基于“细菌超载”证据的增生性瘢痕治疗新方式探索

doi:10.3969/j.issn.1674-8115.2026.05.004

上海交通大学学报（医学版） ›› 2026, Vol. 46 ›› Issue (5): 585-593.doi: 10.3969/j.issn.1674-8115.2026.05.004

• 论著 · 基础研究 • 上一篇

基于“细菌超载”证据的增生性瘢痕治疗新方式探索

原博¹, 余佳容¹, 张铮², 王西樵¹(), 刘琰¹, 夏照帆³

^1.上海交通大学医学院附属瑞金医院烧伤整形与创面修复科，上海 200025
^2.上海交通大学医学院附属第九人民医院整复外科，上海 200011
^3.海军军医大学第一附属医院烧伤外科，上海 200433

收稿日期:2025-11-19 接受日期:2026-01-19 出版日期:2026-05-28 发布日期:2026-05-28
通讯作者: 王西樵，副研究员，博士；电子信箱：wxqiao2002@hotmail.com。
基金资助:
国家自然科学基金(82472555，82472544);上海市重中之重研究中心建设项目(2023ZZ02013)

Novel approach to hypertrophic scar treatment based on evidence of bacterial overload

Yuan Bo¹, Yu Jiarong¹, Zhang Zheng², Wang Xiqiao¹(), Liu Yan¹, Xia Zhaofan³

^1.Department of Burns, Plastic Surgery and Wound Repair, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
^2.Department of Plastic and Reconstructive Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China
^3.Department of Burn Surgery, The First Affiliated Hospital of Naval Medical University, Shanghai 200433, China

Received:2025-11-19 Accepted:2026-01-19 Online:2026-05-28 Published:2026-05-28
Contact: Wang Xiqiao, E-mail: wxqiao2002@hotmail.com.
Supported by:
National Natural Science Foundation of China(82472555，82472544);Shanghai Top Priority Research Center Project(2023ZZ02013)

摘要/Abstract

摘要：

目的·观察体外细菌对成纤维细胞功能的影响，以及体内细菌干预对兔耳增生性瘢痕的影响。方法·收集2023年1月——2024年12月上海交通大学医学院附属瑞金医院烧伤整形与创面修复科的16例临床增生性瘢痕标本和正常皮肤组织，进行革兰染色。标准金黄色葡萄球菌扩增灭活后，以10²、10³和10⁴CFU/mL干预正常皮肤来源的成纤维细胞。采用Edu染色观察金黄色葡萄球菌对成纤维细胞增殖的影响，免疫荧光染色法检测α-平滑肌肌动蛋白（α-smooth muscle actin，α-SMA）表达，Western blotting检测血管内皮生长因子（vascular permeability factor，VEGF）、转化生长因子-β1（transforming growth factor-β1，TGF-β1）、Ⅰ型胶原及Ⅲ型胶原蛋白表达，酶联免疫吸附试验（enzyme linked immunosorbent assay，ELISA）检测金黄色葡萄球菌对白细胞介素-6（interleukin-6，IL-6）、IL-8、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）及单核细胞趋化蛋白-1（monocyte chemoattractant protein-1，MCP-1）等炎症因子分泌的影响。选取10只新西兰白兔，双耳共建立80个创面，创面采用金黄色葡萄球菌菌液处理，待增生性瘢痕形成后，予瘢痕皮内注射左氧氟沙星或生理盐水，观察瘢痕生长情况并进行温哥华瘢痕量表评分。兔耳瘢痕取材后，进行苏木精-伊红（hematoxylin-eosin，HE）染色、Masson染色和革兰染色，采用免疫组织化学检测IL-6、IL-8、TNF-α及MCP-1等炎症因子的表达。结果·临床瘢痕组织内存在大量革兰染色阳性菌，与正常皮肤比较差异有统计学意义（P=0.001）。体外实验证明：低丰度细菌（10²、10³ CFU/mL）可显著促进成纤维细胞增殖（P=0.045，P=0.017）和α-SMA表达（P=0.042，P=0.002），促进VEGF（P=0.023，P=0.011）、TGF-β1（P=0.029，P=0.031）、Ⅰ型胶原蛋白（P=0.032，P=0.025）、Ⅲ型胶原蛋白表达（P=0.019，P=0.027），以及促进炎症因子IL-6（P=0.023，P=0.011）、IL-8（P=0.021，P=0.009）、TNF-α（P=0.029，P=0.011）及MCP-1（P=0.023，P=0.008）分泌。兔耳瘢痕中注射左氧氟沙星后，45 d和60 d时瘢痕增生减轻（P=0.019，P=0.013）；60 d时瘢痕炎症细胞减少，细菌含量降低，胶原蛋白沉积变少，炎症因子IL-6（P=0.025）、IL-8（P=0.021）、MCP-1（P=0.028）及TNF-α（P=0.019）等表达下降。结论·低丰度细菌促进瘢痕成纤维细胞致纤维化能力，抗生素应用可有效减轻增生性瘢痕的形成。

