替格瑞洛對oxLDL誘導的人臍靜脈內皮細胞ICAM—1表達的影響

劉華　康美尼　王興華

摘 要：目的 探究不同濃度的替格瑞洛對氧化型低密度脂蛋白誘導的人臍靜脈內皮細胞在蛋白及RNA水平表達細胞間黏附分子-1的影響。方法 離體培養人臍靜脈內皮細胞，平均接種于6孔培養板，待細胞生長至融合狀態時取三孔加入ox-LDL（50 μg/ml），再分別加入不同濃度的替格瑞洛（0、20、40 μmol/L）。刺激干預24 h后采用Western Blot法測定ICAM-1的蛋白表達水平。重復實驗，采用Real-time PCR法測定ICAM-1的RNA表達水平。結果 ①oxLDL能明顯上調ICAM-1的表達；②Western Blot蛋白定量結果顯示，oxLDL誘導組ICAM-1的蛋白表達明顯高于對應的非誘導組（P<0.05）；未予替格瑞洛干預組ICAM-1的蛋白表達高于替格瑞洛低濃度組（P<0.05）；替格瑞洛低濃度組ICAM-1的蛋白表達高于替格瑞洛高濃度組（P<0.05）；③Real-time PCR結果與Western Blot蛋白定量結果一致。結論 oxLDL能在RNA水平提高ICAM-1表達，替格瑞洛能在RNA水平抑制ICAM-1的表達，并與濃度正相關。

關鍵詞：替格瑞洛；人臍靜脈內皮細胞；氧化型低密度脂蛋白；細胞間黏附分子-1

中圖分類號：R543 文獻標識碼：A DOI：10.3969/j.issn.1006-1959.2018.22.022

文章編號：1006-1959（2018）22-0081-04

Effect of Ticagrelor on the Expression of ICAM-1 in Human umbilical Vein Endothelial Cells Induced by oxLDL

LIU Hua1，2，KANG Mei-ni2，WANG Xing-hua2，LIANG Xue2，LI Guang-ping2

（1.Department of Cardiology，Cangzhou Central Hospital，Cangzhou 061000，Hebei，China；

2.Institute of Cardiology，Second Hospital，Tianjin Medical University，Tianjin 300211，China）

Abstract：Objective To investigate the effects of different concentrations of ticagrelor on the expression of intercellular adhesion molecule-1 in human umbilical vein endothelial cells induced by oxidized low density lipoprotein at protein and RNA levels.Methods Human umbilical vein endothelial cells were cultured in vitro and seeded on a 6-well culture plate. When the cells were grown to the fusion state， ox-LDL （50 μg/ml） was added to the wells， and different concentrations of ticagrelor were added（0， 20， 40 μmol/L）. The protein expression level of ICAM-1 was determined by Western Blot after 24 h of stimulation. The experiment was repeated and the RNA expression level of ICAM-1 was determined by Real-time PCR. Results ①oxLDL could up-regulate the expression of ICAM-1.②Western Blot protein quantitation results showed that the protein expression of ICAM-1 in oxLDL-induced group was significantly higher than that in the corresponding non-inducing group（P<0.05）；The protein expression of ICAM-1 in the Ticagrelor-free group was higher than that in the tigrisin group （P<0.05）. The protein expression of ICAM-1 in the low-concentration group was higher than that in the Ticagrelor group（P<0.05）； ③Real-time PCR results were consistent with Western Blot protein quantitation results. Conclusion oxLDL can increase the expression of ICAM-1 at the RNA level， and ticagrelor can inhibit the expression of ICAM-1 at the RNA level， and it is positively correlated with the concentration.

Key words：Ticagrelor；Human umbilical vein endothelial cells；Oxidized low density lipoprotein； Intercellular adhesion molecule-1

隨著近年來信號傳導和基因調控水平的研究，動脈粥樣硬化發病機制愈發傾向于炎癥損傷應答學說。粘附分子是炎癥的始動因子和標志物，在動脈粥樣硬化早期炎癥中起著重要的作用。黏附分子-1（ICAM-1）是一種非常重要的具有代表性的粘附因子，在靜息的內皮細胞上呈低水平表達或不表達，但在氧化型低密度脂蛋白（oxLDL）等炎性損傷因子刺激下能迅速上調，導致白細胞停止滾動粘附于內皮細胞，導致一系列心血管病理改變。替格瑞洛（Tigeralor）作為新型抗血小板藥物，在發揮抗血小板作用的同時，同時是否具有抗炎癥損傷作用。

1 材料與方法

1.1材料

1.1.1細胞系 人臍靜脈內皮細胞（human umbilical vein endothelial cells，HUVECs）CRL-1730由天津市心血管疾病研究所贈與，其培養基為含10%胎牛血清（fetal bovine serum，FBS）的DMEM-F12。

1.1.2主要試劑 DMEM-12培養基（GIBC，美國）；oxLDL（北京欣源佳和生物科技有限公司，北京）；……