The inhibition of YAP Signaling Prevents Chronic Biliary Fibrosis in the Abcb4(-/-) Model by Modulation of Hepatic Stellate Cell and Bile Duct Epithelium Cell Pathophysiology.
The inhibition of YAP Signaling Prevents Chronic Biliary Fibrosis in the Abcb4(-/-) Model by Modulation of Hepatic Stellate Cell and Bile Duct Epithelium Cell Pathophysiology.
复制标题
DOI：
10.14336/ad.2023.0602
复制
发表时间：
2024-02-01
影响因子：
7.4
通讯作者：
中科院分区：
文献类型：
--
作者：
原发性硬化性胆管炎（PSC）是一种慢性肝病，其特征是预后不良，缺乏病因治疗选择。是相关蛋白（雅普）的功能作为一个重要的调解人的纤维化，然而，其治疗潜力的慢性胆道疾病，如PSC仍然没有建立。本研究的目的是通过检测肝星状细胞（HSC）和胆管上皮细胞（BEC）的病理生理学来阐明雅普抑制在胆管纤维化中的可能意义。分析来自PSC患者的人肝组织样品以评估相对于非纤维化对照样品的雅普/结缔组织生长因子（CTGF）的表达。在原代人HSC（phHSC）、LX-2、H69和TFK-1细胞系中通过siRNA或使用维替泊芬（VP）和二甲双胍（MF）的药理学抑制研究了HSC和BEC中雅普/CTGF的病理生理学相关性。采用Abcb 4-/-小鼠模型评价药理学雅普抑制的保护作用。采用悬滴法和三维基质胶培养技术研究不同物理条件下phHSC雅普的表达和活化状态。在PSC患者中观察到雅普/CTGF上调。沉默雅普/CTGF导致phHSC活化的抑制和LX-2细胞收缩性的降低，以及H69细胞中的上皮-间充质转化（EMT）和TFK-1细胞的增殖的抑制。药理学抑制雅普减轻体内慢性肝纤维化，减少小管反应和EMT。phHSC中的雅普表达通过改变细胞外刚度而被有效地调节，突出了雅普作为机械换能器的作用。总之，雅普调节BEC中HSC和EMT的活化，从而作为慢性胆汁淤积中纤维化的检查点。VP和MF均显示出作为雅普抑制剂的有效性，能够抑制胆汁纤维化。这些结果表明，VP和MF作为治疗PSC的潜在治疗选择值得进一步研究。
Primary sclerosing cholangitis (PSC) represents a chronic liver disease characterized by poor prognosis and lacking causal treatment options. Yes-associated protein (YAP) functions as a critical mediator of fibrogenesis; however, its therapeutic potential in chronic biliary diseases such as PSC remains unestablished. The objective of this study is to elucidate the possible significance of YAP inhibition in biliary fibrosis by examining the pathophysiology of hepatic stellate cells (HSC) and biliary epithelial cells (BEC). Human liver tissue samples from PSC patients were analyzed to assess the expression of YAP/connective tissue growth factor (CTGF) relative to non-fibrotic control samples. The pathophysiological relevance of YAP/CTGF in HSC and BEC was investigated in primary human HSC (phHSC), LX-2, H69, and TFK-1 cell lines through siRNA or pharmacological inhibition utilizing verteporfin (VP) and metformin (MF). The Abcb4-/- mouse model was employed to evaluate the protective effects of pharmacological YAP inhibition. Hanging droplet and 3D matrigel culture techniques were utilized to investigate YAP expression and activation status of phHSC under various physical conditions. YAP/CTGF upregulation was observed in PSC patients. Silencing YAP/CTGF led to inhibition of phHSC activation and reduced contractility of LX-2 cells, as well as suppression of epithelial-mesenchymal transition (EMT) in H69 cells and proliferation of TFK-1 cells. Pharmacological inhibition of YAP mitigated chronic liver fibrosis in vivo and diminished ductular reaction and EMT. YAP expression in phHSC was effectively modulated by altering extracellular stiffness, highlighting YAP's role as a mechanotransducer. In conclusion, YAP regulates the activation of HSC and EMT in BEC, thereby functioning as a checkpoint of fibrogenesis in chronic cholestasis. Both VP and MF demonstrate effectiveness as YAP inhibitors, capable of inhibiting biliary fibrosis. These findings suggest that VP and MF warrant further investigation as potential therapeutic options for the treatment of PSC.