OBJECTIVE To investigate the effect of asiaticoside (AS) on endothelial-to-mesenchymal transition (EndoMT) in hypoxia pulmonary hypertension (HPH). METHODS Male Sprague-Dawley (SD) rats were divided into normoxia control group, hypoxia model group, and AS 25 and 50 mg · kg-1 group. Hypoxia model group and AS group were subjected to intermittent hypoxia exposure. Control group and model group received 1-1.5 mL saline daily, and AS groups were ig administrated with AS 25 and 50 mg·kg-1 for 4 weeks. Human pulmonary artery endothelial cells (HPAECs) were divided into normoxia control group and hypoxia AS groups. Hypoxia groups were cultured with AS 0, 25, 50, 100 and 200 mg·L-1 for 72 h under hypoxic (5%O2, 5%CO2) conditions. Anti-proliferation effect of AS was investigated by CCK-8 assay. Then, HPAECs were divided into normoxia control group, normoxia AS 100 mg · L-1 group, hypoxia model group, and hypoxia AS 100 mg · L-1 group. After five days of culture, migration ability of cells was detected by Transwell test. Expression of CD31 andα-SMA was detected by immunofluorescence and Western blotting in both in vivo and in vitro experiments. RESULTS In both in vivo and in vitro experiments, compared with normoxia control group, expression of CD31 was reduced (P<0.01) andα-smooth muscle actin (α-SMA) was increased (P<0.01) in hypoxia model group in both immunofluorescent analysis and Western blotting. Compared with hypoxia model group, expression of CD31 was increased andα-SMA was decreased (P<0.05, P<0.01) in AS treatment groups. Compared with normoxia control group, proliferation and migration ability of HPAEC were elevated in hypoxia model group (P<0.05). Compared with hypoxia group, AS 100 mg · L-1 depressed proliferation and migration of HPAEC under hypoxia exposure up to 72 h (P<0.05). CONCLUSION EndoMT might be involved in HPH and could be partly inhibited by AS.%目的 研究内皮-间充质转化在低氧肺动脉高压的作用及积雪草苷(AS)对内皮-间充质转化的影响.方法 ①体内实验:雄性SD大鼠分为常氧对照组、低氧模型组、AS 25和50 mg·kg-1给药组.间歇低氧法(每天低氧8 h,每周6 d,持续4周)制备HPH大鼠模型,AS组用AS 25或50 mg·kg-1·d-1治疗4周.②体外实验:将人肺动脉内皮细胞(HPAEC)分为常氧组和低氧模型组,低氧模型组在低氧条件下(5%O2,5%CO2)培养,用AS 0,25,50,100或200 mg·L-1作用72 h,CCK-8法检测HPAEC存活情况;再将HPAEC分为常氧对照组、常氧AS 100 mg·kg-1组、低氧模型组和低氧AS 100 mg·kg-1组,细胞培养5 d后用Tran?swell法检测HPAEC迁移能力,免疫荧光及Western蛋白印迹检测CD31和α平滑肌肌动蛋白(α-SMA)蛋白的表达水平.结果 在体内和体外实验中,免疫荧光和Western蛋白印迹结果均显示,与常氧对照组相比,低氧模型组的CD31表达下降(P<0.01),α-SMA表达升高(P<0.01);与低氧模型组相比,AS给药组CD31表达升高,α-SMA表达减少(P<0.05,P<0.01).HPAEC的增殖及迁移能力结果显示,与常氧对照组相比,低氧模型组HPAEC的增殖及迁移能力升高;AS 100 mg·L-1能抑制低氧培养72 h后的HPAEC的增殖和迁移能力(P<0.05).结论 HPH可能与内皮-间充质转化有关,AS对此有一定的抑制作用.
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