经外膜缓释雷帕霉素抑制移植血管内膜增生的实验研究  

The inhibition of intimal hyperplasia in vascular grafts by slow-releasing rapamycin applied on the adventitia: an experimental study

作　　者：毕铭霞[1] 朱彬[2] 王小平[2] 李冀军[1] 

机构地区：[1]解放军总医院第一附属医院肾内科,北京100048 [2]山东大学附属济南市中心医院肾脏病/血液净化中心,济南250013

出　　处：《中国血液净化》2013年第9期496-500,共5页Chinese Journal of Blood Purification

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,体质量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。③PCNA、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。④α-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①pluronic 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。Objective To establish a model of rabbit PTFE artificial vascular grafts at infrarenal abdominal aorta and to investigate the mechanism of re-stenosis inhibition by rapamycin.Methods Eighteen healthy New Zealand male rabbits weighted 2.5~3.0 kg were randomly divided into 3 groups to establish models of PTFE artificial vascular graft at abdominal aorta.Group A： the grafts accepted no management;group B： 0.5ml of 20% pluronic F-127 gel was locally applied on adventitia and anastomosis site of the graft;group C： 0.5ml 20% pluronic F-127 gel containing 0.5mg rapamycin was locally applied on adventitia and anastomosis site of the graft.The grafts were acquired after one month.Histomorphological method was used to detect the intimal 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 intimal hyperplasia（thickness of intima/thickness of media）.α-actin,PCNA and p27kip1were investigated by using immunohistochemistry.Data were analyzed by using SPSS13.0 statistics software,and were expressed as mean ± SD.Results One month after the operation,the intima in group A and B thickened obviously as compared with that in group C（P0.05）.Immunohistochemically,α-actin expressed in the hyperplastic intima,similar to the α-actin expression in smooth muscle cells of blood vessels.Immunohistochemistry for PCNA and p27kip1demonstrated that they were stained in cell nuclei,and were expressed in the thickened intima in grafts from the 3 groups.The expressions of α-actin and PCNA were lower in group C than in groups A and B（P0.05）,while the expression of p27kip1was higher in group C than in groups A and B（P0.05）.No significant differences in the above parameters were found between groups A and B（P0.05）.Conclusion External application of rapamycin mixed with pluronic F-127 gel on graft adventitia can effectively inhibit the hyperplasia of vascular smooth muscle cells.The mechanism of re-stenosis inhibition

关 键 词：人造血管移植 内膜增生 雷帕霉素 

分 类 号：R318.16[医药卫生—生物医学工程]