肝部分切除术对大鼠门静脉途径肝细胞转基因效率的影响

目的 :探讨肝部分切除术对大鼠肝细胞门静脉途径转基因效率的影响。方法 :Wistar大鼠分 4组 ,每组 5只 :A1组 :30 %切肝并于门静脉注射脂质体—质粒 DNA混合物 (lipofectamine- p EGFP- N1DNA complexes,lipoplexes) ;A2组 :30 %切肝并于门静脉注射生理盐水 (NS) ;B1组 :70 %切肝并于门静脉注射 lipoplexes;B2组 :70 %切肝并于门静脉注射 NS。转染后 15 d均以胶原酶消化、Percoll液梯度离心法获取纯化肝细胞悬液 ;以 NS组为对照并以 GFP为荧光标记物 ,用流式细胞仪 (flow cytometry,FCM)分析各实验组肝细胞的转染率。结果 :A1组和 B1组的平均转染率分别为 :(1.12± 0 .5 0 ) %、(3.91± 1.6 8) % ;B1组明显高于 A1组 (P<0 .0 5 )。结论 :与 30 %肝部分切除术相比 ,70%切肝可显著提高大鼠肝细胞门静脉途径转基因的效率

Effect of partial hepatectomy on hepatocyte transfection efficiency via rat portal vein

Objective:To study the Effect of partial hepatectomy(PH) on hepatocyte transfection efficiency via rat portal vein.Methods:Twenty Wistar rats were divided into 4 groups(5 rats in each group).Regenerating liver model of rat was established by PH(30 % of 70 %)and isolated hepatic perfusion of lipofectamine-PEGFP-N 1 DNA complexes(lipoplexes,Group A1 and B1 with 30 % and 70 % PH respectively)or normal saline(NS,Group A2 and B2 with 30 % and 70 % PH respectively)via portal vein was carried out 24 h postoperation.Purified hepatocytes were obtained by collagenase digestion followed by Percoll gradient centrifugation 15 d posttransfection.The hepatocytes were analyzed and singled out by flow cytometry(FCM).Results:The mean transfection efficiency analyzed by FCM were (1.12±0.50) % (group A1) and (3.91±1.68) % (group B1)respectively.The efficiency of group B1 was significantly higher than group A1(P<0.05).Conclusion:Compared with 30 % PH,70 % PH could obviously increase hepatocyte transgenic efficiency via rat portal vein.