Zocco M A, Carloni E, Pescatori M, Saulnier N, Lupascu A, Nista E C, Novi M, Candelli M, Cimica V, Mihm S, Gasbarrini G, Ramadori G, Gasbarrini A
Department of Internal Medicine, Catholic University of Rome, Italy.
Dig Liver Dis. 2006 Aug;38(8):563-77. doi: 10.1016/j.dld.2006.04.015. Epub 2006 Jun 27.
Kupffer cells are intrasinusoidal space located macrophages with phagocytic capacity. Interferons are cytokines with antiviral, antiproliferative and immunomodulatory activities which may influence the activity of Kupffer cells. Aim of this study was to evaluate Kupffer cell gene expression after interferon-alpha or interferon-gamma stimulation in order to investigate a link between these cytokines and macrophage activation.
Rat Kupffer cells were cultured for 24 h and divided into three groups: unstimulated; stimulated with interferon-alpha and stimulated with interferon-gamma. After 8 h stimulation total RNA was extracted and processed according to Affymetrix protocols and hybridised on R34A microarray gene set. Data analyses was performed using Microarray Analysis Suite 5.0 software. Genes showing remarkably different expression in microarray analysis were confirmed by real-time PCR.
Nearly 4000 out of the 8800 genes represented in the array were expressed by Kupffer cells. Among these, interferon-alpha up-regulates 91 genes by over two-fold (antiviral, antigen processing and presentation, and tumour suppressor/proapoptotic genes) and down-regulates 72 genes by 50% or more. Interferon-gamma up-regulates 70 genes by over two-fold and down-regulates 78 genes by 50% or more. Most of the genes induced by interferon-alpha are also induced by interferon-gamma. Down-regulated genes include growth factors and genes involved in cell cycle/proliferation. Real-time PCR confirms the results of the array.
Interferons directly target rat Kupffer cells and are involved in the regulation of a wide variety of genes. Their expression profile shed light onto molecular mechanism of Kupffer cells activation in specific pathways such as antiviral and antitumour processes.
库普弗细胞是位于肝血窦内具有吞噬能力的巨噬细胞。干扰素是一类具有抗病毒、抗增殖和免疫调节活性的细胞因子，可能影响库普弗细胞的活性。本研究旨在评估α干扰素或γ干扰素刺激后库普弗细胞的基因表达，以探讨这些细胞因子与巨噬细胞激活之间的联系。
将大鼠库普弗细胞培养24小时，分为三组：未刺激组；α干扰素刺激组；γ干扰素刺激组。刺激8小时后，按照Affymetrix方案提取并处理总RNA，然后与R34A微阵列基因集进行杂交。使用微阵列分析套件5.0软件进行数据分析。通过实时PCR对微阵列分析中显示出明显不同表达的基因进行确认。
阵列中代表的8800个基因中，近4000个基因在库普弗细胞中表达。其中，α干扰素使91个基因上调两倍以上（抗病毒、抗原加工和呈递以及肿瘤抑制/促凋亡基因），使72个基因下调50%或更多。γ干扰素使70个基因上调两倍以上，使78个基因下调50%或更多。α干扰素诱导的大多数基因也被γ干扰素诱导。下调的基因包括生长因子和参与细胞周期/增殖的基因。实时PCR证实了阵列的结果。
干扰素直接作用于大鼠库普弗细胞，参与多种基因的调控。它们的表达谱揭示了库普弗细胞在抗病毒和抗肿瘤等特定途径中激活的分子机制。
