Department of Plastic and Aesthetic Surgery, Peking Union Medical College Hospital, Peking Union Medical College & Chinese Academy of Medical Sciences, Beijing, China.
Beijing Key Laboratory (No.BZO381), Center of Excellence in Tissue Engineering Chinese Academy of Medical Sciences, School of Basic Medicine Peking Union Medical College, Institute of Basic Medical Sciences Chinese Academy of Medical Sciences, Beijing, China.
J Dermatol Sci. 2023 Oct;112(1):31-38. doi: 10.1016/j.jdermsci.2023.09.001. Epub 2023 Sep 6.
Inflammation and fibrosis of the skin are characteristics of localized scleroderma (LS). Emerging evidence has demonstrated that exosomes from human adipose tissue-derived mesenchymal stem cells (ADSC-Exo) could alleviate skin fibrosis.
The impact and potential mechanism of ADSC-Exo on LS fibrosis was examined.
ADSC-Exo was isolated and identified. The effects of ADSC-Exo on the abilities of proliferation and migration of LS-derived fibroblasts (LSFs) were assessed by CCK-8 and scratch assays, respectively. qRT-PCR, western blot, and immunofluorescence were conducted to detect LSFs stimulated with ADSC-Exo, ADSC-Exo, let-7a-5p mimic/TGF-βR1 shRNA virus, and negative controls. The impact of ADSC-Exo on C57BL/6j LS mice was evaluated by photographic morphology, hematoxylin-eosin (H&E), Masson's trichrome, and immunohistochemical staining.
The verified ADSC-Exo limited the proliferation and migration of LSFs and reduced the expression of COL1, COL3, α-SMA, TGF-βR1, and p-Smad2/ 3 in vitro and in vivo. TGF-βR1 knockdown and let-7a-5p mimic in LSFs reduced the expression of COL1, COL3, α-SMA, and p-Smad2/3. However, compared with the ADSC-Exo group, the dermal thickness was increased, collagen arrangement was disordered, and α-SMA and TGF-βR1 levels were increased after exposure to ADSC-Exo.
In this study, it might show that ADSC-Exo may successfully prevent LSF bioactivity, collagen deposition, and myofibroblast trans-differentiation. Additionally, we confirmed that let-7a-5p in ADSC-Exo could directly target TGF-R1 to control the Smad pathway and reduce fibrosis in LSFs. Our work offered a brand-new therapeutic approach and clarified the unique mechanism for the clinical management of LS.
皮肤炎症和纤维化是局限性硬皮病（LS）的特征。新出现的证据表明，人脂肪组织来源的间充质干细胞（ADSC-Exo）的外泌体可以减轻皮肤纤维化。
研究 ADSC-Exo 对 LS 纤维化的影响及其潜在机制。
分离并鉴定 ADSC-Exo。通过 CCK-8 和划痕实验分别评估 ADSC-Exo 对 LS 衍生成纤维细胞（LSFs）增殖和迁移能力的影响。qRT-PCR、western blot 和免疫荧光检测 LSFs 经 ADSC-Exo、ADSC-Exo、let-7a-5p 模拟物/TGF-βR1 shRNA 病毒和阴性对照刺激后的表达情况。通过摄影形态学、苏木精-伊红（H&E）、Masson 三色和免疫组织化学染色评估 ADSC-Exo 对 C57BL/6j LS 小鼠的影响。
经验证的 ADSC-Exo 限制了 LSFs 的增殖和迁移，并降低了 COL1、COL3、α-SMA、TGF-βR1 和 p-Smad2/3 在体外和体内的表达。LSFs 中的 TGF-βR1 敲低和 let-7a-5p 模拟物降低了 COL1、COL3、α-SMA 和 p-Smad2/3 的表达。然而，与 ADSC-Exo 组相比，暴露于 ADSC-Exo 后真皮厚度增加，胶原排列紊乱，α-SMA 和 TGF-βR1 水平升高。
在这项研究中，ADSC-Exo 可能成功地阻止了 LSF 的生物活性、胶原沉积和肌成纤维细胞转分化。此外，我们证实 ADSC-Exo 中的 let-7a-5p 可以直接靶向 TGF-R1 来控制 Smad 通路并减少 LSFs 中的纤维化。我们的工作提供了一种全新的治疗方法，并阐明了 LS 临床管理的独特机制。
