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与C57BL/6野生型小鼠相比,硫酸乙酰肝素缺陷的Hspg2外显子3缺失小鼠皮肤中TGF-β1的沉积减少。

The heparan sulphate deficient Hspg2 exon 3 null mouse displays reduced deposition of TGF-β1 in skin compared to C57BL/6 wild type mice.

作者信息

Shu Cindy, Smith Susan M, Melrose James

机构信息

Raymond Purves Bone and Joint Research Laboratory, Institute of Bone and Joint Research, Kolling Institute Northern Sydney Local Health District, The Royal North Shore Hospital, Level 10, Kolling Institute, Building B6, St. Leonards, NSW, 2065, Australia.

Sydney Medical School, Northern, The University of Sydney, Royal North Shore Hospital, St. Leonards, Australia.

出版信息

J Mol Histol. 2016 Jun;47(3):365-74. doi: 10.1007/s10735-016-9677-0. Epub 2016 Apr 21.

Abstract

This was an observational study where we examined the role of perlecan HS on the deposition of TGF-β1 in C57BL/6 and Hspg2(∆3-/∆3-) perlecan exon 3 null mouse skin. Despite its obvious importance in skin repair and tissue homeostasis no definitive studies have immunolocalised TGF-β1 in skin in WT or Hspg2(∆3-/∆3-) perlecan exon 3 null mice. Vertical parasagittal murine dorsal skin from 3, 6 and 12 week old C57BL/6 and Hspg2(∆3-/∆3-) mice were fixed in neutral buffered formalin, paraffin embedded and 4 μm sections stained with Mayers haematoxylin and eosin (H & E). TGF-β1 was immunolocalised using a rabbit polyclonal antibody, heat retrieval and the Envision NovaRED detection system. Immunolocalisation of TGF-β1 differed markedly in C57BL/6 and Hspg2(∆3-/∆3-) mouse skin, ablation of exon 3 of Hspg2 resulted in a very severe reduction in the deposition of TGF-β1 in skin 3-12 weeks postnatally. The reduced deposition of TGF-β1 observed in the present study would be expected to impact detrimentally on the remodelling and healing capacity of skin in mutant mice compounding on the poor wound-healing properties already reported for perlecan exon 3 null mice due to an inability to signal with FGF-2 and promote angiogenic repair processes. TGF-β1 also has cell mediated effects in tissue homeostasis and matrix stabilisation a reduction in TGF-β1 deposition would therefore be expected to detrimentally impact on skin homeostasis in the perlecan mutant mice.

摘要

这是一项观察性研究,我们研究了基底膜聚糖硫酸乙酰肝素(perlecan HS)在C57BL/6和基底膜聚糖基因(Hspg2)第3外显子缺失(Hspg2(∆3-/∆3-))小鼠皮肤中转化生长因子-β1(TGF-β1)沉积中的作用。尽管其在皮肤修复和组织稳态中具有明显重要性,但尚无明确研究对野生型(WT)或Hspg2(∆3-/∆3-)小鼠皮肤中的TGF-β1进行免疫定位。采集3周龄、6周龄和12周龄C57BL/6和Hspg2(∆3-/∆3-)小鼠的垂直矢状面背部皮肤,用中性缓冲福尔马林固定,石蜡包埋,制成4μm切片,用迈耶苏木精和伊红(H&E)染色。使用兔多克隆抗体、热修复和Envision NovaRED检测系统对TGF-β1进行免疫定位。TGF-β1在C57BL/6和Hspg2(∆3-/∆3-)小鼠皮肤中的免疫定位有显著差异,Hspg2第3外显子缺失导致出生后3至12周皮肤中TGF-β1的沉积严重减少。本研究中观察到的TGF-β1沉积减少预计会对突变小鼠皮肤的重塑和愈合能力产生不利影响,这使基底膜聚糖第3外显子缺失小鼠本已报道的伤口愈合不良情况更加严重,因为其无法与成纤维细胞生长因子-2(FGF-2)信号传导并促进血管生成修复过程。TGF-β 在组织稳态和基质稳定中也具有细胞介导作用,因此TGF-β1沉积减少预计会对基底膜聚糖突变小鼠的皮肤稳态产生不利影响。

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