Carvedilol Improves Inflammatory Response, Oxidative Stress and Fibrosis in the Alcohol-Induced Liver Injury in Rats by Regulating Kuppfer Cells and Hepatic Stellate Cells.
Carvedilol Improves Inflammatory Response, Oxidative Stress and Fibrosis in the Alcohol-Induced Liver Injury in Rats by Regulating Kuppfer Cells and Hepatic Stellate Cells.
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DOI：
10.1371/journal.pone.0148868
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
de Araújo AA
中科院分区：
文献类型：
--
作者：
Araújo Júnior RF;Garcia VB;Leitão RF;Brito GA;Miguel Ede C;Guedes PM;de Araújo AA
探讨卡维地洛（carvedilol，CARV）对酒精性肝损伤大鼠的抗炎、抗氧化及抗纤维化作用。连续28天灌胃乙醇（7 g/kg）诱导肝损伤。80只Wistar大鼠在暴露于酒精前1 h用1、3或5 mg/kg的口服CARV或生理盐水预处理。检测肝组织匀浆中白细胞介素（IL）-1β、IL-10、肿瘤坏死因子（TNF）-α的含量，以及髓过氧化物酶（MPO）活性、丙二醛（MDA）和谷胱甘肽（GSH）的含量。测定血清谷草转氨酶（AST）活性和肝脏甘油三酯（TG）水平。对环氧合酶2（考克斯-2）、核因子κ B/配体受体激活剂（RANK/RANKL）、细胞因子信号传导抑制因子（SOCS 1）、枯否细胞标志物IBA-1（离子钙结合衔接分子1）、细胞间粘附分子1（ICAM-1）、超氧化物歧化酶（SOD-1）和谷胱甘肽过氧化物酶（GPx-1）表达进行免疫组织化学分析。共聚焦显微镜分析IL-1β和NF-κB表达，实时定量PCR分析TNFα、PCI、PC III和NF-κB。在酒精暴露方案期间，CARV治疗（5 mg/kg）与脂肪变性、肝索变性、纤维化和坏死减少以及AST水平降低相关。（p < 0.01），ALT（p < 0.01），TG（p < 0.001），MPO（p < 0.001），MDA（p < 0.05），和促炎细胞因子（IL-1β和TNF-α，均p < 0.05），抗炎细胞因子IL-10（p < 0.001）和GSH（p < 0.05）水平增加。CARV 5 mg/kg治疗还降低了考克斯-2、RANK、RANKL、IBA-1和ICAM-1的表达水平（均p < 0.05），同时增加了SOCS 1、SOD-1和GPx-1的表达（均p < 0.05），并降低了IL-1β和NF-κB的表达（均p < 0.05）。实时定量PCR分析显示，相对于仅酒精组，酒精-CARV 5 mg/kg组中TNF-α、I型前胶原（PCI）、III型前胶原（PCIII）和NF-κB的mRNA产生减少。CARV可通过抑制炎性细胞因子下调枯否细胞和肝星状细胞（HSC）的信号传导，减少大鼠模型中乙醇诱导的肝损伤中的应激氧化、炎症反应和纤维化。
To evaluate the anti-inflammatory, anti-oxidant and antifibrotic effects of carvedilol (CARV) in rats with ethanol-induced liver injury. Liver injury was induced by gavage administration of alcohol (7 g/kg) for 28 consecutive days. Eighty Wistar rats were pretreated with oral CARV at 1, 3, or 5 mg/kg or with saline 1 h before exposure to alcohol. Liver homogenates were assayed for interleukin (IL)-1β, IL-10, and tumor necrosis factor (TNF)-α level as well as for myeloperoxidase (MPO) activity and malonyldialdehyde (MDA) and glutathione (GSH) levels. Serum aspartate aminotransferase (AST) activity and liver triglyceride (TG) levels were also assayed. Immunohistochemical analyses of cyclooxygenase 2 (COX-2), receptor activator of nuclear factor kappa-B/ligand (RANK/RANKL), suppressor of cytokine signalling (SOCS1), the Kupffer cell marker IBA-1 (ionized calcium-binding adaptor molecule 1), intercellular adhesion molecule 1 (ICAM-1), superoxide dismutase (SOD-1), and glutathione peroxidase (GPx-1) expression were performed. Confocal microscopy analysis of IL-1β and NF-κB expression and real-time quantitative PCR analysis for TNFα, PCI, PCIII, and NF-κB were performed. CARV treatment (5 mg/kg) during the alcohol exposure protocol was associated with reduced steatosis, hepatic cord degeneration, fibrosis and necrosis, as well as reduced levels of AST (p < 0.01), ALT (p < 0.01), TG (p < 0.001), MPO (p < 0.001), MDA (p < 0.05), and proinflammatory cytokines (IL-1β and TNF-α, both p < 0.05), and increased levels of the anti-inflammatory cytokine IL-10 (p < 0.001) and GSH (p < 0.05), compared to the alcohol-only group. Treatment with CARV 5 mg/kg also reduced expression levels of COX-2, RANK, RANKL, IBA-1, and ICAM-1 (all p < 0.05), while increasing expression of SOCS1, SOD-1, and GPx-1 (all p < 0.05) and decreasing expression of IL-1β and NF-κB (both, p < 0.05). Real-time quantitative PCR analysis showed that mRNA production of TNF-α, procollagen type I (PCI), procollagen type III (PCIII), and NF-κB were decreased in the alcohol-CARV 5 mg/kg group relative to the alcohol-only group. CARV can reduce the stress oxidative, inflammatory response and fibrosis in ethanol-induced liver injury in a rat model by downregulating signalling of Kuppfer cells and hepatic stellate cells (HSCs) through suppression of inflammatory cytokines.