CAJ | 학술논문

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+雷帕霉素组32只.雷帕霉素按2.0 mg/(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14 d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VEGF表达,实时定量聚合酶链反应(Real-time PCR,染料法)检测VEGF mRNA表达.多个独立样本比较采用Kruskal-Wallis H检验,两样本间的比较经秩转换处理后行one way ANOVA检验,两独立样本比较采用Mann-Whitney U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VEGF mRNA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VEGF mRNA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.
목적 관찰뢰파매소대대서간내담관결혈술후혈관내피생장인자(VEGF) mRNA적영향.방법 120지웅성SD대서수궤분위4조,A조위대조조(가수술조)28지；B조위가수술+뢰파매소조28지；C조위결혈조32지,D조위결혈+뢰파매소조32지.뢰파매소안2.0 mg/(kg·d)위내주입.술전、술후7d급술후14d분별측량실험대서체질량.각실험조우술후제1、3、7천분별처사6지대서,술후14 d처사전부대서.절취간장조직,면역조직화학염색분석관찰간조직내VEGF표체,실시정량취합매련반응(Real-time PCR,염료법)검측VEGF mRNA표체.다개독립양본비교채용Kruskal-Wallis H검험,량양본간적비교경질전환처리후행one way ANOVA검험,량독립양본비교채용Mann-Whitney U검험.결과 대조조급가수술+뢰파매소조미견명현VEGF양성염색；결혈조술후VEGF양성염색명현,주요정위우회관구내혈관내피；결혈조급결혈+뢰파매소조술후VEGFmRNA명현승고,술후3d체도봉치(5.19±0.15),술후1、3、7d,결혈조혈청VEGF mRNA수평고우결혈+뢰파매소조(P＜0.05).결론 뢰파매소억제담관결혈후VEGF mRNA표체,저가능시뢰파매소억제담관결혈후대상성증생적궤제지일.
Objective To investigate the effect of rapamycin on the expression of vascular endothelial 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 ischemia + rapamycin (n =32).In ischemia 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 rapamycin 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,followed by VEGF immunohistochemical 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 ＜ O.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 ischemia 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 duct proliferation in response to ischemia.