Pig MAP/ITIH4 and haptoglobin are interleukin-6-dependent acute-phase plasma proteins in porcine primary cultured hepatocytes

被引:61
作者
González-Ramón, N
Hoebe, K
Alava, MA
van Leengoed, L
Piñeiro, M
Carmona, S
Iturralde, M
Lampreave, F
Piñeiro, A
机构
[1] Univ Zaragoza, Dept Bioquim & Biol Mol & Celular, E-50009 Zaragoza, Spain
[2] DLO, Inst Anim Sci & Hlth, Dept Pathobiol & Epidemiol, Lelystad, Netherlands
[3] Univ Utrecht, Dept Herd Hlth & Reprod, NL-3508 TC Utrecht, Netherlands
来源
EUROPEAN JOURNAL OF BIOCHEMISTRY | 2000年 / 267卷 / 06期
关键词
acute-phase proteins; cytokines; hepatocytes; inter-alpha-trypsin inhibitor (I alpha I); ITIH4;
D O I
10.1046/j.1432-1327.2000.01195.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The acute-phase expression of pig MAP (major acute-phase protein)/ITIH4 (inter-alpha-trypsin inhibitor heavy chain 4) and haptoglobin were analysed in primary cultures of isolated pig hepatocytes in response to recombinant human (rh) cytokines: tumor necrosis factor-alpha (TNF-alpha), interleukin-1 (IL-1), interleukin-6 (IL-6), as well as to bacterial lipopolysaccharide (LPS). Analysis of pig MAP/ITIH4 and haptoglobin mRNAs was carried out by RT-PCR amplification. Secreted proteins from the cytokine-treated hepatocytes were quantified by immunochemical techniques. Time-course and dose-response experiments show that pig MAP/ITIH4 and haptoglobin belong to the type II acute-phase proteins, as they are specifically induced by rhIL-6 and not by rhTNF-alpha or rhIL-1. Stimulation of cultured pig hepatocytes with rhIL-6 for 48 h at doses of 1000 U.mL(-1) showed a fourfold to fivefold increase in pig MAP/ITIH4 concentration in the medium, while the concentration of haptoglobin only increased twofold. A similar increase in the concentration of pig MAP/ITIH4 was also observed in media of LPS-treated hepatocytes with the simultaneous generation of IL-6 by the Kupfer cells present in the cultures. Albumin secretion decreased after stimulation with doses of 100 or 1000 U.mL(-1) rhTNF-alpha, rhIL-1 or rhIL-6. Therefore, it can be concluded that pig MAP/ITIH4 behaves as a major acute-phase protein produced by porcine hepatocytes under the effect of inflammatory cytokines.
引用
收藏
页码:1878 / 1885
页数:8
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