目的 考察三七總皂苷（PNS）的體內(nèi)外抗肺纖維化作用，并探討其作用機(jī)制。方法 將60只Wistar大鼠隨機(jī)分為假手術(shù)組、模型組、吡非尼酮（陽(yáng)性藥，50 mg·kg^-1）組和PNS低、中、高劑量（50、100、200 mg·kg^-1）組，采用氣管內(nèi)注入博來(lái)霉素（5 mg·kg^-1）建立肺纖維化大鼠模型，假手術(shù)組注入生理鹽水；造模24 h后給藥，持續(xù)28 d；檢測(cè)大鼠肺臟系數(shù)，利用BUXCO系統(tǒng)檢測(cè)大鼠氣道阻力與肺順應(yīng)性變化，HE染色后觀察大鼠肺組織病理結(jié)構(gòu)損傷，免疫熒光法檢測(cè)大鼠肺組織E-鈣黏蛋白（E-cad）、N-鈣黏蛋白（N-cad）表達(dá)，免疫組化法檢測(cè)大鼠肺組織蛋白酶激活受體-1（PAR-1）蛋白表達(dá)；體外培養(yǎng)人胚肺成纖維細(xì)胞MRC-5，設(shè)對(duì)照組、模型組（凝血酶2 U·mL^-1建立體外肺纖維化模型）和PNS 5、10、20 μg·mL^-1組，體外劃痕實(shí)驗(yàn)檢測(cè)細(xì)胞遷移率，實(shí)時(shí)熒光定量PCR（qRT-PCR）、Western blotting實(shí)驗(yàn)檢測(cè)α-平滑肌肌動(dòng)蛋白（α-SMA）、波形蛋白（Vim）和PAR-1基因和蛋白表達(dá)水平；隨后使用PAR-1 si RNA抑制PAR-1表達(dá)后，進(jìn)一步采用qRT-PCR和Western blotting檢測(cè)α-SMA與Vim基因與蛋白表達(dá)。結(jié)果 體內(nèi)實(shí)驗(yàn)中，與模型組相比，PNS各劑量組肺臟系數(shù)顯著降低（P＜0.001）、肺順應(yīng)性顯著升高（P＜0.05、0.01、0.001），100、200 mg·kg^-1 PNS組大鼠氣道阻力顯著降低（P＜0.05、0.01），同時(shí)PNS能夠改善大鼠肺組織的病理結(jié)構(gòu)損傷，在下調(diào)N-cad的蛋白表達(dá)同時(shí)上調(diào)E-cad的蛋白表達(dá)，且100、200 mg·kg^-1 PNS組PAR-1蛋白表達(dá)顯著下調(diào)（P＜0.01、0.001）。體外實(shí)驗(yàn)中，與模型組相比，10、20 μg·mL^-1 PNS組細(xì)胞的遷移率顯著降低（P＜0.01、0.001），20 μg·mL^-1 PNS組α-SMA與Vim mRNA水平下調(diào)（P＜0.05、0.01），20 μg·mL^-1 PNS組α-SMA蛋白表達(dá)水平顯著下調(diào)（P＜0.05） ，10、20 μg·mL^-1 PNS組Vim蛋白表達(dá)水平顯著下調(diào)（P＜0.05、0.01），PAR-1 siRNA組α-SMA mRNA水平和α-SMA、Vim蛋白表達(dá)水平顯著降低（P＜0.05、0.01、0.001）。結(jié)論 PNS具有抗肺纖維化的作用，其機(jī)制可能與調(diào)控PAR-1的異常有關(guān)。

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

國(guó)家自然科學(xué)基金青年項(xiàng)目（82204740）；中國(guó)中醫(yī)科學(xué)院中藥研究所中藥藥理創(chuàng)新團(tuán)隊(duì)項(xiàng)目（CI2021B015）；中國(guó)中醫(yī)科學(xué)院優(yōu)秀青年科技人才培養(yǎng)專項(xiàng)（ZZ16-YQ-027）