WOUND MACROPHAGES EXPRESS TGF-ALPHA AND OTHER GROWTH-FACTORS INVIVO - ANALYSIS BY MESSENGER-RNA PHENOTYPING
WOUND MACROPHAGES EXPRESS TGF-ALPHA AND OTHER GROWTH-FACTORS INVIVO - ANALYSIS BY MESSENGER-RNA PHENOTYPING
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DOI：
10.1126/science.3041594
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发表时间：
1988-08-05
期刊：
影响因子：
56.9
通讯作者：
WERB, Z
中科院分区：
文献类型：
--
作者：
RAPPOLEE, DA;MARK, D;WERB, Z
巨噬细胞的存在是伤口愈合过程中许多细胞类型再生所必需的。据报道，巨噬细胞表达广泛的促有丝分裂因子和细胞因子，但这些因子中没有一个在体内显示出维持所有的伤口愈合过程。已经提出转化因子-α.（TGF-α）可以介导体内血管生成、表皮再生和肉芽组织的形成。从伤口部位分离的巨噬细胞，并且未暴露于细胞培养条件，表达TGF-α的信使RNA转录物，TGF-β，血小板衍生生长因子A链和胰岛素样生长因子-1。这些成绩单的表达确定了一种新的方法，RNA分析，其中低数量的小鼠巨噬细胞从伤口圆柱体中分离出来，他们的RNA纯化和逆转录，并在聚合酶链反应与生长因子序列特异性引物引物扩增的互补DNA。这种单细胞RNA表型分析方法快速，具有定量的潜力，可以明确地证明来自单个细胞或几个细胞的mRNA转录本，同时分析几种mRNA。来自伤口的巨噬细胞表达TGF-α。抗原，并且伤口流体含有TGF-α。培养物中的诱发巨噬细胞也表达TGF-α。在用修饰的低密度脂蛋白和脂多糖内毒素刺激后，转录物和多肽以时间依赖性方式表达，这是在损伤组织中发现的激活剂的特征。
The presence of macrophages is required for the regeneration of many cell types during wound healing. Marcrophages have been reported to express a wide range of mitogenic factors and cytokines, but none of these factors has shown in vivo to sustain all the wound-healing process. It has been suggested that transforming factor-.alpha. (TGF-.alpha.) may mediate angiogenesis, epidermal regrowth, and formation of granulation tissue in vivo. Macrophages isolated from a wound site, and not exposed to cell culture conditions, expressed messenger RNA transcript fro TGF-.alpha., TGF-.beta., platelet-derived growth factor A-chain, and insulin-like growth factor-1. The expression of these transcripts was determined by a novel method for RNA analysis in which low numbers of mouse macrophages were isolated from wound cylinders, their RNA was purified and reverse-transcribed, and the complementary DNA was amplified in a polymerase chain reaction primed with growth factor sequence-specific primers. This single-cell RNA phenotyping procedure is rapid and has the potential for quantification, and mRNA transcripts from a single cell or a few cells can be unambiguously demonstrated, with the simultaneous analysis of several mRNA species. Macrophages from wounds expressed TGF-.alpha. antigen, and wound fluids contained TGF-.alpha.. Elicited macrophages in culture also expressed TGF-.alpha. transcripts and polypeptide in a time-dependent manner after stimulation with modified low-density lipoproteins and lipopolysaccharide endotoxin, which are characteristic of the activators found in injured tissues.