Han J, Thompson P, Beutler B
Howard Hughes Medical Institute, University of Texas, Southwestern Medical Center, Dallas 75235.
J Exp Med. 1990 Jul 1;172(1):391-4. doi: 10.1084/jem.172.1.391.
The induction of cachectin/tumor necrosis factor (TNF) synthesis by bacterial endotoxins is a process that entails activation at several levels. Cachectin/TNF gene transcription is accelerated, leading to rapid accumulation of mRNA within the macrophage cytosol. In addition, translational derepression occurs, leading to far more efficient message utilization. Through the use of posttranscriptional reporter constructs, we now demonstrate that certain agents capable of inhibiting cachectin/TNF biosynthesis operate through different mechanisms. In RAW 264.7 macrophages, pentoxifylline blocks cachectin/TNF mRNA accumulation but has no effect upon the efficiency of reporter mRNA translation. Dexamethasone, on the other hand, has only a modest effect on cachectin/TNF mRNA accumulation, but strongly impedes translational derepression. Combined application of dexamethasone and pentoxifylline to macrophages causes a greater suppression of cachectin/TNF biosynthesis that can be achieved by either agent alone. These findings suggest that the signaling pathway activated by endotoxin is branched, and that selective inhibition of different parts of the pathway may be achieved through the use of distinct agents.
细菌内毒素诱导恶病质素/肿瘤坏死因子(TNF)合成是一个涉及多个水平激活的过程。恶病质素/TNF基因转录加速,导致巨噬细胞胞质溶胶内mRNA迅速积累。此外,发生翻译去阻遏,导致更有效的信息利用。通过使用转录后报告构建体,我们现在证明某些能够抑制恶病质素/TNF生物合成的试剂通过不同机制起作用。在RAW 264.7巨噬细胞中,己酮可可碱阻断恶病质素/TNF mRNA积累,但对报告mRNA翻译效率没有影响。另一方面,地塞米松对恶病质素/TNF mRNA积累只有适度影响,但强烈阻碍翻译去阻遏。将地塞米松和己酮可可碱联合应用于巨噬细胞会导致对恶病质素/TNF生物合成的抑制作用比单独使用任何一种试剂都更大。这些发现表明内毒素激活的信号通路是分支的,并且通过使用不同的试剂可以实现对该通路不同部分的选择性抑制。