Importance of Housekeeping Gene Optimization for the Analysis of mRNA Expression During Wound Healing in a Third-Degree Burn Injury Model
Importance of Housekeeping Gene Optimization for the Analysis of mRNA Expression During Wound Healing in a Third-Degree Burn Injury Model
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管家基因优化对于三度烧伤模型伤口愈合过程中 mRNA 表达分析的重要性
DOI：
10.1093/jbcr/irac161
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发表时间：
2022
期刊：
影响因子：
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通讯作者：
Yoshida Toshiko
中科院分区：
文献类型：
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作者：
Arai Kenichi;Okabe Motonori;Kobashi Daisuke;Ichimura Kenji;Fathy Moustafa;Oba Jiro;Furuichi Etsuko;Yoshida Satoshi;Yoshida Toshiko
三度烧伤模型中的伤口愈合评价方法分为组织学（上皮再生和肉芽组织形成）和分子学（定量聚合酶链反应）。一般来说，mRNA表达相对于管家基因的表达进行标准化。虽然管家基因的表达通常是稳定的，但据报道，这些基因的稳定性取决于伤口愈合过程和治疗方法。在这项研究中，我们确定了最稳定的管家基因（TATA结合蛋白），用于研究三度烧伤模型中的基因表达，在该模型中，通过移植人骨髓间充质细胞促进伤口愈合。我们研究了人骨髓间充质细胞在损伤模型中的伤口愈合作用。肉芽组织的形成，从成纤维细胞分化成肌成纤维细胞，和功能性血管结构，促进在全层皮肤切除部位与这些细胞的治疗。测量血管生成、促炎和抗炎相关mRNA的表达，并将其标准化为管家基因的表达，表明用细胞处理促进了内皮细胞的浸润和M1和M2巨噬细胞的分化。总之，可以使用优化的管家基因准确分析三度烧伤模型中的伤口愈合。
Wound healing evaluation methods in a third-degree burn injury model are categorized as histological (re-epithelialization and granulation tissue formation) and molecular (quantitative polymerase chain reaction). In general, mRNA expression is normalized to those of the housekeeping gene. Although the housekeeping gene expression is generally stable, it has been reported that the stability of these genes depends on the wound healing process and treatment method. In this study, we identified the most stable housekeeping gene (TATA-binding protein) for studying gene expression in a third-degree burn injury model, in which wound healing was promoted by grafting human amnion-derived mesenchymal cells. We investigated the wound healing effect of human amnion-derived mesenchymal cells in the injury model. The formation of granulation tissue, the differentiation from fibroblasts to myofibroblasts, and functional vascular structure were promoted in the full-thickness skin excision site by treatment with these cells. The expression of angiogenic, pro-inflammatory and anti-inflammatory related mRNA was measured and normalized to that of the housekeeping gene, showing that treatment with the cells promoted the infiltration of endothelial cells and differentiation of M1 and M2 macrophages. In conclusion, wound healing in a third-degree burn injury model can be accurately analyzed using the optimized housekeeping gene.