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目的 探究褪黑素是否通过调控Wnt/β-连环蛋白(Wnt/β-catenin)信号通路发挥抗肝纤维化作用。方法 在四氯化碳(CCL4)诱导的小鼠肝纤维化模型中，利用HE染色观察肝脏组织形态，天狼星红染色评估胶原沉积水平，免疫组化检测肝组织α-平滑肌肌动蛋白(α-SMA)、Wnt家族成员1(Wnt1)、β-连环蛋白(β-catenin)蛋白水平，免疫蛋白印迹检测α-SMA、Ⅰ型胶原(COL1)、Wnt1、β-catenin、骨髓细胞瘤病毒癌基因同源物(c-MYC)及细胞周期蛋白D1(CyclinD1)蛋白表达水平，qRT-PCR法检测Wnt1和β-cateninmRNA表达水平。在体外激活的肝星状细胞中，加入β-catenin激活剂后采用CCK-8检测肝星状细胞增殖水平，免疫蛋白印迹检测α-SMA、COL1、β-catenin、c-MYC和CyclinD1蛋白表达水平，qRT-PCR法检测COL1、α-SMA、c-MYC及CyclinD1mRNA表达水平。结果 体内实验中，CCL4处理的小鼠肝纤维化相关指标α-SMA、COL1水平及Wnt/β-catenin信号通路相关蛋白表达升高(P<0.05)，褪黑素干预后肝纤维化水平减轻明显，上述指标均明显下调(P<0.05)。在体外培养的肝星状细胞中，激活后的肝星状细胞增殖明显，同时Wnt/β-catenin信号通路相关蛋白表达明显升高(P<0.05)，褪黑素处理后肝星状细胞增殖活性及Wnt/β-catenin信号通路相关蛋白的表达均明显降低(P<0.05)。而在β-catenin激活剂处理后的肝星状细胞中，褪黑素对肝星状细胞增殖、活化以及Wnt/β-catenin信号通路抑制作用均部分逆转(P<0.05)。结论 褪黑素通过调控Wnt/β-catenin通路抑制肝星状细胞激活，发挥抗肝纤维化作用，为其治疗肝纤维化提供理论依据。
Abstract：Objective To investigate whether melatonin exerts anti-hepatic fibrosis effects by regulating the Wnt/β-catenin signaling pathway. Methods In a carbon tetrachloride (CCl4) -induced mouse hepatic fibrosis model， HE staining was used to observe the morphological changes of liver tissue. Sirius Red staining was applied to evaluate the level of collagen deposition. Immunohistochemistry was performed to detect the protein levels of α-smooth muscle actin (α-SMA)， Wnt family member 1 (Wnt1) and β-catenin in liver tissues. Western blot was adopted to detect the protein expression levels of α-SMA， type Ⅰ collagen (COL1)， Wnt1， β-catenin， myelocytomatosis viral oncogene homolog (c-MYC) and cyclin D1， and qRT-PCR was used to detect the mRNA expression levels of Wnt1 and β-catenin. In activated hepatic stellate cells in vitro， after treatment with a β-catenin activator， CCK-8 assay was used to detect the proliferation of hepatic stellate cells. Western blot was applied to measure the protein expression levels of α-SMA， COL1， β-catenin， c-MYC and cyclin D1， and qRT-PCR was used to detect the mRNA expression levels of COL1， α-SMA， c-MYC and cyclin D1. Results The levels of hepatic fibrosis-related indicators α-SMA and COL1， as well as the expression of proteins related to the Wnt/β-catenin signaling pathway， increased in CCl4-treated mice (P<0.05). Melatonin intervention significantly alleviated liver fibrosis and downregulated the expression of these indicators (P<0.05). In vitro， activated hepatic stellate cells (HSCs) exhibited enhanced proliferative activity and increased expression of Wnt/β-catenin signaling pathway-related proteins (P<0.05). Melatonin treatment significantly reduced HSCs proliferation and suppressed the expression of Wnt/β-catenin signaling pathway-related proteins (P<0.05). Furthermore， treatment with a β-catenin activator partially reversed the inhibitory effects of melatonin on HSCs proliferation， activation， and Wnt/β-catenin signaling pathway activity (P<0.05). Conclusion Melatonin suppresses hepatic stellate cell activation via regulating the Wnt/β-catenin pathway， thereby exerting anti-fibrotic effects， which provides a theoretical basis for its application in hepatic fibrosis treatment.
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基本信息:
中图分类号:R575.2
引用信息:
[1]揭磊,黄启鸣,刘丝丝,等.基于Wnt/β-catenin信号通路探究褪黑素抗肝纤维化作用[J].安徽医科大学学报().
基金信息:
国家自然科学基金项目（编号：81700521）; 安徽省重点研究与开发计划项目（编号：202004j07020039）
2026-09-05
2026-09-05
2026-09-05