中国血液净化

中国血液净化 ›› 2013, Vol. 12 ›› Issue (09): 496-500.doi: 10.3969/j.issn.1671-4091.2013.09.00

• 基础研究 • 上一篇    下一篇

经外膜缓释雷帕霉素抑制移植血管内膜增生的实验研究

  

  1. 1. 解放军总医院第一附属医院肾内科
    2. 解放军总医院第一附属医院 肾内科
  • 收稿日期:2013-05-08 修回日期:2013-06-08 出版日期:2013-09-12 发布日期:2013-09-12
  • 通讯作者: 李冀军 lijj9536@sina.com E-mail:lijj9536@sina.com

The study of adventitia applied slow-releasing rapamycin inhibiting intima hyperplasia of vascular grafts

  • Received:2013-05-08 Revised:2013-06-08 Online:2013-09-12 Published:2013-09-12

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摘要： 目的 建立兔肾下腹主动脉人造血管移植模型，探讨雷帕霉素抑制人造血管移植后再狭窄的作用机制。方法 健康雄性新西兰大耳白兔18只，体重2.5~3.0 kg，随机分为A、B、C3组，每组6只，构建PTFE人造血管腹主动脉移植模型，血管吻合完成后各组给予不同处理。A组：移植血管不给予任何处理；B组：在移植血管外膜及吻合口周围涂抹配制好的20% pluronic F-127多聚凝胶0.5ml；C组：在移植血管外膜及吻合口周围涂抹携带雷帕霉素0.5mg的pluronic F-127多聚凝胶0.5ml。术后1月获取移植血管，组织形态学方法观察移植血管内膜增生情况，计算机图像分析系统测量移植血管内膜及中膜厚度，并计算内膜增生程度（内膜/中膜），免疫组化SP法检测移植血管α-actin、PCNA和p27kip1的表达。结果 1. 术后1月，A组、B组较C组内膜增生明显（P＜0.05）。2. 增生的血管内膜均可见α-actin阳性表达，类似于血管平滑肌α-actin阳性表达，证实增生的血管内膜于最表层细胞层下表达平滑肌活性。3. PCNA、p27kip1免疫组化观察为胞核着色，阳性细胞呈棕黄色。各组移植血管增生的内膜均可见不同程度的阳性表达。4. α-actin和PCNA的表达，C组较A、B组明显降低（P＜0.05）。p27kip1的表达，C组分别A、B组明显增多（P＜0.05）。A组与B组之间各组数据比较无显著差异（P＞0.05）。结论 1. pluronic F-127多聚凝胶携带雷帕霉素涂抹移植血管外膜可有效抑制移植血管平滑肌细胞增殖。2. 雷帕霉素抑制移植血管再狭窄的机制与其上调移植血管组织中p27kip1的表达、细胞增殖周期受抑有关。

关键词: 人造血管移植, 内膜增生, 雷帕霉素

Abstract: Objective Establish a model of rabbit PTFE artificial vascular grafts on infrarenal abdominal aorta and investigate the mechanism of rapamycin inhibiting restenosis. Methods 18 healthy male New Zealand rabbits weighted 2.5~3.0kg were randomly divided into 3 groups and established models of PTFE artificial vascular graft on abdominal aorta. Group A: the grafts accepted no any management; Group B: 0.5ml 20% pluronic F-127 gel was locally applied to the adventitia of the grafts and anastomoses; Group C: 0.5ml 20% pluronic F-127 gel containing 0.5mg rapamycin was locally applied to the adventitia of the grafts and anastomoses. The grafts were acquired one month after the experiment. Histomorphologic methods were used to detect the intima hyperplasia of the specimen. Electronic imaging system was used to measure the thickness of intima and media of the grafts followed by calculating the degree of intima hyperplasia (thickness of intima/thickness of media). α-actin, PCNA and p27kip1 were investigated by using immunohistochemistry. Data were analyzed by using SPSS13.0 statistics software and then were compiled in the form of（ ）. Results 1. One month after the operation, the intima of Group A and B thickened obviously （P＜0.05）in comparison to C. 2. Result of α-actin immunohistochemistry: the hyperplastic intima was marked obviously, which is consistent with the smooth muscle cells of blood vessels. 3. Result of PCNA and p27kip1 immunohistochemistry: both of them were stained in brown in cell nucleus, and expressed in the thickened intima of every group to study the difference. 4. Compared to A and B, the expression of α-actin and PCNA in C were remarkably inhibited（P＜0.05）, while the expression of p27kip1 was remarkably enhanced（P＜0.05）. No significant deviation was found among A and B（P＞0.05）. Conclusion 1. Smearing rapamycin mixed with pluronic F-127 gel on graft adventitia can effectively inhibit the hyperplasia of vascular smooth muscle cells. 2. The mechanism of rapamycin inhibiting restenosis may be related to increase the expression of p27kip1 and inhibition cell generation cycle.

Key words: artificial vascular graft, intima hyperplasia, rapamycin