水蛭素对体外培养的兔血管内皮细胞及平滑肌细胞生长的影响

摘要

目的 探讨水蛭素对体外培养的兔血管内皮细胞及平滑肌细胞生长的影响及剂量依赖关系.方法 用体外培养兔血管内皮细胞及平滑肌细胞第3 ～5代,加入96孔细胞培养板,同时加入不同浓度的水蛭素0.78 、1.56、3.13 、6.25 、12.5 、25 mg/L,每6孔为一个浓度组,1640培养液为空白对照组,共7组,培养48 h后用噻唑蓝比色试验(MTT法)测定各孔贴壁细胞的吸光值(OD).结果 与空白对照组相比,低、中、高浓度水蛭素作用于体外培养的兔血管内皮细胞后,细胞形态无明显改变,贴壁良好,生长无明显抑制作用(P> 0.05);而水蛭素作用于兔血管平滑肌细胞后,与空白对照组相比,低浓度水蛭素对平滑肌细胞形态无明显改变,中、高浓度水蛭素作用于平滑肌细胞后,细胞密度有所减小、排列松散,OD值下降,呈现抑制平滑肌细胞的生长(P<0.05).结论 不同浓度水蛭素对体外培养兔血管内皮细胞生长无显著抑制作用;低浓度水蛭素对体外培养兔血管平滑肌细胞生长无明显抑制作用,中、高浓度水蛭素可抑制兔血管平滑肌细胞的生长.%Objective To investigate the effects of different doses of hirudin on the growth of rahbit vascular endothelial cells( EC ) and smooth musele cells ( SMC ) . Methods The 3～ 5 generation of rabbit vascular endothelial cells( EC ) and smooth musele cells ( SMC ) were respectively incubated in a 96-well culture plate. then co-incubated with different concentration of hirudin( 0. 78 , 1. 56. 3. 13. 6. 25 . 12. 5,25 mg/L ) . every dose added into 6 holes , the first 6 holes were added with RPMl-1640 as control group. After co-incubation of 48 hours the growth artivity of the 7 groups of cells were tested by MTT colorimetric method for the optical densitv difference(OD ) value. Results Compared to the RPMl-1640 control group,no apparent changes of cell morphology were observed in the ECs cultivated in low, medium and high concentration hirudin solution, good adherence, no obvious inhibition of the growth of ECs( P ＞0. 05 ); as for the vascular SMCs . low concentration of hirudin had no apparent effects on the growth of vascular SMCs , medium and high concentration of hirudin inhihited the growth of vascular SMCs.lowered the cells density and the OD value( P ＜0. 05 ), presenting an inhibitive effect on SMC growth. Conclusion hirudin of different concentrations have no apparent effects on the growth of the rabbit ECs; hirudin of low concentration has no apparent effects on the growth of the rabbit vascular SMCs,while the medium and high concentrations present an inhihition effect on the rabbit vascular SMC growth.