摘要
目的:观察盐酸氟桂利嗪对同型半胱氨酸诱导的人脐静脉内皮细胞株黏附分子表达的影响。方法:实验于2005-02/08在西京医院神经内科实验室完成。人脐静脉内皮株细胞培养,取第4～7代指数生长期细胞,接种到96孔板,24h细胞贴壁后换液,加入药物处理。随机分为正常对照组、同型半胱氨酸组及盐酸氟桂利嗪组3组,正常对照组,仅予空白RPMI-1640培养基培养24h;同型半胱氨酸组,分别加入终浓度为100μmol/L、200μmol/L、500μmol/L、1000μmol/L的同型半胱氨酸培养24h;盐酸氟桂利嗪组:加入终浓度为10μmmol/L盐酸氟桂利嗪及终浓度分别为为100μmol/L、200μmol/L、500μmol/L、1000μmol/L的同型半胱氨酸,培养24h。以上每组各种浓度共8孔。用ELISA法检测各组不同培养条件下可溶性细胞间黏附分子1和可溶性血管细胞黏附因子1的吸光度值(波长490nm)。结果:同型半胱氨酸组中随着培养液同型半胱氨酸浓度的增加可溶性细胞间黏附分子1的吸光度值从(0.143±0.013)增至(0.175±0.006),与对照组相比差异有显著性意义(P<0.01);可溶性血管黏附分子1从(0.112±0.008)增至(0.147±0.014),与对照组相比差异有显著性意义(P<0.01);10μmmol/L的盐酸氟桂利嗪+100μmol/L同型半胱氨酸组与相同浓度的同型半胱氨酸组相比吸光度值也减少,差异无显著性意义(P>0.05);10μmmol/L的盐酸氟桂利嗪+200μmol/L、500μmol/L、1000μmol/L同型半胱氨酸组的吸光度均低于相同浓度的同型半胱氨酸组,差异有显著性意义(P<0.01);表明盐酸氟桂利嗪明显抑制同型半胱氨酸所诱导的可溶性细胞间黏附分子1和可溶性血管细胞黏附因子1的表达。结论:盐酸氟桂利嗪能够减少同型半胱氨酸引起的人脐静脉内皮细胞株黏附分子的上调,可能对细胞具有保护作用。
AIM： To observe the effect of flunarizine hydrochloride on the expressions of cell adhesion molecules in human umbilical vein endothelial cell line （ECV-304 cells） induced by homocysteine.
METHODS： The experiment was carried out in the Department of Neurology, Xijing Hospital, Fourth Military Medical University of Chinese PLA between February anti Augost 2005. The ECV-304 cells were cultured, those of the 4^th-7^th generations at exponential growth phase were taken and inoculated to 96-well plate, the fluid was changed after cell adhesion for 24 hours, and then drug was added. The cells were randomly divided into 3 groups of normal control group, homoeysteine group and flunarizine hy drochloride group. The ECV-304 cells in the normal control group were incubated only in blank RPMI-1640 for 24 hours; those in the homocysteine group were incubated with homocysteine of different concentrations （100, 200, 500 and 1 000 μmol/L） for 24 hours, and those in the flunarizine hydrochloride group were incubated with flunarizine hydrochloride （10 μmmol/L） homocysteine （100, 200, 500 and 1 000 μmol/L） tor 24 hours, 8 wells far each concentration in each group respectively. The absorbane values （wave length of 490 nm） of soluble intercellular adhesion molecule-1 （sICAM-1） and soluble vascular cell adhesion molecule-1 （sVCAM-1） were delected with ELISA.
RESULTS： In the homocysteine group, with the increase of the concentration of homocysteine in culture medium, the absorbance of sICAM-1 was increased from （0.143±0.013） to （0.175±0.006）, which was significantly different from that in the control group （P 〈 0.01）; the absorbance of sVCAM-1 was increased from （0.112±0.008） to （0.147±0.014）, which was significantly different from that in the control group （P 〈 0.01）, The absorbance in the 10 μmmol/L flunarizine hydrochloride ＋100 μmol/L homocysteine group was reduced as compared with that in the 100 μmol/L homocysteine group without significant difference （P 〉 0.05）. The absorbances in the 10 μmmol/L flunarizine hydroehloride ＋200, 500, 1 000 μmol/L humocysteine groups were significantly lower titan those in the eurrespanding homocysteine groups （P 〈 0.01）, indicating thai flunarizine hydrochloride could obviously inhibit the expressions of sICAM-1 and sVCAM-1 induced by homocysteine.
CONCLUSION： It was probable that flunarizine hydrochloride could protect cells from homocysteine injury by down-regulatlng the expression of cellular adhesion molecules induced by homocysteine.
出处
《中国临床康复》
CSCD
北大核心
2006年第14期114-116,共3页
Chinese Journal of Clinical Rehabilitation