关键词: 增生性瘢痕, 细菌, 成纤维细胞, 炎症因子

Abstract:

Objective ·To investigate the effects of bacteria on fibroblast function in vitro and the impact of bacterial intervention on hypertrophic scars in rabbit ears in vivo. Methods ·A total of 16 clinical hypertrophic scar specimens and normal skin tissues were collected from patients admitted to the Department of Burns, Plastic Surgery and Wound Repair, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, from January 2023 to December 2024, and subjected to Gram staining. After amplification and inactivation, standard Staphylococcus aureus was used to treat fibroblasts derived from normal skin at concentrations of 10², 10³, and 10⁴ CFU/mL. The effects on fibroblast proliferation were observed by using Edu staining, while α-smooth muscle actin (α-SMA) expression was detected by immunofluorescence. The expression levels of vascular endothelial growth factor (VEGF), transforming growth factor-β1 (TGF-β1), type Ⅰ collagen and type Ⅲ collagen were measured by Western blotting. Additionally, the secretion of inflammatory cytokines, including interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α), and monocyte chemoattractant protein-1 (MCP-1), was evaluated by using enzyme-linked immunosorbent assay (ELISA). In the in vivo study, 80 wounds were created on the ears of 10 New Zealand white rabbits, followed by inoculation with Staphylococcus aureus. After hypertrophic scar formation, intradermal injections of levofloxacin or saline were administered. Scar growth was monitored, and Vancouver Scar Scale scores were recorded. Scar tissues were harvested for hematoxylin and eosin staining, Masson staining, and Gram staining, along with immunohistochemical detection of IL-6, IL-8, TNF-α, and MCP-1 expression. Results ·Clinical scar tissues exhibited significantly greater abundance of Gram-positive bacteria than normal skin tissues (P=0.001). In vitro experiments demonstrated that low-abundance bacteria (10² and 10³ CFU/mL) significantly promoted fibroblast proliferation (P=0.045, P=0.017) and α-SMA expression (P=0.042, P=0.002). These bacteria also enhanced the expression of VEGF (P=0.023, P=0.011), TGF-β1 (P=0.029, P=0.031), type Ⅰ collagen (P=0.032, P=0.025), and type Ⅲ collagen (P=0.019, P=0.027), as well as the secretion of inflammatory cytokines IL-6 (P=0.023, P=0.011), IL-8 (P=0.021, P=0.009), TNF-α (P=0.029, P=0.011), and MCP-1 (P=0.023, P=0.008). In the rabbit ear model, levofloxacin injection significantly reduced scar hyperplasia at 45 and 60 days (P=0.019, P=0.013). At 60 days, treated scars showed decreased inflammatory cell infiltration, reduced bacterial load, less collagen deposition, and lower expressions of IL-6 (P=0.025), IL-8 (P=0.021), MCP-1 (P=0.028), and TNF-α (P=0.019). Conclusions ·Low-abundance bacteria promote the profibrotic capacity of scar fibroblasts, while antibiotic application effectively mitigates hypertrophic scar formation.

Key words: hypertrophic scar, bacteria, fibroblast, inflammatory cytokine

中图分类号:

R453.2

原博, 余佳容, 张铮, 王西樵, 刘琰, 夏照帆. 基于“细菌超载”证据的增生性瘢痕治疗新方式探索[J]. 上海交通大学学报（医学版）, 2026, 46(5): 585-593.

