Wound Macrophages Express TGF-α and Other Growth Factors in Vivo: Analysis by mRNA Phenotyping

伤口愈合 生物 肉芽组织 生长因子 核糖核酸 信使核糖核酸 分子生物学 表皮生长因子 体内 转化生长因子 细胞生物学 转化生长因子-α 细胞生长 细胞培养 基因 免疫学 生物化学 生物技术 受体 遗传学
作者
Daniel A. Rappolee,Mark David,Michael J. Banda,Zena Werb
出处
期刊:Science [American Association for the Advancement of Science]
卷期号:241 (4866): 708-712 被引量:1110
标识
DOI:10.1126/science.3041594
摘要

The presence of macrophages is required for the regeneration of many cell types during wound healing. Macrophages have been reported to express a wide range of mitogenic factors and cytokines, but none of these factors has been shown in vivo to sustain all the wound-healing processes. It has been suggested that transforming growth factor-α (TGF-α) 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 transcripts for TGF-α, TGF-β, 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-α antigen, and wound fluids contained TGF-α. Elicited macrophages in culture also expressed TGF-α 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.
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