天津医药

天津医药 ›› 2015, Vol. 43 ›› Issue (12): 1401-1404.doi: 10.11958/j.issn.0253-9896.2015.12.015

• 实验研究 • 上一篇    下一篇

雷帕霉素洗脱支架损伤血管内皮的作用机制探讨

袁玲，聂卫，高萍，陈斌，刘伟伟，崔晓雪   

  1. 天津市医药科学研究所（邮编300020）
  • 收稿日期:2015-05-18 修回日期:2015-09-14 出版日期:2015-12-15 发布日期:2015-12-11
  • 作者简介:袁玲（1979），女，助理研究员，硕士，主要从事药物毒理、病理，医疗器械安全性评价方面研究
  • 基金资助:
    天津市卫生局科技基金项目（2013KY37）

The role of rapamycin-eluted stent in vascular endothelial injury

YUAN Ling，NIE Wei，GAO Ping，CHEN Bin，LIU Weiwei，CUI Xiaoxue   

  1. Tianjin Institute of Medical Science，Tianjin 300020，China
  • Received:2015-05-18 Revised:2015-09-14 Published:2015-12-15 Online:2015-12-11

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摘要： 目的探讨雷帕霉素洗脱支架损伤血管内皮的作用机制。方法（1）对兔行局部肌肉注射雷帕霉素（雷帕霉素组），模拟雷帕霉素洗脱支架安全性评价中的肌肉植入，同时设正常对照组和丙酮对照组。观察各组形态变化及测定匀浆液钙含量。（2）采用雷帕霉素0.1、1、10 及100 μg/L 处理人脐静脉内皮细胞（HUVEC）48 h 及1 μg/L 雷帕霉素处理HUVEC 6、12、24 及48 h，采用MTT 法检测细胞存活率，分析细胞存活率与药物浓度及处理时间的关系。（3）HUVEC 分设正常对照组、1 μg/L 雷帕霉素组。48 h 后HE 染色观察细胞形态变化，用NO 检测试剂盒测定细胞培养液NO 含量，采用Fluo-3/AM 荧光探针标记细胞内Ca2+。结果（1）雷帕霉素组出现支架植入类似的蓝染表现，雷帕霉素组较正常对照组和丙酮对照组钙含量升高（P < 0.05）。（2）雷帕霉素处理HUVEC 后，细胞存活率随浓度增加和时间延长均呈逐渐降低趋势（P < 0.01）。（3）雷帕霉素组细胞胞浆出现空泡、核固缩及核碎裂，与正常对照组比较，细胞培养液NO 含量降低，细胞内游离Ca2+含量增高（P < 0.05）。结论雷帕霉素洗脱支架中抗增殖药雷帕霉素对血管内皮有损伤作用，其机制可能与引起细胞内钙升高有关。

关键词: 西罗莫司, 内皮, 血管, 药物洗脱支架, 脐静脉, 钙, 疾病模型, 动物

Abstract: Objective To explore the mechanism of rapamycin -eluted stent in vascular endothelial injury. Meth⁃ ods （1）Rapamycin (rapamycin group) was injected to rabbit dorsum muscle to simulate rapamycin-eluted stent implanta⁃ tion into muscles. Control group and acetone control group were established at the same time. Morphological change in mus⁃ cle was observed and serum calcium levels were measured after rapamycin injection.（2）HUVECs were incubated with 0.1, 1, 10 and 100 μg/L rapamycin for 48 h respectively or with 1 μg/L rapamycin for 6, 12, 24 and 48 h respectively．Cell via⁃ bility was examined by MTT and its relationship with drug concentration and treatment time were analyzed.（3）HUVECs were divided into control group and 1 μg/L rapamycin group. After 48 h，morphological changes of HUVECs were assessed by HE staining，the production of nitric oxide was examined by Nitric Oxide Assay Kit and the intracellular calcium ion con⁃ centration was tagged with Fluo-3/AM. Results (1) Organizational morphology in local muscle with rapamycin injection represent stent implantation of rabbit，and calcium content in local muscle increased significantly in rapamycin group com⁃ pared with nomal control group and acetone control group（P < 0.05）. (2) Cell survival rate decreased significantly upon ad⁃ ministration of rapamycin in both concentration and time dependent manner（P < 0.01）. (3) In rapamycin group, cytoplasm vacuoles, nucleus pycnosis and nuclear fragmentation were observed；compared with control group，the levels of intracellular free Ca2+ increased while the levels of nitric oxide was reduced. Conclusion Rapamycin treatment lead of injury to vascular endothelial cells which might through up-regulating intracellular Ca2+ level.

Key words: Sirolimus, endothelium, vascular, drug-eluting stents, umbilical veins, calcium, disease models, animal