Human vascular smooth muscle cells. Target for and source of tumor necrosis factor.
Human vascular smooth muscle cells. Target for and source of tumor necrosis factor.
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DOI：
10.4049/jimmunol.142.1.100
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发表时间：
1989-01
影响因子：
4.4
通讯作者：
S. Warner;P. Libby
中科院分区：
文献类型：
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作者：
S. Warner;P. Libby
tnf - α（也被称为cachectin）可能通过作用于血管而在体内产生许多重要的作用。众所周知，内皮细胞对tnf - α有反应。我们研究了血管平滑肌细胞（SMC）是否也对tnf - α和相关细胞因子淋巴素（tnf - β）有反应。SMC是大多数血管中最丰富的细胞类型。人rtnf - α和β（0.1至100 ng/ml）均可诱导成人血管SMC瞬时积累IL-1 mRNA，并在1至4小时内达到峰值。RNA合成抑制剂放线菌素D（1微克/ml）可阻断IL-1 mRNA的诱导，而抑制蛋白质合成的环己亚胺（1微克/ml）可显著“超诱导”IL-1 α和IL-1 β mRNA。tnf - α处理也增加了细胞内生物活性IL-1和随后从SMC释放的IL-1活性。特异性抗体的代谢标记和免疫沉淀表明，在tnf处理或淋巴毒素处理的SMC中，IL-1 α和IL-1 β前体可以从头合成。tnf - α还激活了SMC的其他功能，包括PGE2从SMC的浓度依赖性释放，以及(2'-5')-寡聚腺苷酸合成酶基因的时间依赖性诱导，这种酶被认为介导IFN的抗病毒和抗增殖作用。我们还探讨了SMC是否也能表达TNF基因中的任何一种，SMC既能产生IL-1，又能对IL-1做出反应。细菌LPS（10微克/毫升）引起tnf - α转录物的轻微积累。SMC与蛋白质合成抑制剂单独孵育4小时，很少或没有引起tnf - α mRNA的升高，但同时添加LPS（“超诱导”条件）诱导了大量的tnf - α（但没有tnf - β） mRNA。细胞用大霉素（1微克/毫升）和LPS处理，然后洗涤以去除这种可逆的蛋白质合成抑制剂，释放tnf - α到培养基中，通过L929细胞毒性试验、代谢标记和免疫沉淀进行评估。因此，SMC对TNF和淋巴毒素都有反应，并能产生TNF- α， TNF- α是一种对血管细胞有多种影响的细胞因子，在感染性休克和其他炎症的病理生理中具有潜在的重要意义。
TNF-alpha (also known as cachectin) may produce many of its important effects in vivo by actions on blood vessels. Endothelial cells are well known to respond to TNF-alpha. We investigated whether vascular smooth muscle cells (SMC), the most abundant cell type in most vessels, also respond to TNF-alpha and the related cytokine lymphotoxin (TNF-beta). Both human rTNF-alpha and beta (0.1 to 100 ng/ml) induced transient accumulation of IL-1 mRNA by adult human vascular SMC that peaked between 1 and 4 h. The inhibitor of RNA synthesis actinomycin D (1 microgram/ml) blocked the induction of IL-1 mRNA, whereas inhibition of protein synthesis with cycloheximide (1 microgram/ml) resulted in a marked "superinduction" of both IL-1 alpha and IL-1 beta mRNA species. TNF-alpha treatment also increased intracellular biologically active IL-1 and subsequent release of IL-1 activity from SMC. Metabolic labeling and immunoprecipitation with specific antibodies demonstrated de novo synthesis of IL-1 alpha and IL-1 beta precursors in TNF-treated or lymphotoxin-treated SMC. TNF-alpha also activated other SMC functions including the concentration-dependent release of PGE2 from SMC, and time-dependent induction of the gene for (2'-5')-oligoadenylate synthetase, an enzyme thought to mediate the anti-viral and anti-proliferative actions of IFN. We also explored whether SMC, which both produce and respond to IL-1, might also express either of the TNF genes. Bacterial LPS (10 micrograms/ml) caused slight accumulation of TNF-alpha transcripts. Incubation of SMC for 4 h with inhibitors of protein synthesis alone caused little or no elevation of TNF-alpha mRNA, but simultaneous addition of LPS ("superinduction" conditions) induced large amounts of TNF-alpha (but not TNF-beta) mRNA. Cells treated with anisomycin (1 microgram/ml) and LPS, then washed to remove this reversible inhibitor of protein synthesis, released TNF-alpha into the medium, as assessed by the L929 cytotoxicity assay and by metabolic labeling and immunoprecipitation. Thus, SMC both respond to both TNF and lymphotoxin and can produce TNF-alpha, a cytokine with numerous effects on vascular cells of potential significance in the pathophysiology of septic shock and other inflammatory conditions.