Cytokine‐induced neutrophil chemoattractant release from hepatocytes is modulated by Kupffer cells
Cytokine‐induced neutrophil chemoattractant release from hepatocytes is modulated by Kupffer cells
复制标题
细胞因子诱导的中性粒细胞趋化剂从肝细胞中释放受库普弗细胞调节
作者：
E. Mawet;Y. Shiratori;Yohko Hikiba;H. Takada;Hideo Yoshida;K. Okano;Y. Komatsu;M. Matsumura;Y. Niwa;Masao Omata
为了阐明中性粒细胞蓄积期间肝脏中细胞间通讯的作用，在体外Kupffer细胞条件培养基存在下研究了肝细胞释放细胞因子诱导的中性粒细胞趋化因子（CINC）（啮齿动物中的白细胞介素-8 [IL-8]相关蛋白）。通过用胶原酶灌注大鼠肝脏，然后在甲泛葡胺梯度上离心来制备枯否细胞，并在存在或不存在脂多糖（LPS）的情况下进行培养。24小时后收集条件培养基，并在存在或不存在库普弗细胞条件培养基的情况下培养大鼠肝细胞。通过蛋白质印迹分析和酶联免疫吸附测定（ELISA）测量培养上清液中的CINC的量，并通过聚合酶链反应评估其信使RNA（mRNA）的表达。LPS刺激的枯否细胞条件培养基增强了肝细胞中CINC mRNA的表达，并增加了肝细胞产生CINC的能力。当库普弗细胞条件培养基用热预处理（56 ℃，30分钟）时，未显示CINC的产生增强。在存在LPS刺激的库普弗细胞条件培养基的情况下，肝细胞产生的CINC被抗白细胞介素1β（IL-1β）抗体减少，但不被抗肿瘤坏死因子α（TNF-α）或LPS抗体减少。这些结果表明，肝细胞产生的CINC可受枯否细胞释放的IL-1β调节，导致肝损伤期间中性粒细胞蓄积，因为该蛋白质是中性粒细胞的强化学引诱物。
To clarify the role of intercellular communication in the liver during accumulation of neutrophils, the release of cytokine‐induced neutrophil chemoattractant (CINC) (interleukin‐8 [IL‐8] related protein in rodents) by hepatocytes was investigated in the presence of Kupffer cell‐conditioned medium in vitro. Kupffer cells were prepared by perfusion of rat liver with collagenase followed by centrifugation on a metrizamide gradient and were cultured in the presence or absence of lipopolysacharide (LPS). The conditioned medium was collected after 24 hours, and rat hepatocytes were cultured in the presence or absence of Kupffer cell‐conditioned medium. An amount of CINC in the culture supernatant was measured by western blotting analysis and enzyme‐linked immunosorbent assay (ELISA), and expression of its messenger RNA (mRNA) was assessed by the polymerase chain reaction. LPS‐stimulated Kupffer cell‐conditioned medium enhanced an expression of CINC mRNA in hepatocytes and increased the production of CINC by hepatocytes. Enhanced production of CINC was not shown when the Kupffer cell‐conditioned medium was pretreated with heat (56 degrees C, 30 minutes). The production of CINC by hepatocytes in the presence of the LPS‐stimulated Kupffer cell‐conditioned medium was reduced by an antibody against interleukin 1β (IL‐1β), but not by antibodies against tumor necrosis factor α (TNF‐α) or LPS. These results suggest that production of CINC by hepatocytes could be regulated by IL‐1β released from Kupffer cells, leading to neutrophil accumulation during liver injury, because this protein is a strong chemoattractant for neutrophils.