【摘要】目的觀察供肝轉染IL-10后基因表達情況。
方法應用改良“二袖套法”行Lewis到BN大鼠肝移植11例,然后分為對照組3例; 空載體轉染組4例,術中供肝冷保存期門靜脈注射Lipofectamine 2000-pCR3.1空載體質粒復合物,保存45 min后行肝移植; 重組IL-10(rIL-10)轉染組4例,術中供肝冷保存期門靜脈注射Lipofectamine 2000-pCR3.1 rIL-10復合物,保存45 min后行肝移植。術后第6天處死全部大鼠,取血清檢測IL-10水平,另取肝組織行免疫組化染色和RT-PCR檢測肝細胞IL-10表達水平。
結果rIL-10轉染組血清IL-10水平明顯升高,肝上下腔靜脈IL-10水平可達(639.27±67.11) pg/ml,是肝下下腔靜脈IL-10水平的近1.4倍(P=0.024); 免疫組化染色結果示肝細胞胞槳呈棕黃色或深棕色,而其他兩組肝細胞著色輕微或不明顯。肝組織RT-PCR顯示,IL-10 mRNA的表達在rIL-10轉染組明顯處于高水平(P=0.000)。
結論脂質體介導,體外冷保存期經門靜脈途徑進行供肝轉染IL-10,可以使IL-10在肝臟獲得較高水平的表達。
引用本文: 張棟,高東宸,張忠濤,王宇. 大鼠肝移植中脂質體介導IL-10基因轉染供肝的實驗研究. 中國普外基礎與臨床雜志, 2005, 12(2): 123-126. doi: 復制
版權信息: ?四川大學華西醫(yī)院華西期刊社《中國普外基礎與臨床雜志》版權所有,未經授權不得轉載、改編
1. | Kamada N, Calne RY. Orthotopic liver transplantation in the rat: Technique using cuff for portal vein anastomosis and biliary drainage [J]. Transplantation, 1979; 28(1)∶47. |
2. | Kamada N, Sumimoto R, Kaneda K. The value of hepatic artery reconstruction as a technique in rat liver transplantation [J]. Surgery, 1992; 111(2)195. |
3. | Sen L, Hong YS, Luo H, et al. Efficiency, efficacy, and adverse effects of adenovirus vs. liposomemediated gene therapy in cardiac allografts [J]. Am J Physiol Heart Circ Physiol, 2001; 281(3)H1433. |
4. | Shinozaki K, Yahata H, Tanji H, et al. Allograft transduction of IL10 prolongs survival following orthotopic liver transplantation [J]. Gene Ther, 1999; 6(5)816. |
5. | Herweijer H, Wolff JA. Progress and prospects: naked DNA gene transfer and therapy [J]. Gene Ther, 2003; 10(6)453. |
6. | Kren BT, Chowdhury NR, Chowdhury JR, et al. Gene therapy as an alternative to liver transplantation [J]. Liver Transpl, 2002; 8(12)1089. |
7. | Itano H, Mora BN, Zhang W, et al. Lipidmediated ex vivo gene transfer of viral interleukin 10 in rat lung allotransplantation [J]. J Thorac Cardiovasc Surg, 2001; 122(1)29. |
- 1. Kamada N, Calne RY. Orthotopic liver transplantation in the rat: Technique using cuff for portal vein anastomosis and biliary drainage [J]. Transplantation, 1979; 28(1)∶47.
- 2. Kamada N, Sumimoto R, Kaneda K. The value of hepatic artery reconstruction as a technique in rat liver transplantation [J]. Surgery, 1992; 111(2)195.
- 3. Sen L, Hong YS, Luo H, et al. Efficiency, efficacy, and adverse effects of adenovirus vs. liposomemediated gene therapy in cardiac allografts [J]. Am J Physiol Heart Circ Physiol, 2001; 281(3)H1433.
- 4. Shinozaki K, Yahata H, Tanji H, et al. Allograft transduction of IL10 prolongs survival following orthotopic liver transplantation [J]. Gene Ther, 1999; 6(5)816.
- 5. Herweijer H, Wolff JA. Progress and prospects: naked DNA gene transfer and therapy [J]. Gene Ther, 2003; 10(6)453.
- 6. Kren BT, Chowdhury NR, Chowdhury JR, et al. Gene therapy as an alternative to liver transplantation [J]. Liver Transpl, 2002; 8(12)1089.
- 7. Itano H, Mora BN, Zhang W, et al. Lipidmediated ex vivo gene transfer of viral interleukin 10 in rat lung allotransplantation [J]. J Thorac Cardiovasc Surg, 2001; 122(1)29.