Epidermal or dermal specific knockout of PHD-2 enhances wound healing and minimizes ischemic injury.
Epidermal or dermal specific knockout of PHD-2 enhances wound healing and minimizes ischemic injury.
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DOI：
10.1371/journal.pone.0093373
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Longaker MT
中科院分区：
文献类型：
--
作者：
Zimmermann AS;Morrison SD;Hu MS;Li S;Nauta A;Sorkin M;Meyer NP;Walmsley GG;Maan ZN;Chan DA;Gurtner GC;Giaccia AJ;Longaker MT
缺氧诱导因子 (HIF)-1α 是介导细胞对缺氧反应的异二聚体转录因子的一部分，对于多种血管生成生长因子的表达、细胞运动和内皮祖细胞的募集至关重要。抑制 HIF-1α 的氧依赖性负调节因子脯氨酰羟化酶结构域 2 (PHD-2)，会导致 HIF-1α 增加并模拟对缺氧的各种细胞和生理反应。 PHD-2 在表皮和真皮中在伤口愈合中的作用尚未明确。通过将纯合 floxed PHD-2 小鼠与杂合 K14-Cre 小鼠和杂合 Col1A2-Cre-ER 小鼠杂交，在 C57BL/6J（野生型）背景下培育表皮和真皮特异性 PHD-2 敲除（KO）小鼠，以获得纯合 floxed PHD-2/杂合 K14-Cre 和纯合 floxed 分别为 PHD-2/杂合 floxed Col1A2-Cre-ER 小鼠。对 10 至 12 周大的 PHD-2 KO 和野生型 (WT) 小鼠进行创伤和缺血蒂皮瓣模型。使用 ImageJ 软件对愈合量进行了粗略量化。对体外培养的原代细胞的蛋白质和 RNA 进行蛋白质印迹和 qRT-PCR。 qRT-PCR 证明，相对于对照小鼠，来自组织特异性 KO 小鼠的角质形成细胞和成纤维细胞中的 PHD-2 显着减少 (*p<0.05)。 Western blot 分析显示，相对于对照小鼠，PHD-2 KO 小鼠的 HIF-1α 和 VEGF 蛋白水平显着增加 (*p<0.05)。相对于 WT 小鼠，PHD-2 KO 小鼠的伤口闭合速度显着加快（*p<0.05）。当在皮瓣模型中进行术后第九天的缺血分析时，PHD-2组织特异性敲除小鼠显示出比WT小鼠明显更多的存活皮瓣（*p<0.05）。 PHD-2 在小鼠伤口愈合速度和对缺血性损伤的反应中发挥着重要作用。进一步的研究表明，PHD-2 KO 会增加 HIF-1α 的细胞水平，这种增加会导致下游血管生成因子（如 VEGF）的转录。
Hypoxia-inducible factor (HIF)-1α, part of the heterodimeric transcription factor that mediates the cellular response to hypoxia, is critical for the expression of multiple angiogenic growth factors, cell motility, and the recruitment of endothelial progenitor cells. Inhibition of the oxygen-dependent negative regulator of HIF-1α, prolyl hydroxylase domain-2 (PHD-2), leads to increased HIF-1α and mimics various cellular and physiological responses to hypoxia. The roles of PHD-2 in the epidermis and dermis have not been clearly defined in wound healing. Epidermal and dermal specific PHD-2 knockout (KO) mice were developed in a C57BL/6J (wild type) background by crossing homozygous floxed PHD-2 mice with heterozygous K14-Cre mice and heterozygous Col1A2-Cre-ER mice to get homozygous floxed PHD-2/heterozygous K14-Cre and homozygous floxed PHD-2/heterozygous floxed Col1A2-Cre-ER mice, respectively. Ten to twelve-week-old PHD-2 KO and wild type (WT) mice were subjected to wounding and ischemic pedicle flap model. The amount of healing was grossly quantified with ImageJ software. Western blot and qRT-PCR was run on protein and RNA from primary cells cultured in vitro. qRT-PCR demonstrated a significant decrease of PHD-2 in keratinocytes and fibroblasts derived from tissue specific KO mice relative to control mice (*p<0.05). Western blot analysis showed a significant increase in HIF-1α and VEGF protein levels in PHD-2 KO mice relative to control mice (*p<0.05). PHD-2 KO mice showed significantly accelerated wound closure relative to WT (*p<0.05). When ischemia was analyzed at day nine post-surgery in a flap model, the PHD-2 tissue specific knockout mice showed significantly more viable flaps than WT (*p<0.05). PHD-2 plays a significant role in the rates of wound healing and response to ischemic insult in mice. Further exploration shows PHD-2 KO increases cellular levels of HIF-1α and this increase leads to the transcription of downstream angiogenic factors such as VEGF.
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影响因子：
3.1
作者：
Cross, KJ;Mustoe, TA
通讯作者：
Mustoe, TA
DOI：
10.1073/pnas.0906670106
发表时间：
2009-08-11
影响因子：
11.1
作者：
Thangarajah, Hariharan;Yao, Dachun;Gurtner, Geoffrey C.
通讯作者：
Gurtner, Geoffrey C.
影响因子：
4.3
作者：
Driver, Vickie R.;Fabbi, Matteo;Gibbons, Gary
通讯作者：
Gibbons, Gary
影响因子：
82.9
作者：
Ceradini, DJ;Kulkarni, AR;Gurtner, GC
通讯作者：
Gurtner, GC
影响因子：
6
作者：
Zheng, B;Zhang, ZP;Denton, CP
通讯作者：
Denton, CP