| 王 璇,李学拥,李 靖,吕卓敏,何 林,芦希艳.Smurf1对增生性瘢痕纤维化形成的影响及分子机制[J].,2024,(5):849-857 |
| Smurf1对增生性瘢痕纤维化形成的影响及分子机制 |
| Effect of Smurf1 on Fibrosis Formation in Hypertrophic Scar and its Molecular Mechanism |
| 投稿时间：2023-11-20 修订日期：2023-12-15 |
| DOI：10.13241/j.cnki.pmb.2024.05.008 |
| 中文关键词: 增生性瘢痕 纤维化 人增生性瘢痕成纤维细胞 TGF-β1/Smad通路 增殖 侵袭 迁移 |
| 英文关键词: Hypertrophic scar Fibrosis Hypertrophic scar fibroblasts (HSF) TGF-β1/Smad pathway Proliferation Invasion Migration |
| 基金项目:陕西省重点研发计划项目（2023-YBSF-179） |
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| 中文摘要: |
| 摘要 目的：探究Smurf1对增生性瘢痕形成中纤维化进程的影响及分子机制。方法：收集2021年6月至2022年6月空军军医大学第二附属医院烧伤整形科行增生性瘢痕切除手术患者的瘢痕组织及正常皮肤标本各12例，采用HE和Masson染色进行病理学检查。取增生性瘢痕组织无菌处理后，采取组织块法分离培养获取人增生性瘢痕成纤维细胞（HSF）。将HSF细胞按照实验方案分组，（1）Smurf1过表达分组：对照1组（Con-1组），空载体组（Vector组）和Smurf1过表达组（OE-Smurf1组）；（2）Smurf1干扰表达分组：对照2组（Con-2组），阴性组（si-NC组），Smurf1干扰表达组（si-Smurf1组）。再分别将pcDNA3.1空质粒、pcDNA3.1+Smurf1质粒、si-NC和si-Smurf1转染至Vector组、OE-Smurf1组、si-NC组和si-Smurf1组HSF细胞。通过qRT-PCR检测Smurf1表达水平，Western blot检测蛋白表达水平，CCK-8检测细胞增殖水平，流式细胞术检测细胞凋亡水平，Transwell实验检测细胞侵袭水平，细胞划痕实验检测细胞迁移水平。结果：HE染色和Masson染色结果显示，与正常皮肤组织相比，增生性瘢痕组织的真皮层厚度显著增加，真皮层中存在大量被染成蓝色的胶原纤维，排列紊乱且致密。与正常皮肤组织相比，增生性瘢痕组织中TGF-β1、α-SMA、COL1、COL3、TβR-I、p-Smad3和Smad7的蛋白表达水平均升高（P<0.05），Smurf1表达水平降低（P<0.05）。与Con-1或Vector组比较，OE-Smurf1组HSF细胞中TGF-β1、α-SMA、COL1、COL3、TβR-I、p-Smad3和Smad7蛋白的表达水平均降低（P<0.05），HSF细胞增殖、侵袭和迁移水平降低（P<0.05），凋亡水平升高（P<0.05）。与Con-2组或si-NC组比较，si-Smurf1组HSF细胞中TGF-β1、α-SMA、COL1、COL3、TβR-I、p-Smad3和Smad7蛋白的表达水平均升高（P<0.05），HSF细胞增殖、侵袭和迁移水平升高（P<0.05），凋亡水平降低（P<0.05）。结论：Smurf1可能通过抑制TGF-β1/Smad通路，进而抑制增生性瘢痕的纤维化进程。 |
| 英文摘要: |
| ABSTRACT Objective: To explore the effect of Smurf1 on fibrosis process in hypertrophic scar formation and its molecular mechanism. Methods: Hypertrophic scar tissue and normal skin tissue samples of 12 patients who underwent hyperplastic scar resection in the department of burn and plastic surgery of the Second Affiliated Hospital of Air Force Medical University from June 2021 to June 2022 were collected. And the pathological examination was performed by HE and Masson staining. After sterile treatment of hypertrophic scar tissue, human hypertrophic scar fibroblasts (HSF) were isolated and cultured by tissue block method. HSF cells were divided into some groups according to the experimental scheme, (1) Smurf1 overexpression treatment group, control group 1 (Con-1 group), empty vector group (Vector group) and Smurf1 overexpression group (OE-Smurf1 group), (2) Smurf1 interference expression treatment group, control group 2 (Con-2 group), negative group (si-NC group) and Smurf1 interference expression group (si-Smurf1 group). Then pcDNA3.1 empty plasmid, pcDNA3.1+Smurf1 plasmid, si-NC and si-Smurf1 were transfected into HSF cells in Vector group, OE-Smurf1 group, si-NC group and si-Smurf1 group, respectively. Smurf1 mRNA expression level was detected by qRT-PCR. Protein expression level was detected by Western blot. Cell proliferation level was detected by CCK-8. Cell apoptosis level was detected by flow cytometry. Cell invasion level was detected by Transwell assay and cell migration level was detected by cell scratch assay. Results: The results of HE staining and Masson staining showed that compared with normal skin tissue, the dermis thickness of hypertrophic scar tissue increased significantly, and there were a large number of blue-dyed collagen fibers in the dermis, which were disordered and dense. Compared with normal skin tissue, the protein expression levels of TGF-β1, α-SMA, COL1, COL3, TβR-I, p-Smad3 and Smad7 in hypertrophic scar tissue were increased (P<0.05), while the expression level of Smurf1 was decreased (P<0.05). Compared with Con-1 or Vector groups, the expression levels of TGF-β1, α-SMA, COL1, COL3, TβR-I, p-Smad3 and Smad7 proteins in HSF cells of OE-Smurf1 group were decreased (P<0.05), and the proliferation, invasion and migration levels of HSF cells were decreased (P<0.05), the apoptosis level was increased (P<0.05). Compared with Con-2 group or si-NC group, the expression levels of TGF-β1, α-SMA, COL1, COL3, TβR-I, p-Smad3 and Smad7 proteins in HSF cells of si-Smurf1 group were increased (P<0.05), and the proliferation, invasion and migration levels of HSF cells were increased (P<0.05), the apoptosis level was decreased (P<0.05). Conclusion: Smurf1 may inhibit the fibrotic process of hypertrophic scars by inhibiting TGF-β1/Smad pathway. |
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