HSC-specific inhibition of Rho-kinase reduces portal pressure in cirrhotic rats without major systemic effects
HSC-specific inhibition of Rho-kinase reduces portal pressure in cirrhotic rats without major systemic effects
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DOI：
10.1016/j.jhep.2012.07.033
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发表时间：
2012-12-01
影响因子：
25.7
通讯作者：
Trebicka, Jonel
中科院分区：
文献类型：
--
作者：
Klein, Sabine;Van Beuge, Marike Marjolijn;Trebicka, Jonel
背景和目标：Rho激酶激活介导细胞收缩，增加肝内阻力，从而增加肝硬化门静脉压力。系统性Rho激酶抑制降低肝硬化门静脉压，但也降低动脉压。因此，需要肝脏特异性Rho激酶抑制。Rho激酶抑制剂对活化的肝星状细胞的递送减少了纤维化。它也可能放松这些收缩细胞，从而降低肝内阻力。方法：肝硬化模型采用四氯化碳中毒和胆管结扎。注射Rho激酶抑制剂（Y26732）与靶向激活的肝星状细胞的载体（甘露糖-6-磷酸修饰的人血清白蛋白）偶联后3小时，通过彩色微球技术和直接压力测量分析血流动力学。通过免疫组化染色和Western blot检测Rho激酶抑制剂的作用部位和作用效果。Rho激酶抑制剂与未修饰的人血清白蛋白偶联的实验作为非靶向control.Results：在两种肝硬化模型中，载体偶联Rho激酶抑制剂降低了门静脉压力，降低了肝门静脉阻力。免疫组化结蛋白染色显示肝星状细胞中的载体。靶向治疗降低了Rho激酶（膜突蛋白）磷酸化底物的表达，并消除了纤维化隔膜中的肌球蛋白轻链磷酸化（胶原染色）。靶向Rho激酶抑制剂未显示出重大肝外效应。相比之下，非靶向Rho激酶抑制剂引起严重的全身性hypothesis.Conclusions：肝星状细胞活化是至关重要的参与肝硬化门静脉高压症。如前所述，以肝星状细胞中的Rhokinase为靶点不仅可以减少纤维化，而且可以急性降低门静脉压力，而不会产生主要的全身效应。(C)2012年欧洲肝脏研究协会。Elsevier B. V.出版，保留所有权利。
Background & Aims: Rho-kinase activation mediates cell contraction and increases intrahepatic resistance and consequently portal pressure in liver cirrhosis. Systemic Rho-kinase inhibition decreases portal pressure in cirrhosis, but also arterial pressure. Thus, liver-specific Rho-kinase inhibition is needed. The delivery of Rho-kinase inhibitor to activated hepatic stellate cells reduces fibrosis. It might also relax these contractile cells and therewith decrease intrahepatic resistance. We tested this hypothesis by performing acute experiments in cirrhotic rats.Methods: Cirrhosis models were CCI4-intoxication and bile duct ligation. Three hours after injection of the Rho-kinase inhibitor (Y26732) coupled with a carrier (mannose-6-phosphate modified human serum albumin), which targets activated hepatic stellate cells, hemodynamics were analyzed by the colored microsphere technique and direct pressure measurements. The delivery site and effect of Rho-kinase inhibitor were investigated by immunohistochemical stainings, as well as Western blot. Experiments with Rho-kinase inhibitor coupled with unmodified human serum albumin served as untargeted control.Results: In both models of cirrhosis, the carrier coupled Rhokinase inhibitor lowered the portal pressure and decreased the hepatic-portal resistance. Immunohistochemical desmin-staining showed the carrier in hepatic stellate cells. The targeted therapy decreased the expression of the phosphorylated substrate of Rho-kinase (moesin) and abolished myosin light chains phosphorylation in fibrotic septae (collagen-staining). The targeted Rho-kinase inhibitor showed no major extrahepatic effects. By contrast, the untargeted Rho-kinase inhibitor elicited severe systemic hypotension.Conclusions: Activated hepatic stellate cells are crucially involved in portal hypertension in cirrhosis. Targeting of Rhokinase in hepatic stellate cells not only decreased fibrosis, as previously shown, but also lowers portal pressure acutely without major systemic effects as demonstrated in this study. (C) 2012 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.