Mátis G, Kulcsár A, Petrilla J, Talapka P, Neogrády Z
Department of Physiology and Biochemistry, Faculty of Veterinary Science, Szent István University, Budapest, Hungary.
J Anim Physiol Anim Nutr (Berl). 2017 Apr;101(2):201-207. doi: 10.1111/jpn.12547. Epub 2016 Jul 23.
As Kupffer cells are highly involved in the regulation of hepatic inflammatory response, the main goal of this study was to improve and to characterize a hepatocyte-Kupffer cell co-culture of pig origin for modelling endotoxin-induced hepatic inflammation and for testing the efficacy of potential anti-inflammatory substances. This monolayer co-culture was prepared from primary isolated swine hepatocytes and Kupffer cells in the ratio of 6:1 and 2:1, mimicking different states of liver inflammation. The prepared cell cultures were characterized by immunohistochemical CD-68 detection. Lipopolysaccharide (LPS) challenge of both co-cultures resulted in elevated interleukin-8 (IL-8) and that of 6:1 co-cultures in increased IL-6 production with a higher extent than on hepatocyte monocultures, justifying the key role of Kupffer cells in pro-inflammatory cytokine production. LPS-induced IL-8 production was successfully attenuated by concomitant application of both sodium butyrate and terpinen-4-ol on hepatocyte monocultures, but not on co-cultures, demonstrating the importance of the presence of Kupffer cells in cell cultures as inflammatory models. Based on these initial data, the applied porcine primary hepatocyte-Kupffer cell co-culture is suggested to be a proper tool for in vitro investigations on liver physiology and hepatic inflammation in pigs and can be used as a useful model mimicking in vivo conditions in veterinary research.
由于库普弗细胞高度参与肝脏炎症反应的调节，本研究的主要目标是改进并表征一种猪源肝细胞-库普弗细胞共培养体系，用于模拟内毒素诱导的肝脏炎症以及测试潜在抗炎物质的疗效。这种单层共培养体系由原代分离的猪肝细胞和库普弗细胞按6:1和2:1的比例制备而成，模拟不同的肝脏炎症状态。制备的细胞培养物通过免疫组织化学检测CD-68进行表征。两种共培养体系经脂多糖（LPS）刺激后，白细胞介素-8（IL-8）水平升高，6:1共培养体系中IL-6的产生增加，且增加程度高于肝细胞单培养体系，这证明了库普弗细胞在促炎细胞因子产生中的关键作用。在肝细胞单培养体系中，同时应用丁酸钠和萜品醇-4可成功减弱LPS诱导的IL-8产生，但在共培养体系中则不然，这表明在作为炎症模型的细胞培养物中，库普弗细胞的存在具有重要意义。基于这些初步数据，所应用的猪原代肝细胞-库普弗细胞共培养体系被认为是研究猪肝脏生理学和肝脏炎症的合适体外工具，可作为兽医研究中模拟体内条件的有用模型。
