Kameyama Hiroshi, Udagawa Orie, Hoshi Tomoaki, Toukairin Yoko, Arai Tomomi, Nogami Makoto
Criminal Investigation Laboratory, Saitama Prefectural Police Headquarters, 3-15-1, Takasago, Urawa-ku, Saitama City, Saitama 330-8533, Japan; Department of Legal Medicine, Teikyo University School of Medicine, 2-11-1, Kaga, Itabashi-ku, Tokyo 173-8605, Japan.
Department of Legal Medicine, Teikyo University School of Medicine, 2-11-1, Kaga, Itabashi-ku, Tokyo 173-8605, Japan.
Leg Med (Tokyo). 2015 Jul;17(4):255-60. doi: 10.1016/j.legalmed.2015.02.007. Epub 2015 Mar 7.
Wound healing evaluation is important in forensic pathology, in which angiogenesis plays an important role. We have already shown that vascular endothelial growth factor A (VEGF) is produced in the rat skin incision wounds by neutrophils, endothelial cells, and fibroblasts. In this study, we assessed the changes in the mRNA expressions of various factors possibly involved in angiogenesis including angiopoietin (ANGPT) 1 and 2, cadherin 5 (CDH5), granulocyte-macrophage colony stimulating factor (CSF2/GM-CSF), granulocyte colony stimulating factor (CSF3/G-CSF), chemokine (C-X-C motif) ligand 2 (CXCL2), chemokine (C-X-C motif) ligand12 (CXCL12/SDF1), endothelin 1 (ET1), fibroblast growth factor 1 (FGF 1), hepatocyte growth factor (HGF), hypoxia inducible factor 1 alpha (HIF1a), leptin, matrix metallopepitidase 9 (MMP9), serpine/plasminogen activator inhibitor1 (PAI1), platelet-derived growth factor-A (PDGF-A), transforming growth factor alpha and beta 1 (TGFa and b1), tenomodulin (TNMD), and troponin I type 2 (TNNI2) in the early stage of the rat skin incision wounds by real time RT-PCR. Factors reported to be involved in lymphangiogenesis such as fibroblast growth factor 2 (FGF 2), c-fos induced growth factor (FIGF/VEGF-D), forkhead box C2 (FOXC2), and prospero homeobox 1 (PROX1) were also studied. One and 3 days after the dorsal skin incisions, wounds on male Sprague-Dawley rats showed the statistically significant increases in the mRNA expressions for CXCL2, CSF3, MMP9, PAI1, and CSF2, whereas TGFa, TNNI2, FGF1, TNMD, leptin, and CXCL12 showed the statistically significant decreases. Interestingly, lymphgangiogenic factors FOXC2, PROX1, and FGF2 also showed the statistically significant decreases. In situ hybridization and immunohistochemistry showed the mRNA and protein positivity in endothelial cells, fibroblasts, and some leukocytes at the bottom of the wound tissue for PAI1, CSF3, and MMP9, 1 day after the skin incisions. Our novel findings show the possible involvement of several factors involved in angiogenesis and lymphangiogenesis in the early stage of wound healing process, which may be useful for forensic wound evaluations.
伤口愈合评估在法医病理学中很重要，其中血管生成起着重要作用。我们已经表明，中性粒细胞、内皮细胞和成纤维细胞在大鼠皮肤切口伤口中产生血管内皮生长因子A（VEGF）。在本研究中，我们通过实时RT-PCR评估了大鼠皮肤切口伤口早期可能参与血管生成的各种因子的mRNA表达变化，这些因子包括血管生成素（ANGPT）1和2、钙黏蛋白5（CDH5）、粒细胞-巨噬细胞集落刺激因子（CSF2/GM-CSF）、粒细胞集落刺激因子（CSF3/G-CSF）、趋化因子（C-X-C基序）配体2（CXCL2）、趋化因子（C-X-C基序）配体12（CXCL12/SDF1）、内皮素1（ET1）、成纤维细胞生长因子1（FGF 1）、肝细胞生长因子（HGF）、缺氧诱导因子1α（HIF1a）、瘦素、基质金属肽酶9（MMP9）、丝氨酸蛋白酶/纤溶酶原激活物抑制剂1（PAI1）、血小板衍生生长因子-A（PDGF-A）、转化生长因子α和β1（TGFα和β1）、肌腱调节蛋白（TNMD）以及肌钙蛋白I 2型（TNNI2）。还研究了据报道参与淋巴管生成的因子，如成纤维细胞生长因子2（FGF 2）、c-fos诱导生长因子（FIGF/VEGF-D）、叉头框C2（FOXC2）和prospero同源盒1（PROX1）。在背部皮肤切开1天和3天后，雄性Sprague-Dawley大鼠伤口处CXCL2、CSF3、MMP9、PAI1和CSF2的mRNA表达有统计学意义的增加，而TGFα、TNNI2、FGF1、TNMD、瘦素和CXCL12有统计学意义的下降。有趣的是，淋巴管生成因子FOXC2、PROX1和FGF2也有统计学意义的下降。原位杂交和免疫组化显示，皮肤切开1天后，伤口组织底部的内皮细胞、成纤维细胞和一些白细胞中PAI1、CSF3和MMP9的mRNA和蛋白呈阳性。我们的新发现表明，在伤口愈合过程的早期，几种参与血管生成和淋巴管生成的因子可能发挥作用，这可能对法医伤口评估有用。