Yuan Bo, Yu Jiarong, Zhang Zheng, Wang Xiqiao, Liu Yan, Xia Zhaofan. Novel approach to hypertrophic scar treatment based on evidence of bacterial overload[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2026, 46(5): 585-593.

图/表 8

图1 正常皮肤和增生性瘢痕革兰染色结果Note： A. Representative images of Gram staining for normal skin and hypertrophic scar tissues are presented, illustrating different bacterial distribution patterns. B. Bacterial abundance was quantified by calculating the percentage of positively stained areas, with statistical analysis results displayed.

Fig 1 Gram staining results of normal skin and hypertrophic scar tissues

图2 不同浓度细菌干预后成纤维细胞增殖水平的变化（Edu检测）Note： A. Celluar immunofluorescence results demonstrated fibroblast proliferation after Edu detection under different concentrations of bacterial treatment. B. The ratio of Edu-positive cells to total cells was used to assess cell proliferation and the statistical results are displayed accordingly.

Fig 2 Changes in fibroblast proliferation after treatment with different bacterial concentrations detected by Edu assay

图3 不同浓度细菌干预后成纤维细胞IL-6、IL-8、TNF-α及MCP-1的分泌水平

Fig 3 Secretion levels of IL-6, IL-8, TNF-α, and MCP-1 in fibroblasts after treatment with different concentrations of bacteria

图4 不同浓度细菌处理对成纤维细胞VEGF、TGF-β1、Ⅰ型胶原和Ⅲ型胶原蛋白表达的影响Note： A. Western blotting was performed to analyze the expression of VEGF, TGF-β1, type Ⅲ collagen, and type Ⅰ collagen in fibroblasts after treatment by different concentrations of bacteria. B. The expression levels of VEGF, TGF-β1, type Ⅲ collagen, and type Ⅰ collagen were normalized to GAPDH.

Fig 4 Effects of different bacterial concentrations on the protein expression of VEGF, TGF-β1, type Ⅰ collagen, and type Ⅲ collagen in fibroblasts

图5 不同细菌浓度作用于成纤维细胞后α-SMA表达的变化Note： A. Representative immunofluorescent staining images of α-SMA in fibroblasts after treatment with different bacterial concentrations. B. The ratio of α-SMA-positive cells to total cells was used to assess α-SMA expression and the statistical results are displayed accordingly.

Fig 5 Changes in α-SMA expression in fibroblasts after exposure to different bacterial concentrations

图6 兔耳增生性瘢痕经左氧氟沙星注射后不同时相点的瘢痕外观与温哥华瘢痕量表评分Note： A. Macroscopic observations of rabbit ear hypertrophic scars at different time points after intralesional injection of levofloxacin. B. Vancouver Scar Scale (VSS) scores were compared at the corresponding time points, and the statistical results are shown.

Fig 6 Scar appearance and Vancouver Scar Scale scores at different time points after levofloxacin injection in rabbit ear hypertrophic scars

图7 左氧氟沙星注射后第60日兔耳增生性瘢痕的组织学染色Note： Representative HE staining, Masson staining, and Gram staining images of rabbit ear hypertrophic scars. The red arrows represent typical inflammatory cells, collagen fibers, and bacteria, respectively.

Fig 7 Histological staining of rabbit ear hypertrophic scars at day 60 after levofloxacin injection

图8 兔耳增生性瘢痕内注射左氧氟沙星后炎症因子IL-6、IL-8、TNF-α、MCP-1的表达情况Note： A. Immunohistochemical staining of IL-6, IL-8, TNF-α, and MCP-1 in rabbit ear hypertrophic scar tissues before and after injection of levofloxacin. E. Quantitative analysis of the relative expression levels of inflammatory cytokines.

Fig 8 Expression of inflammatory cytokines IL-6, IL-8, TNF-α, and MCP-1 in rabbit ear hypertrophic scar tissues after intralesional injection of levofloxacin

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基于“细菌超载”证据的增生性瘢痕治疗新方式探索

Novel approach to hypertrophic scar treatment based on evidence of bacterial overload

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