目的 基于网络药理学和动物实验验证探究芦根Phragmites communis改善肺纤维化的活性成分、关键靶点及作用机制，并比较鲜芦根与干芦根的药效差异。方法 利用中药系统药理学数据库与分析平台（TCMSP）、Herb数据库获取芦根活性成分及对应靶点，借助Genecards、TTD、OMIM数据库筛选肺纤维化疾病靶点；二者交集靶点用于构建网络；筛选核心靶点并开展基因本体（GO）和京都基因与基因组百科全书（KEGG）分析。单次气管内滴注博来霉素（2.3 mg·kg^-1）构建肺纤维化模型，将小鼠随机分为对照组，模型组，鲜芦根水煎液（FPCAE）低、高剂量组（5.2、10.4 g·kg^-1），干芦根水煎液（DPCAE）低、高剂量（2.6、5.2 g·kg^-1）组和吡非尼酮（阳性药，234 mg·kg^-1）组。造模次日起连续ig给药21 d。比较各组小鼠的肺脏质量和肺脏指数；通过苏木素-伊红（HE）、Masson染色及Szapiel、Ashcroft评分对小鼠肺组织病理形态进行评估；碱水解法检测小鼠血清羟脯氨酸（HYP）水平；实时荧光定量PCR（qRT-PCR）法检测小鼠肺组织中白细胞介素-6（Il-6）、白细胞介素-1β（Il-1β）、肿瘤坏死因子（Tnf-α）、E-钙黏蛋白（Cdh1）、α-平滑肌肌动蛋白（Acta2）、Ⅰ型胶原（Col1a1）和Ⅲ型胶原（Col3a1）的mRNA表达； Western blotting法检测小鼠肺组织中E-钙黏蛋白（E-cadherin）、α-平滑肌肌动蛋白（α-SMA）和磷酸化磷脂酰肌醇3-激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白（PI3K/Akt/mTOR）信号通路相关蛋白的表达。结果 网络药理学中筛选得到芦根与肺纤维化疾病的交集靶点有499个，核心靶点主要涵盖磷脂酰肌醇-3-激酶调节亚基1（PIK3R1）、蛋白激酶B（Akt1）、磷脂酰肌醇3-激酶（PIK3CA）、信号转导与转录激活因子3（STAT3）等，KEGG富集结果显示PI3K-Akt、丝裂原活化蛋白激酶（MAPK）、晚期糖基化终产物-晚期糖基化终产物受体（AGE-RAGE）等信号通路可能参与芦根干预肺纤维化过程。动物实验结果显示，与模型组比较，芦根鲜品与干品均可降低小鼠的肺脏质量、肺脏指数及血清HYP含量，减轻肺组织炎症细胞浸润、肺泡结构破坏及胶原纤维沉积，降低Szapiel、Ashcroft评分；下调肺组织Il-6、Il-1β、Tnf-α、Acta2、Col1a1和Col3a1 mRNA表达，上调Cdh1 mRNA表达；升高E-cadherin蛋白表达，减少α-SMA及p-PI3K、p-Akt、p-mTOR蛋白表达（P＜0.05、0.01、0.001）。FPCAE较DPCAE在降低肺脏指数、炎症因子表达、胶原沉积及调控PI3K/Akt/mTOR通路方面整体呈现更明显改善趋势。结论 芦根鲜品与干品均可有效改善博来霉素诱导的小鼠肺纤维化，其作用机制可能与抑制PI3K/Akt/mTOR信号通路活化，进而减轻炎症反应、上皮间质转化进程及细胞外基质沉积有关，且鲜品疗效更佳。

Objective To investigate the active constituents, key targets, and underlying mechanisms of Phragmitis Rhizoma in the treatment of pulmonary fibrosis based on network pharmacology and experimental validation, and to compare the pharmacological effects of fresh and dried Phragmitis Rhizoma. Methods Active constituents and corresponding targets of Phragmitis Rhizoma were collected from the TCMSP and HERB databases. Pulmonary fibrosis-related targets were retrieved from the GeneCards, TTD, and OMIM databases. The overlapping targets between drug-related and disease-related targets were identified and used to construct a protein-protein interaction (PPI) network. Core targets were screened, followed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. Mice were randomly assigned to a normal control group, model group, fresh Phragmitis Rhizoma (FPCAE) low-dose group (5.2 g·kg^-1), FPCAE high-dose group (10.4 g·kg^-1), dried Phragmitis Rhizoma (DPCAE) low-dose group (2.6 g·kg^-1), DPCAE high-dose group (5.2 g·kg^-1), and pirfenidone group (234 mg·kg^-1). Except for the normal control group, pulmonary fibrosis was induced by a single intratracheal instillation of bleomycin (2.3 mg·kg^-1). Drug administration was initiated on the day following modeling and continued for 21 consecutive days. Lung weight and lung coefficient were determined. Histopathological alterations were evaluated by hematoxylin-eosin (HE) staining, Masson staining, and Szapiel and Ashcroft scoring. Serum hydroxyproline (HYP) levels were measured using the alkaline hydrolysis method. The mRNA expression levels of interleukin-6 (Il-6), interleukin-1β (Il-1β), tumor necrosis factor-α (Tnf-α), E-cadherin (Cdh1), α-smooth muscle actin (Acta2), collagen type I alpha 1 (Col1a1), and collagen type III alpha 1 (Col3a1) were detected by RT-qPCR. Protein expression levels of Ecadherin, α-smooth muscle actin (α-SMA), and PI3K/Akt/mTOR signaling pathway-related proteins were determined by Western blotting. Results Network pharmacology analysis identified 499 overlapping targets between Phragmitis Rhizoma and pulmonary fibrosis. Key targets mainly included PIK3R1, Akt1, PIK3CA, and STAT3. KEGG enrichment analysis suggested that the PI3K-Akt, MAPK, and AGE-RAGE signaling pathways may be involved in the anti-pulmonary fibrosis effects of Phragmitis Rhizoma. Compared with the model group, both fresh and dried Phragmitis Rhizoma reduced lung weight, lung coefficient, and serum HYP levels, alleviated inflammatory cell infiltration, alveolar structural destruction, and collagen deposition in lung tissues, and decreased Szapiel and Ashcroft scores. In addition, treatment downregulated the mRNA expression levels of Il-6, Il-1β, Tnf-α, Acta2, Col1a1, and Col3a1, while upregulating Cdh1 expression. Moreover, E-cadherin expression was increased, whereas α-SMA, p-PI3K, p-Akt, and p-mTOR protein expression levels were decreased (P < 0.05, 0.01, 0.001). Compared to DPCAE, FPCAE showed a more pronounced improvement trend in reducing lung index, inflammatory factor expression, collagen deposition, and regulating the PI3K/Akt/mTOR pathway. Conclusion Both fresh and dried Phragmitis Rhizoma effectively ameliorated bleomycin-induced pulmonary fibrosis in mice. The underlying mechanism may be associated with inhibition of PI3K/Akt/mTOR signaling pathway activation, thereby attenuating inflammatory responses, suppressing epithelial-mesenchymal transition, and reducing extracellular matrix deposition. Fresh Phragmitis Rhizoma exhibited superior therapeutic efficacy compared with the dried form.

R965

深圳市医疗卫生三名工程“深圳市宝安区中医院-广州中医药大学刘中秋教授中药制剂开发及转化药学研究团队”项目（SZZYSM202206005）；浙江省中医药科技计划项目（2025ZR014）