Department of Medicine III, University Hospital Aachen, Pauwelsstrasse 30, D-52074 Aachen, Germany.
Gut. 2012 Mar;61(3):416-26. doi: 10.1136/gutjnl-2011-300304. Epub 2011 Aug 3.
Monocyte chemoattractant protein-1 (MCP-1, CCL2), the primary ligand for chemokine receptor C-C chemokine receptor 2 (CCR2), is increased in livers of patients with non-alcoholic steatohepatitis (NASH) and murine models of steatohepatitis and fibrosis. It was recently shown that monocyte/macrophage infiltration into the liver upon injury is critically regulated by the CCL2/CCR2 axis and is functionally important for perpetuating hepatic inflammation and fibrogenesis. The structured L-enantiomeric RNA oligonucleotide mNOX-E36 (a so-called Spiegelmer) potently binds and inhibits murine MCP-1. Pharmacological inhibition of MCP-1 with mNOX-E36 was investigated in two murine models of chronic liver diseases.
Pharmacological inhibition of MCP-1 by thrice-weekly mNOX-E36 subcutaneously was tested in murine models of acute or chronic carbon tetrachloride (CCl(4))- and methionine-choline-deficient (MCD) diet-induced chronic hepatic injury in vivo.
Antagonising MCP-1 by mNOX-E36 efficiently inhibited murine monocyte chemotaxis in vitro as well as migration of Gr1(+) (Ly6C(+)) blood monocytes into the liver upon acute toxic injury in vivo. In murine models of CCl(4)- and MCD diet-induced hepatic injury, the infiltration of macrophages into the liver was significantly decreased in anti-MCP-1-treated mice as found by fluorescence-activated cell sorting (FACS) analysis and immunohistochemistry. In line with lower levels of intrahepatic macrophages, proinflammatory cytokines (tumour necrosis factor α, interferon γ and interleukin 6) were significantly reduced in liver tissue. Overall fibrosis progression over 6 (CCl(4)) or 8 weeks (MCD diet) was not significantly altered by anti-MCP-1 treatment. However, upon MCD diet challenge a lower level of fatty liver degeneration (histology score, Oil red O staining, hepatic triglyceride content, lipogenesis genes) was detected in mNOX-E36-treated animals. mNOX-E36 also ameliorated hepatic steatosis upon therapeutic administration.
These results demonstrate the successful pharmacological inhibition of hepatic monocyte/macrophage infiltration by blocking MCP-1 during chronic liver damage in two in vivo models. The associated ameliorated steatosis development suggests that inhibition of MCP-1 is an interesting novel approach for pharmacological treatment in liver inflammation and steatohepatitis.
单核细胞趋化蛋白-1（MCP-1，CCL2）是趋化因子受体 C-C 趋化因子受体 2（CCR2）的主要配体，在非酒精性脂肪性肝炎（NASH）患者和脂肪性肝炎纤维化的小鼠模型的肝脏中增加。最近表明，损伤后单核细胞/巨噬细胞浸润肝脏受到 CCL2/CCR2 轴的严格调节，对于持续的肝炎症和纤维化具有功能重要性。结构 L-对映体 RNA 寡核苷酸 mNOX-E36（所谓的 Spiegelmer）强烈结合并抑制小鼠 MCP-1。用 mNOX-E36 对 MCP-1 进行药理学抑制在两种慢性肝病的小鼠模型中进行了研究。
用 mNOX-E36 每周三次皮下注射来抑制 MCP-1，在体内急性或慢性四氯化碳（CCl4）和蛋氨酸胆碱缺乏（MCD）饮食诱导的慢性肝损伤的小鼠模型中进行了研究。
用 mNOX-E36 拮抗 MCP-1 可有效抑制体外小鼠单核细胞趋化性以及体内急性毒性损伤时 Gr1（+）（Ly6C（+））血液单核细胞向肝脏的迁移。在 CCl4 和 MCD 饮食诱导的肝损伤的小鼠模型中，通过荧光激活细胞分选（FACS）分析和免疫组织化学发现，抗 MCP-1 治疗的小鼠肝脏内巨噬细胞的浸润明显减少。与肝内巨噬细胞水平降低相一致，肝组织中的促炎细胞因子（肿瘤坏死因子-α、干扰素-γ和白细胞介素 6）显著减少。在 6 周（CCl4）或 8 周（MCD 饮食）的总体纤维化进展中，抗 MCP-1 治疗并没有明显改变。然而，在用 MCD 饮食挑战时，在用 mNOX-E36 治疗的动物中检测到较低水平的脂肪肝变性（组织学评分、油红 O 染色、肝甘油三酯含量、脂肪生成基因）。mNOX-E36 还改善了治疗给药时的肝脂肪变性。
这些结果表明，在两种体内模型中，通过阻断 MCP-1 在慢性肝损伤期间成功地抑制了肝单核细胞/巨噬细胞的浸润。相关的脂肪变性发展的改善表明，抑制 MCP-1 是治疗肝炎症和脂肪性肝炎的一种有前途的新方法。
