目的 探讨祛风汤对系统性硬化症（SSc）皮肤纤维化模型小鼠的干预作用及分子机制。方法 采用sc博来霉素建立SSc皮肤纤维化小鼠模型，随机分为对照组、模型组、尼达尼布（NTD，39 mg·kg^-1）组及祛风汤低、中、高剂量（3.9、7.8、15.6 g·kg^-1）组，连续ig给药28 d。通过苏木素-伊红（HE）染色、Masson染色及飞秒无标记成像观察皮肤组织病理变化；免疫组化检测α-平滑肌肌动蛋白（α-SMA）和转化生长因子-β（TGF-β）表达； ELISA法检测羟脯氨酸（HYP）及炎症因子肿瘤坏死因子（TNF）-α、白细胞介素（IL）-6、IL-1β含量；转录组学筛选差异表达基因及富集通路； Western blotting检测单磷酸腺苷活化蛋白激酶（AMPK）/过氧化物酶体增殖物激活受体-γ（PPAR-γ）/脂联素（Adiponectin）通路蛋白表达。结果 与模型组比较，祛风汤各剂量组小鼠真皮厚度显著变薄（P＜0.01、0.001），胶原纤维沉积减少、排列趋于疏松，皮下脂肪层有所恢复，炎症细胞浸润减轻； α-SMA和TGF-β蛋白表达下调（P＜0.05）；飞秒无标记成像显示胶原纤维表达量呈剂量相关性下降；祛风汤高剂量组皮肤组织中HYP含量及TNF-α、IL-6、IL-1β水平显著降低（P＜0.05、0.01、0.001）；转录组学分析显示，差异表达基因显著富集于PPAR-γ相关信号通路； Western blotting结果显示，与对照组比较，模型组小鼠皮肤组织中p-AMPK/AMPK、PPAR-γ及Adiponectin蛋白表达显著下调（P＜0.01、0.001），而祛风汤中、高剂量组上述蛋白表达显著上调（P＜0.05、0.01、0.001）。结论 祛风汤可改善博来霉素诱导的皮肤纤维化，其机制与AMPK/PPAR-γ/Adiponectin信号通路激活相关，可能涉及该通路对TGF-β介导的成纤维细胞活化及胶原异常沉积的调控。

Objective To investigate the ameliorative effect of Qufeng Decoction on bleomycin-induced dermal fibrosis mouse model and to elucidate the underlying molecular mechanisms. Methods A mouse model of SSc skin fibrosis was established by subcutaneous injection of bleomycin (BLM). The mice were randomly divided into control, model, nintedanib (NTD, 39 mg·kg^-1), and Qufeng Decoction low-, medium-, and high-dose (3.9, 7.8, and 15.6 g·kg^-1) groups, and were ig administered for 28 consecutive days. Skin histopathological changes were evaluated by hematoxylin-Eosin (HE) staining, Masson staining, and femtosecond label-free imaging. Immunohistochemistry was used to detect the expression of α-smooth muscle actin (α-SMA) and transforming growth factor- β (TGF-β). Hydroxyproline (HYP) and inflammatory cytokines (TNF-α, IL-6, and IL-1β) in skin tissue were measured by ELISA. Transcriptomic analysis was performed to screen differentially expressed genes (DEGs) and enriched pathways. Western blotting was conducted to examine protein expression in the AMPK/PPAR-γ/Adiponectin pathway. Results Compared with the model group, Qufeng Decoction treatment significantly reduced dermal thickness in all dose groups (P < 0.01, 0.001), attenuated collagen fiber deposition with a more loosely arranged pattern, partially restored the subcutaneous fat layer, and alleviated inflammatory cell infiltration. The protein expression of α-SMA and TGF-β was significantly downregulated (P < 0.05). Femtosecond label-free imaging revealed a dose-dependent decrease in collagen fiber content. In the Qufeng Decoction high-dose group, skin HYP content and levels of TNF-α, IL-6, and IL-1β were significantly decreased (P < 0.05, 0.01, and 0.001). Transcriptomic analysis showed that differentially expressed genes were significantly enriched in peroxisome proliferator-activated receptor-γ (PPAR-γ)-related signaling pathways. Western blotting results indicated that the ratios of p-AMPK/AMPK, as well as PPAR-γ and Adiponectin protein expression, were significantly downregulated in the model group (P < 0.01 and 0.001), whereas Qufeng Decoction at medium and high doses significantly reversed these changes (P < 0.05, 0.01, and 0.001). Conclusion Qufeng Decoction ameliorates BLM-induced skin fibrosis in mice, and the mechanism is associated with activation of the AMPK/PPAR-γ/Adiponectin signaling pathway, potentially involving the modulation of TGF-β-mediated fibroblast activation and abnormal collagen deposition.

R965

解放军总医院第六医学中心创新培育基金资助项目（CXPY202406）；中央级公益性科研院所基本业务费专项资金资助（ZXKT22045，ZXKT25006）；中国中医科学院科技创新工程项目（CI2026A04618）；中国中医科学院优秀青年科技人才培养专项（ZZ16-YQ-027）