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目的 观察红景天甙 (salidroside,Sal)对缺氧状态下兔肺动脉平滑肌细胞 (PASMC)增殖、DNA合成和细胞周期的影响 ,并初步探讨其作用机制 .方法 应用离体培养的PASMC,采用四唑盐 (MTT)比色法实验、3H-胸腺嘧啶核苷(3H- Td R)参入和流式细胞仪技术 ,观察 PASMC的增殖、DNA合成和细胞周期的变化 .结果 缺氧 2 4h可直接刺激PASMC,使其3H- Td R的参入量显著增加 (P<0 .0 1 ) ,MTT吸光度 (A)增加 (P<0 .0 5 ) ;钙通道阻滞剂 verapamil可抑制缺氧的这种刺激作用 ;流式细胞仪分析显示缺氧 2 4h PASMC的 G2 /M期细胞比例与常氧对照组相比明显增加 (P<0 .0 1 ) ,G0 /G1 期细胞比例明显减少 ;在缺氧的 PASMC培养液中加入 Sal(0 .2～ 3.2 μmol· L- 1 ) ,PASMC的 3H- Td R参入量与 MTT的 A值与单纯缺氧组相比显著下降 ,Sal(6 .4μmol· L- 1 )可增加 G0 /G1 期细胞比例 ,减少 G2 /M期细胞比例 (P<0 .0 5 ) ;缺氧状态下 ,Sal与 verapamil合用并不增加对PASMC的抑制效应 .结论 缺氧可直接刺激 PASMC的增殖及 DNA合成 ,其作用机制可能与 PASMC内 Ca2 +浓度增加有关 ,Sal可抑制缺氧对 PASMC的增殖及 DNA合成的促进作用
Objective To investigate the effects of salidroside (Sal) on the proliferation, DNA synthesis and cell cycle of rabbit pulmonary arterial smooth muscle cells (PASMC) under hypoxia and to explore its possible mechanism.Methods PASMC cultured in vitro was treated with tetrakis (MTT) colorimetric assay, 3H-thymidine incorporation and flow cytometry were used to observe the proliferation, DNA synthesis and cell cycle changes of PASMC.Results Hypoxia 24 h could be directly Stimulation of PASMC resulted in a significant increase in the 3H-TdR incorporation (P <0.01) and MTT absorbance (A) (P <0.05). The calcium channel blocker verapamil inhibited hypoxia The results of flow cytometry showed that the proportion of cells in G2 / M phase of hypoxic 24 h PASMC increased significantly compared with normoxia control group (P <0.01), and the proportion of cells in G0 / G1 phase decreased significantly Sal (0.2-2.2 μmol · L-1) was added into hypoxic PASMC culture medium. The 3H-Td R incorporation and the MTT value of PASMC decreased significantly compared with those of hypoxia alone group. Sal (6.4 μmol · L-1) increased the proportion of cells in G0 / G1 phase and decreased the percentage of cells in G2 / M phase (P <0.05). In hypoxia, Sal and verapamil And did not increase the inhibitory effect on PASMC.Conclusion Hypoxia can directly stimulate the proliferation and DNA synthesis of PASMC, and its mechanism may be related to the increase of Ca2 + concentration in PASMC, and Sal can inhibit the promotion of PASMC proliferation and DNA synthesis by hypoxia effect