血管内皮细胞氧化应激反应对细胞粘附分子表达的影响

Effect of oxidative stress on the expression of cellular surface adhesion molecule

目的 通过对过氧亚硝酸阴离子所诱导的血管内皮细胞的氧化应激反应对内皮细胞表面粘附分子的影响的研究 ,探讨妊娠高血压疾病的发病机制。方法 应用人类脐静脉血管内皮细胞 (humanumbilicalveinendothelialcell,HUVEC)作为体外细胞模型。SIN - 1(3-Morpholinosydnonimine -HCL)作为细胞培养液中过氧亚硝酸阴离子发生器 ,用以刺激培养的血管内皮细胞。以ELISA法检测血管内皮细胞表面粘附分子 ,包括细胞间粘附分子 (intercelluaradhesionmolecule ,ICAM)、血管细胞粘附分子 (vascularadhesionmolecule,VCAM)、P -选择素 (P -Selectin)及E -选择素 (E -Selectin)的表达水平。结果 SIN - 1处理培养之血管内皮细胞 0 5h、 1h ,2h及 4h ,细胞表面粘附分子在经SIN - 1处理 4h后表达水平显著增加 ,呈时间依从性递增的趋势 ;SIN - 1处理培养HUVEC 4h后 ,血管细胞粘附分子 (VCAM)、P -选择素及E -选择素的表达水平较未经SIN - 1处理的HUVEC显著增加 :VCAM (0 115± 0 0 2 2 )vs (0 0 35± 0 0 11) (P <0 0 5 ) ;P -选择素 (0 0 80± 0 0 10 )vs (0 0 34±0 0 0 1) (P <0 0 5 ) ;E -选择素 (0 12 9± 0 0 36 )vs (0 0 37± 0 0 0 4 ) (P <0 0 5 )而ICAM的表达没有显著的变化 (0 2 92± 0 0 1)

Objective The aim of this study was to test whether EC oxidative stress induced by peroxynitite could upregulate EC CAM expression, thus providing a potential strategy in managing the coagulopathy in preeclampsia. Mehtods Human umbilical vein endothelial cell (HUVEC) is exploited as an in vitro cellular model and SIN-1 is used to generate peroxynitite to induce oxidative stress within the HUVEC. Cell adhesion molecules (ICAM, VCAM , P-selectin and E-selectin) were measured by ELISA after 0.5hr,1hr,2hr and 4hr. Ressults Significant increase of endothelial surface adhesion molecules was observed after treatment of SIN-1 for 4hr compared to that before treatment: VCAM(0.115±0.022)vs(0.035±0.011)(P<0.05), P-Selectin(0.080±0.010)vs(0.034±0.001)(P<0.05), E-Selectin(0.129±0.036)vs(0.037±0.004)(P<0.05). Whereas no significant difference was observed in the ICAM expression:(0.292±0.01)vs(0.220±0.019)(P>0.05). At the same time, the endothelial immunostaining for the nitrotyrosine is significantly increased after the cellular challenge by SIN-1.Conclusion Peroxynitrite induced EC oxidative stress produces differential effects on CAM expression.