目的 考察三七总皂苷（PNS）的体内外抗肺纤维化作用，并探讨其作用机制。方法 将60只Wistar大鼠随机分为假手术组、模型组、吡非尼酮（阳性药，50 mg·kg^-1）组和PNS低、中、高剂量（50、100、200 mg·kg^-1）组，采用气管内注入博来霉素（5 mg·kg^-1）建立肺纤维化大鼠模型，假手术组注入生理盐水；造模24 h后给药，持续28 d；检测大鼠肺脏系数，利用BUXCO系统检测大鼠气道阻力与肺顺应性变化，HE染色后观察大鼠肺组织病理结构损伤，免疫荧光法检测大鼠肺组织E-钙黏蛋白（E-cad）、N-钙黏蛋白（N-cad）表达，免疫组化法检测大鼠肺组织蛋白酶激活受体-1（PAR-1）蛋白表达；体外培养人胚肺成纤维细胞MRC-5，设对照组、模型组（凝血酶2 U·mL^-1建立体外肺纤维化模型）和PNS 5、10、20 μg·mL^-1组，体外划痕实验检测细胞迁移率，实时荧光定量PCR（qRT-PCR）、Western blotting实验检测α-平滑肌肌动蛋白（α-SMA）、波形蛋白（Vim）和PAR-1基因和蛋白表达水平；随后使用PAR-1 si RNA抑制PAR-1表达后，进一步采用qRT-PCR和Western blotting检测α-SMA与Vim基因与蛋白表达。结果 体内实验中，与模型组相比，PNS各剂量组肺脏系数显著降低（P＜0.001）、肺顺应性显著升高（P＜0.05、0.01、0.001），100、200 mg·kg^-1 PNS组大鼠气道阻力显著降低（P＜0.05、0.01），同时PNS能够改善大鼠肺组织的病理结构损伤，在下调N-cad的蛋白表达同时上调E-cad的蛋白表达，且100、200 mg·kg^-1 PNS组PAR-1蛋白表达显著下调（P＜0.01、0.001）。体外实验中，与模型组相比，10、20 μg·mL^-1 PNS组细胞的迁移率显著降低（P＜0.01、0.001），20 μg·mL^-1 PNS组α-SMA与Vim mRNA水平下调（P＜0.05、0.01），20 μg·mL^-1 PNS组α-SMA蛋白表达水平显著下调（P＜0.05） ，10、20 μg·mL^-1 PNS组Vim蛋白表达水平显著下调（P＜0.05、0.01），PAR-1 siRNA组α-SMA mRNA水平和α-SMA、Vim蛋白表达水平显著降低（P＜0.05、0.01、0.001）。结论 PNS具有抗肺纤维化的作用，其机制可能与调控PAR-1的异常有关。

Objective To investigate the in vivo and in vitro anti pulmonary fibrosis effects of Panax notoginseng saponins (PNS) and preliminarily explore its pharmacological mechanism. Methods Totally 60 Wistar rats were randomly divided into sham-surgery group, model group, pirfenidone (positive drug, 50 mg·kg^?1) group, and PNS (50, 100, and 200 mg·kg^?1) group. A pulmonary fibrosis rat model was established by tracheal injection of bleomycin (5 mg·kg^?1), physiological saline were injected into rats in shamsurgery group. After 24 hours, the treatment group was given corresponding drugs once a day for 28 d. The lung coefficient of rats were detected, the BUXCO system was used to detect changes in airway resistance and lung compliance, pathological structural damage of rat lung tissue was observe after HE staining, the expression of E-cadherin (E-cad) and N-cadherin (N-cad) in rat lung tissue were detected using immunofluorescence method, and the expression of tissue protease activated receptor-1 (PAR-1) protein in rat lung tissue were detected using immunohistochemistry method. Meanwhile, human embryonic lung fibroblasts MRC-5 were cultured in vitro, with control group, model group (thrombin 2 U·mL^?1 to establish a pulmonary fibrosis model) and PNS 5, 10, and 20 μg·mL^?1 group were set up, in vitro scratch assay to detect cell migration rate, real-time fluorescence quantitative PCR (qRTPCR) and Western blotting assay were used to detect the expression levels of α -smooth muscle actin (α -SMA), Vim, and PAR-1 genes and proteins. Subsequently, after inhibiting PAR-1 expression using PAR-1 si-RNA, qRT-PCR and Western blotting were further used for detection α- SMA and Vim gene and protein expression. Results In vivo experiments, compared with model group, the lung index of the 50, 100, and 200 mL·kg^?1 PNS groups decreased (P < 0.001), the lung compliance of the 50, 100 and 200 mg·kg^?1 PNS groups significantly increased (P < 0.05, 0.01, 0.001), while the airway resistance of rats in the 100 and 200 mg·kg^?1 PNS groups significantly decreased (P < 0.05, 0.01). PNS improved the pathological structural damage of rat lung tissue, downregulated the protein expression of N-cad, upregulated the protein expression of E-cad, and the expression of PAR-1 protein in the 100 and 200 mg·kg^?1 PNS groups were significantly downregulated (P < 0.05, 0.01). In vitro experiments, compared with the model group, the migration rate of 10 and 20 μg·mL^?1 PNS group cells was significantly reduced (P < 0.01, 0.001), α-SMA and Vim mRNA levels in 20 μg·mL^?1 PNS group were downregulated (P < 0.05, 0.01), the α-SMA protein expression level in 20 μg·mL^?1 PNS group was significantly downregulated (P < 0.05), while the Vim protein expression level in the 10 and 20 μg·mL-1 PNS group was significantly downregulated (P < 0.05, 0.01), the expression levels of α-SMA mRNA and SMA and Vim proteins were significantly reduced (P < 0.05, 0.01, 0.001). Conclusion PNS have anti-pulmonary fibrosis effects, and its mechanism may be related to the regulation of PAR-1.

R285.5

国家自然科学基金青年项目（82204740）；中国中医科学院中药研究所中药药理创新团队项目（CI2021B015）；中国中医科学院优秀青年科技人才培养专项（ZZ16-YQ-027）