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目的 :研究红景天甙 (Salidroside ,Sal)对低氧 ( 3 % )条件下兔肺动脉平滑肌细胞 (PASMC)增殖、DNA合成、细胞内钙离子浓度的影响。方法 :应用细胞培养、四唑盐比色试验 (MTT)、[3H]-胸腺嘧啶核苷 ( [3H]-TdR)掺入试验、Fluo- 3和激光扫描共聚焦显微术测定细胞内Ca2 +浓度等方法。结果 :低氧 2 4h可直接刺激PASMC ,使MTT的A值增加 62 % (P <0 0 5 ) ,[3H]-TdR掺入量增加 13 8% (P <0 0 1)。在低氧的PASMC培养液中加入Sal( 3 2× 10 -5mol/L)可抑制低氧的这种促增殖作用 ,与低氧组相比MTT的A值下降 2 9% (P <0 0 5 ) ,[3H]-TdR掺入量下降3 7%。在低氧的PASMC培养液中加入钙通道阻滞剂verapamil也可抑制低氧的促增殖作用 ,与低氧组相比较 ,MTT的A值下降 2 3 % (P <0 0 5 ) ,[3H]-TdR掺入量下降 5 1% (P <0 0 1)。PASMC的低氧培养可使细胞内Ca2 +浓度明显增加 (P <0 0 5 ) ,Sal可抑制低氧所致的细胞内Ca2 +浓度的上升。结论 :红景天甙可抑制低氧促进PASMC增殖、DNA合成的作用 ,其机制可能与抑制低氧时细胞内Ca2 +浓度增加有关。
Objective: To study the effects of salidroside (Saldroside, Sal) on proliferation, DNA synthesis, and intracellular calcium concentration in rabbit pulmonary artery smooth muscle cells (PASMC) under hypoxic (3%) conditions. METHODS: Cell culture, tetrazolium salt colorimetric assay (MTT), [3H]-thymidine ([3H]-TdR) incorporation assay, Fluo-3 and laser scanning confocal microscopy were used to determine intracellular Ca2 + concentration and other methods. RESULTS: PASMCs were stimulated directly with hypoxia for 24 hours, which increased MTT A value by 62% (P <0 05) and increased [3H]-TdR incorporation by 13% (P <0 01). Adding Sal (3 2 × 10 -5 mol/L) to hypoxic PASMC cultures inhibited this pro-proliferative effect of hypoxia. Compared with the hypoxic group, the A value of MTT decreased by 29% (P < 0 0 5) The [3H]-TdR incorporation decreased by 37%. The addition of the calcium channel blocker verapamil in the hypoxic PASMC medium also inhibited the hypoxic proliferative effect. Compared with the hypoxic group, the MTT A value decreased by 2 3 % (P <0 05). ]-TdR incorporation decreased by 5 1% (P < 0 01). Hypoxic culture of PASMC significantly increased intracellular Ca2+ concentration (P < 0.05), and Sal inhibited the increase of intracellular Ca2 + concentration induced by hypoxia. Conclusion : Salidroside can inhibit the hypoxia-induced proliferation and DNA synthesis of PASMC, and its mechanism may be related to the inhibition of intracellular Ca2+ concentration in hypoxia.