Induction of chemokines and cytokines before neutrophils and macrophage recruitment in different regions of rat liver after TAA administration
Induction of chemokines and cytokines before neutrophils and macrophage recruitment in different regions of rat liver after TAA administration
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DOI：
10.1038/labinvest.2013.134
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发表时间：
2014-02-01
影响因子：
5
通讯作者：
Malik, Ihtzaz A.
中科院分区：
文献类型：
--
作者：
Amanzada, Ahmad;Moriconi, Federico;Malik, Ihtzaz A.
单剂量硫代乙酰胺（TAA）给药诱导炎症和急性肝损伤。肝脏中炎症细胞募集的机制仍不清楚。本研究的目的是研究急性肝损伤过程中不同肝脏区域炎症细胞的序列和募集与CXC-和CC-趋化因子和细胞因子表达的关系。单剂量TAA腹腔内给予大鼠，然后在不同时间点处死动物。取血清和肝组织并立即冷冻。组织用于免疫染色低温切片、RNA和蛋白质提取。RT-PCR和蛋白质印迹分别进行RNA和蛋白质分析。与对照组相比，在TAA给药后，测量到CXCL 8/IL-8水平的早期增加（3小时），随后MCP 1/CCL 2血清水平（24小时）显著释放。类似地，在3 h发现肝趋化因子CXCL 1/KC和CXCL 8/IL-8的特异性RNA的早期增加，随后在24-48 h上调CXCL 5/LIX（6 h）、CXCL 2/MIP-2（12 h）和MCP 1/CCL 2基因表达。此外，观察到促炎细胞因子IFN-γ和IL-1 β的诱导，随后是IL-6和TNF-α，在12 h时达到最大值。在所有细胞因子和趋化因子中，TNF-α和MCP 1/CCL 2的基因表达增加幅度分别最高。通过免疫组织化学方法，观察到早期（12-24 h）仅中性粒细胞（NG）附着于门静脉血管壁及其周围的数量增加，随后沿着窦状隙单核吞噬细胞数量增加（24-48 h）。用TNF-α处理人单核细胞系U-937可增加CXCL 1/KC、CXCL 8/IL-8和MCP 1/CCL 2的基因表达。相反，在培养基中加入英夫利昔单抗（IFX）可显著抑制这种上调。总之，单剂量TAA给药诱导了一系列事件，其中炎性趋化因子和细胞因子的基因表达明确上调，NG在门脉区域和巨噬细胞沿着窦状隙在整个肝脏内短暂蓄积。门静脉周围炎症似乎先于肝细胞损伤。
Single-dose thioacetamide (TAA) administration induces inflammation and acute liver damage. The mechanism of inflammatory cell recruitment in the liver is still unclear. The aim of this study was to examine the sequence and recruitment of inflammatory cells in different liver regions in relation to CXC- and CC-chemokine and cytokine expression during acute liver injury. Single-dose TAA was administered to rats intraperitoneally, and animals were killed at different time points thereafter. Serum and liver tissue were taken and frozen immediately. Tissue was used for immunostaining cryostat sections, RNA, and protein extraction. RT-PCR and western blotting were performed for RNA and protein analysis, respectively. An early increase (3 h) in CXCL8/IL-8 levels was measured followed by a marked release in MCP1/CCL2 (24h) serum levels after TAA administration compared with controls. Similarly, an early increase in specific RNA of hepatic chemokines CXCL1/KC and CXCL8/IL-8 was found at 3 h, followed by an upregulation of CXCL5/LIX (6 h), CXCL2/MIP-2 (12 h), and MCP1/CCL2 gene expression at 24-48 h. Further, an induction of pro-inflammatory cytokines IFN-gamma and IL-1 beta followed by IL-6 and TNF-alpha was observed with a maximum at 12 h. The magnitude of increase in gene expression of TNF-alpha and MCP1/CCL2 was the highest among all cytokines and chemokines, respectively. By means of immunohistochemistry, an early (12-24 h) increase in the number of only neutrophil granulocytes (NGs) attached to and around portal vessel walls was observed, followed by increased numbers of mononuclear phagocytes (24-48 h) along the sinusoids. Treatment of the human monocytic cell line U-937 with TNF-alpha increased the gene expression of CXCL1/KC, CXCL8/IL-8, and MCP1/CCL2. Conversely, adding of infliximab (IFX) to the culture medium inhibited this upregulation significantly. In conclusion, single-dose TAA administration induces a sequence of events with a defined upregulation of gene expression of inflammatory chemokines and cytokines and a transient accumulation of NGs within the portal area and macrophages along the sinusoids throughout the liver. Periportal inflammation seems to precede hepatocellular damage.