低分子肝素抑制bFGF诱导犬动脉平滑肌细胞增殖的体外实验研究

The Inhibitory Effect of Low Molecular Weight Heparin on bFGF-induced Proliferation of Canine Arterial Smooth Muscle Cell: An Experimental Study in Vitro

目的 明确低分子肝素 (LMWH)对碱性成纤维细胞生长因子 (bFGF)诱导培养的犬动脉平滑肌细胞增殖的抑制作用。进一步探讨LMWH作用的最佳有效浓度。材料与方法 在 10ng/mlbFGF刺激下 ,一定数量的第 5代犬动脉平滑肌细胞 (5× 10 3 /孔 )含 0、10、5 0、10 0、2 0 0、40 0、80 0、10 0 0、2 0 0 0antiXaIU/ml各浓度组LMWH的DMEM +10 %FBS细胞培养液中孵育 72h ,MTT比色计量法测出各组吸收光密度 (OD)值 ,计算抑制率 ,选择最佳抑制浓度。结果 LMWH对bFGF诱导平滑肌细胞增殖的抑制与其浓度呈线性正相关。 10 0 0antiXaIU/ml浓度的抑制率达 5 0 %以上 ,为显著有效。结论 进一步验证了LMWH对PTA术后多种细胞因子 (尤其是bFGF)局部刺激下的平滑肌细胞增殖的抑制作用。

Objective To identify the inhibitory effect of low molecular weight heparin (LMWH) on the proliferation of canine arterial smooth muscle cells induced by bFGF in vitro, and to assess the optimal effective concentration of LMWH.Materials and Methods A certain amount (5×10 3/well) of vascular smooth muscle cells (VSMCs) were cultured in DMEM+10% FBS medium containing 10ng/ml bFGF. The cell culture medium with different LMWH concentration, i.e. 0, 10, 50, 100, 200, 400, 800, 1000 and 2000 anti-XaIU/ml, was prepared separately. After 72 hours incubation, the OD values of different groups were calculated by using MTT method. The inhibitory rate was calculated and the optimal concentration of LMWH was determined.Results A linear positive correlation existed between the inhibitory effect of LMWH and its concentration. The inhibitory rate of 1000 anti-XaIU was over 50%.Conclusion This study further validates the inhibitory effect of LMWH on the VSMCs' proliferation induced by multiple cell factors, especially bFGF, after PTA, and provides reliable experimental evidence for the clinical application of LMWH to prevent vascular restenosis after PTA.