Institute for Human Tissue Restoration, Department of Plastic & Reconstructive Surgery, Severance Hospital, Yonsei University College of Medicine, Seoul 03722, Korea.
College of Medicine, Graduate School, Yonsei University, Seoul 03722, Korea.
Int J Mol Sci. 2022 Jul 18;23(14):7918. doi: 10.3390/ijms23147918.
Wound healing is a complicated cascading process; disequilibrium among reparative processes leads to the formation of pathologic scars. Herein, we explored the role of mortalin in scar formation and its association with the interleukin-1α receptor using in vitro and in vivo models. To investigate the effects of mortalin, we performed an MTT cell viability assay, qRT-PCR, and Western blot analyses, in addition to immunofluorescence and immunoprecipitation studies using cultured fibroblasts. A rat incisional wound model was used to evaluate the effect of a mortalin-specific shRNA (dE1-RGD/GFP/shMot) Ad vector in scar tissue. In vitro, the mortalin-treated human dermal fibroblast displayed a significant increase in proliferation of type I collagen, α-smooth muscle actin, transforming growth factor-β, phospho-Smad2/3-complex, and NF-κB levels. Immunofluorescence staining revealed markedly increased mortalin and interleukin-1α receptor protein in keloid tissue compared to those in normal tissue, suggesting that the association between mortalin and IL-1α receptor was responsible for the fibrogenic effect. In vivo, mortalin-specific shRNA-expressing Ad vectors significantly decreased the scar size and type-I-collagen, α-SMA, and phospho-Smad2/3-complex expression in rat incisional scar tissue. Thus, dE1-RGD/GEP/shMot can inhibit the TGF-β/α-SMA axis and NF-κB signal pathways in scar formation, and blocking endogenous mortalin could be a potential therapeutic target for keloids.
伤口愈合是一个复杂的级联过程；修复过程的失衡会导致病理性瘢痕的形成。在此，我们通过体外和体内模型探索了 mortalin 在瘢痕形成中的作用及其与白细胞介素-1α 受体的关系。为了研究 mortalin 的作用，我们进行了 MTT 细胞活力测定、qRT-PCR 和 Western blot 分析，以及使用培养的成纤维细胞进行免疫荧光和免疫沉淀研究。采用大鼠切口伤口模型评估 mortalin 特异性 shRNA（dE1-RGD/GFP/shMot）Ad 载体在瘢痕组织中的作用。在体外，用 mortalin 处理的人真皮成纤维细胞显示 I 型胶原蛋白、α-平滑肌肌动蛋白、转化生长因子-β、磷酸化 Smad2/3 复合物和 NF-κB 水平的显著增殖。免疫荧光染色显示，与正常组织相比，瘢痕组织中的 mortalin 和白细胞介素-1α 受体蛋白明显增加，表明 mortalin 和 IL-1α 受体之间的关联是纤维生成效应的原因。在体内，mortalin 特异性 shRNA 表达的 Ad 载体显著减少了大鼠切口瘢痕组织中的瘢痕大小以及 I 型胶原蛋白、α-SMA 和磷酸化 Smad2/3 复合物的表达。因此，dE1-RGD/GEP/shMot 可以抑制 TGF-β/α-SMA 轴和 NF-κB 信号通路在瘢痕形成中的作用，阻断内源性 mortalin 可能是瘢痕疙瘩的潜在治疗靶点。
