巨噬细胞相关基因在治疗早期停止诱导纤维化相关途径。这表明治疗削弱了巨噬细胞对纤维化的作用。我们还证实，在治疗期间肝细胞的脂质代谢可能得到改善。此外，在肽治疗组中诱导了[此处原文缺失具体基因名称]和[此处原文缺失具体基因名称]的表达，表明可能有细胞迁移至肝脏。
随着时间推移，巨噬细胞随后是HSCs，显示出治疗引起的最显著变化，从而改善了纤维化。HMGB1肽药物还影响了肝细胞的脂质代谢，表明对脂肪性肝炎有积极的治疗作用。促进细胞迁移的升高因子可能也增强了愈合效果。
