Mauviel A, Temime N, Charron D, Loyau G, Pujol J P
Laboratoire de Biochimie du Tissu Conjonctif, C.H.U. Côte de Nacre, Caen, France.
J Cell Biochem. 1991 Oct;47(2):174-83. doi: 10.1002/jcb.240470211.
It has previously been demonstrated that interleukin-1 (IL-1) is expressed in a variety of fibroblast cell lines. In this study, we investigated the mechanisms involved in the regulation of IL-1 beta production by cultured human dermal fibroblasts. We have shown that IL-1 beta is constitutively expressed as a cell-associated form, with no soluble form detectable in control cell or in stimulated cell supernatants. IL-1 alpha and tumor necrosis factor-alpha (TNF-alpha) exerted a dose-dependent stimulation on the production of the cell-associated IL-1 beta, as estimated using a specific enzyme linked immunosorbent assay (ELISA). As expected, this effect was accompanied by a huge release of prostaglandin E2 (PGE2) and a transient rise in intracellular cyclic AMP. Furthermore, IL-1 beta production was elevated to a lesser extent by the addition of increasing concentrations of the protein kinase C activator phorbol myristate acetate or by low concentration (0.001 microgram/ml) of PGE2. In contrast, higher concentrations (0.1 and 1 micrograms/ml) of PGE2, as well as exogenous dibutyryl-cyclic AMP, were clearly inhibitory. H7, an inhibitor of protein kinases also reduced the stimulatory effect of IL-1 alpha and TNF-alpha. Together with the results obtained with phorbol myristate acetate, these data suggest that protein kinase C may play a role in the upregulation of IL-1 beta expression in normal skin fibroblasts. The addition of indomethacin not only suppressed prostaglandin synthesis, but also dramatically reduced cyclic AMP formation, probably because the PGE2-induced stimulation of adenylate cyclase was abolished. This resulted in a strong potentiation of the stimulatory effect of IL-1 alpha and TNF-alpha, supporting the role of both the cyclooxygenase and adenylate cyclase pathways in the endogenous downregulation of IL-1 beta induction by the two cytokines studied.
先前已证明白细胞介素-1（IL-1）在多种成纤维细胞系中表达。在本研究中，我们调查了培养的人皮肤成纤维细胞中IL-1β产生的调控机制。我们已表明，IL-1β以细胞相关形式组成性表达，在对照细胞或刺激细胞的上清液中未检测到可溶性形式。使用特异性酶联免疫吸附测定（ELISA）估计，IL-1α和肿瘤坏死因子-α（TNF-α）对细胞相关IL-1β的产生具有剂量依赖性刺激作用。正如预期的那样，这种作用伴随着前列腺素E2（PGE2）的大量释放和细胞内环磷酸腺苷的短暂升高。此外，添加浓度不断增加的蛋白激酶C激活剂佛波醇肉豆蔻酸酯乙酸盐或低浓度（0.001微克/毫升）的PGE2可使IL-1β的产生在较小程度上升高。相比之下，较高浓度（0.1和1微克/毫升）的PGE2以及外源性二丁酰环磷酸腺苷具有明显的抑制作用。蛋白激酶抑制剂H7也降低了IL-1α和TNF-α的刺激作用。连同用佛波醇肉豆蔻酸酯乙酸盐获得的结果，这些数据表明蛋白激酶C可能在正常皮肤成纤维细胞中IL-1β表达的上调中起作用。吲哚美辛的添加不仅抑制了前列腺素的合成，而且还显著降低了环磷酸腺苷的形成，这可能是因为PGE2诱导的腺苷酸环化酶刺激被消除了。这导致IL-1α和TNF-α的刺激作用强烈增强，支持了环氧合酶和腺苷酸环化酶途径在这两种研究的细胞因子对内源性IL-1β诱导的下调中的作用。
