摘要
目的:观察盐酸氟桂嗪对同型半胱氨酸诱导的正常人脐静脉内皮细胞株(ECV-304)胞质内钙离子浓度和细胞活性的影响。方法:实验于2005-02/08在西京医院神经内科实验室完成。①分组:将ECV-304分为3组,正常对照组仅予空白RPMI-1640培养基培养;同型半胱氨酸组分别加入终浓度为100,200,500,1000μmol/L的同型半胱氨酸培养;盐酸氟桂嗪组加入终浓度为10μmol/L盐酸氟桂嗪及终浓度分别为100,200,500,1000μmol/L的同型半胱氨酸共培养。②细胞活性检测:用四唑盐法检测,各组细胞加入不同浓度药物培养24h后,测定各孔吸光度值A。③细胞质中[Ca2+]i的测定:各组细胞加入不同浓度药物培养20min后,利用Ca2+指示剂Flu-3AM作为细胞内钙离子的荧光探针负载培养的细胞,共聚焦技术检测细胞内荧光强度的变化。结果:①同型半胱氨酸浓度为200,500,1000μmol/L时各组的反映细胞活性A值低于对照组(0.156±0.009,0.148±0.010,0.134±0.007,0.177±0.011,P<0.01),也低于含同浓度同型半胱氨酸的盐酸氟桂嗪组(0.168±0.011,0.160±0.009,0.148±0.007,P<0.05,0.01)。且随着同型半胱氨酸浓度的增加,A值逐渐降低。②在同型半胱氨酸组中,随同型半胱氨酸浓度增加反映细胞质中[Ca2+]I的平均荧光强度也逐渐增强。同型半胱氨酸各浓度组平均荧光强度均高于对照组(P<0.01);也高于含相同浓度同型半胱氨酸的盐酸氟桂嗪组(P<0.01)。结论:①同型半胱氨酸对血管内皮细胞有损伤作用,其作用与胞内钙离子浓度升高密切相关。②盐酸氟桂嗪能减轻同型半胱氨酸所诱导的细胞损伤,可能与其直接抑制由同型半胱氨酸所诱导的Ca2+的内流,减轻细胞质钙超载有关。
AIM： To observe the effects of flunarizine hydrochloride （FZ） on intracytoplasm calcium ionic concentration and cell activity of endothelial cell strain （ECV-304） induced by homocysteine （Hcy）. METHODS： The experiment was conducted at Neurological Laboratory of Xijing Hospital between February and August 2005.①Group： ECV-304 were divided into 3 groups： Control group （ECV-304 cells were cultured only in RPMI-1640 culture medium）, Hey group （ECV-304 cells were cultured in Hey with final concentrations of 100,200,500 and 1 000 μmol/L） and treatment group （ECV-304 cells were co-cuhured in FZ with final concentrations of 10 μmol/L and Hey of different concentrations of 100, 200,500 and 1 000 μmol/L）. ②Detection of cell activity： Tetrazolium was used to detect absorbance. （A） of each hole in three groups after 24-hour culture with drugs of different concentrations. ③Detection of intraeytoplasm calcium ions： After ECV-304 were cultured for 20 minutes with different concentrations of drugs and loaded With Ca^2＋ indicator Flu-3AM as fluorescent probe, the intracellular fluorescent intensity （FI） was measured with eonfocal microscopy. RESULTS： ①When Hey was concentrated at 200,500 and 1 000 μmol/L, A value reflecting cell activity was lower in Hcy group than in the control group （0.156±0.009, 0.148±0.010, 0.134±0.007, 0.177±0.011, P 〈 0.01） and treatment group cotitaining same concentration of Hcy （0.168±0.011, 0.160±0.009, 0.148±0.007, P 〈 0.05,0.01）. And A value decreased gradually as Hey concentration increased. ②In Hcy group, the average FI of calcium ions in cytoplasm advanced gradually with the increased concentration of Hey, and was higher than that of control group and treatment group containing same concentration of Hcy （P 〈 0.01）.
CONCLUSION： ①Hey can damage the endothelial cells of blood vessels, with the mechanism closely related with the enhancement of intracellular calcium ionic concentration. ②FZ has the protective
出处
《中国临床康复》
CSCD
北大核心
2006年第17期92-94,共3页
Chinese Journal of Clinical Rehabilitation
