雷帕霉素对大鼠胆管缺血术后血管内皮生长因子表达的影响  

Effect of rapamycin on expression of vascular endothelial growth factor mRNA after biliary ische-mia in rats

作　　者：吴田田[1] 李虎城[1] 黄辉[1] 张维[1] 赵军[1] 杨广顺[1] 

机构地区：[1]解放军309医院器官移植中心肝胆三科,北京100091

出　　处：《中华实验外科杂志》2012年第11期2125-2127,F0003,共4页Chinese Journal of Experimental Surgery

摘　　要：目的观察雷帕霉素对大鼠肝内胆管缺血术后血管内皮生长因子（VEGF）mRNA的影响。方法120只雄性SD大鼠随机分为4组，A组为对照组（假手术组）28只；B组为假手术＋雷帕霉素组28只；C组为缺血组32只，D组为缺血十雷帕霉素组32只。雷帕霉素按2．0mg／（kg·d）胃内注入。术前、术后7d及术后14d分别测量实验大鼠体质量。各实验组于术后第1、3、7天分别处死6只大鼠，术后14d处死全部大鼠。切取肝脏组织，免疫组织化学染色分析观察肝组织内VEEGF表达，实时定量聚合酶链反应（Real-timePCR，染料法）检测VEGFmRNA表达。多个独立样本比较采用KmskM-WallisH检验，两样本间的比较经秩转换处理后行onewayANOVA检验，两独立样本比较采用Mann-WhitneyU检验。结果对照组及假手术＋雷帕霉素组未见明显VEGF阳性染色；缺血组术后VEGF阳性染色明显，主要定位于汇管区内血管内皮；缺血组及缺血＋雷帕霉素组术后VEGFmRNA明显升高，术后3d达到峰值（5．19±0．15），术后1、3、7d，缺血组血清VEGFmRNA水平高于缺血＋雷帕霉素组（P〈0．05）。结论雷帕霉素抑制胆管缺血后VEGFmRNA表达，这可能是雷帕霉素抑制胆管缺血后代偿性增生的机制之一。Objective To investigate the effect of rapamycin on the expression of vascular endo- thelial growth factor （VEGF） mRNA after deprivation of biliary blood supply. Methods Male SD rats were randomly assigned into 4 groups ： sham （ n = 28 ） , sham ＋ rapamycin （ n = 28 ） , ischemia （ n = 32 ） and isehemia ＋ rapamycin （ n = 32）. In isehemia groups, complete deprivation of bile duct arterial supply was performed, and in sham groups, open-close operation was carried out. Daily intake of rapamycin （2 mg/kg） was given in rapamyein treated groups with the same volume of saline in non-rapamycin treated groups. Body weight was measured before operation, and on the postoperative day （POD） 7 and 14. Six rats were sacrificed on POD 1, 3 and 7, and the rest on POD 14. Fresh liver tissues were obtained, fol- lowed by VEGF immunohistoehemieal stain and real-time polymerase chain reaction （ Real-time PCR） （ dye method）. Kruskal-Wallis H test was used for comparison of multiple independent samples and the following pairwise comparisons were performed by rank transformation and ONE WAY ANOVA. Statistic difference was considered when P 〈 0. 05. Results In ischemia group, obvious VEGF positive staining was found within portal areas, which mainly located at vascular endothelium. In sham or sham ＋ rapamycin groups, less obvious positive staining was found right after surgey, and VEGF mRNA was increased sharply in is- chemia group and reached the peak level on POD 3 （5.19 ±0. 15）. On POD 1, 3 and 7, VEGF mRNA expression level was significantly lower in ischemia ＋ rapamycin group than in ischemia group （ P 〈 0. 05 ）. Conclusion Rapamycin inhibits VEGF mRNA expression, which provides explanation to negative effect of rapamycin on adaptive bile duet proliferation in response to ischemia.

关 键 词：雷帕霉素 胆管缺血 血管内皮生长因子 

分 类 号：R657.4[医药卫生—外科学]