Knockout of the Tachykinin Receptor 1 in the Mdr2-/- (Abcb4-/-) Mouse Model of Primary Sclerosing Cholangitis Reduces Biliary Damage and Liver Fibrosis
Knockout of the Tachykinin Receptor 1 in the Mdr2-/- (Abcb4-/-) Mouse Model of Primary Sclerosing Cholangitis Reduces Biliary Damage and Liver Fibrosis
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DOI：
10.1016/j.ajpath.2020.07.007
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发表时间：
2020-11-01
影响因子：
6
通讯作者：
Alpini, Gianfranco
中科院分区：
文献类型：
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作者：
Ceci, Ludovica;Francis, Heather;Alpini, Gianfranco
P 物质 (SP)/神经激肽 1 受体 (NK1R) 轴的激活通过增强转化生长因子-β 1 (TGF-β 1) 胆汁分泌，触发胆管结扎小鼠和 Mdr2(-/-)（别名 Abcb4(-/-)）小鼠的胆道损伤/衰老和肝纤维化。最近的证据表明 miR-31 (MIR31) 在 TGF-β1 诱导的肝纤维化中发挥作用。我们的目的是确定 SP/NK1R/TGF-β 1/miR-31 轴在胆汁淤积期间调节胆道增殖和肝纤维化中的作用。因此，我们生成了一个双敲除 Mdr2(-/-) 和 NK1R(-/)（别名 Tacr1(-/-)）的新模型，以进一步探讨 SP/NK1R 轴在慢性胆汁淤积期间的作用。在以下 12 周龄雄性小鼠中进行了体内研究：(i) NK1R(-/-)； (ii) Mdr2(-/-)； (iii) NK1R(-/-)/Mdr2(-/-) (Tacr1(-/-)/Abcb4(-/-)) 及其相应的野生型对照。收集肝组织和胆管细胞，通过肝切片免疫组织化学和实时PCR评估肝损伤、胆汁肿块/衰老的变化、炎症和肝纤维化。通过实时 PCR 测量分离胆管细胞中的 miR-31 表达。与Mdr2(-/-)小鼠相比，NK1R(-/-)/Mdr2(-/-)小鼠的胆管反应、肝纤维化、胆道衰老和胆道炎症减少。在Mdr2(-/-)小鼠中观察到miR-31表达升高，在NK1R(-/-)/Mdr2(-/-)小鼠中观察到miR-31表达降低。靶向 SP/NK1R 和/或 miR-31 可能是治疗人类胆管病（包括原发性硬化性胆管炎）的潜在方法。
Activation of the substance P (SP)/neurokinin 1 receptor (NK1R) axis triggers biliary damage/senescence and liver fibrosis in bile duct ligated and Mdr2(-/-) (alias Abcb4(-/-)) mice through enhanced transforming growth factor-beta 1 (TGF-beta 1) biliary secretion. Recent evidence indicates a role for miR-31 (MIR31) in TGF-beta 1-induced liver fibrosis. We aimed to define the role of the SP/NK1R/TGF-beta 1/miR-31 axis in regulating biliary proliferation and liver fibrosis during cholestasis. Thus, we generated a novel model with double knockout of Mdr2(-/-) and NK1R(-/) (alias Tacr1(-/-)) to further address the role of the SP/NK1R axis during chronic cholestasis. In vivo studies were performed in the following 12-week-old male mice: (i) NK1R(-/-); (ii) Mdr2(-/-); and (iii) NK1R(-/-)/Mdr2(-/-) (Tacr1(-/-)/Abcb4(-/-)) and their corresponding wild-type controls. Liver tissues and cholangiocytes were collected, and liver damage, changes in biliary mass/senescence, and inflammation as well as liver fibrosis were evaluated by both immunohistochemistry in liver sections and real-time PCR. miR-31 expression was measured by real-time PCR in isolated cholangiocytes. Decreased ductular reaction, liver fibrosis, biliary senescence, and biliary inflammation were observed in NK1R(-/-)/Mdr2(-/-) mice compared with Mdr2(-/-) mice. Elevated expression of miR-31 was observed in Mdr2(-/-) mice, which was reduced in NK1R(-/-)/Mdr2(-/-) mice. Targeting the SP/NK1R and/or miR-31 may be a potential approach in treating human cholangiopathies, including primary sclerosing cholangitis.