李敬東 1 , 戴毅 1 , 陳開 1 , 鄒琳 2
  • 1.川北醫(yī)學(xué)院附屬醫(yī)院肝膽外科（四川南充 637000）;;
  • 2.重慶醫(yī)科大學(xué)附屬兒童醫(yī)院臨床分子醫(yī)學(xué)中心（重慶 400014）;

目的  探討注射加壓轉(zhuǎn)染的方法對兔動脈壁進行人凝血酶調(diào)節(jié)蛋白（human thrombomodulin, hTM）基因轉(zhuǎn)染的可行性。
方法  健康新西蘭大白兔84只，應(yīng)用抽簽法隨機分為3組： pcDNA3.1/hTM質(zhì)粒組（n=28），pcDNA3.1(+)/neo質(zhì)粒組（n=28）和未轉(zhuǎn)染組（n=28），通過基因轉(zhuǎn)染并建立動脈損傷-阻滯模型，于術(shù)后3、7、14和28 d分別檢測hTM mRNA和蛋白在動脈壁的表達。
結(jié)果  17只實驗動物在手術(shù)當(dāng)天至術(shù)后3 d內(nèi)意外死亡。pcDNA3.1/hTM質(zhì)粒組各時間點的hTM mRNA表達都明顯高于pcDNA3.1(+)/neo質(zhì)粒組和未轉(zhuǎn)染組（P＜0.01）； pcDNA3.1(+)/neo質(zhì)粒組和未轉(zhuǎn)染組組內(nèi)和組間每個時間點的hTM mRNA表達比較，差異均無統(tǒng)計學(xué)意義（P gt;0.05）。hTM蛋白在各組中均有表達，主要位于動脈內(nèi)腔面； pcDNA3.1/hTM質(zhì)粒組各時間點的hTM蛋白表達明顯高于pcDNA3.1(+)/neo質(zhì)粒組和未轉(zhuǎn)染組（P＜0.05）； pcDNA3.1(+)/neo質(zhì)粒組和未轉(zhuǎn)染組的hTM蛋白表達在各個時間點都相對恒定，組內(nèi)和組間比較差異均無統(tǒng)計學(xué)意義（P gt;0.05）。
結(jié)論  用動脈腔內(nèi)加壓法可以有效地對活體動物的動脈壁進行基因轉(zhuǎn)染。

引用本文： 李敬東,戴毅,陳開,鄒琳. 人凝血酶調(diào)節(jié)蛋白基因?qū)ν脛用}壁的轉(zhuǎn)染. 中國普外基礎(chǔ)與臨床雜志, 2009, 16(8): 628-632. doi: 復(fù)制

1. 戴毅， 陳開， 鄒琳， 等. 人血栓調(diào)節(jié)蛋白基因編碼片段的克隆及在真核細胞中的表達 [Ｊ].中國普外基礎(chǔ)與臨床雜志， 2005; 12（6）: 577-580.
2. 戴毅， 陳開， 喬正榮， 等. 凝血酶調(diào)節(jié)蛋白基因?qū)δ氺o脈內(nèi)皮細胞抗凝功能的調(diào)控 [Ｊ]. 中華血液學(xué)雜志， 2006; 27（3）:162-165.
3. Dulak J, Schwarzacher SP, Zwick RH, et al. Effects of local gene transfer of VEGF on neointima formation after balloon injury in hypercholesterolemic rabbits [Ｊ]. Vasc Med, 2005; 10(4): 285-291.
4. J.薩姆布魯克， D.W.拉塞爾主編. 黃培堂主譯. 分子克隆實驗指南 [M]. 上冊. 第3版. 北京: 科學(xué)出版社， 2002: 128-129.
5. Liu Q, Chen ZQ, Bobustuc GC, et al. Local gene transduction of cyclooxygenase-1 increases blood flow in injured atherosclerotic rabbit arteries [Ｊ]. Circulation, 2005; 111(14): 1833-1840.
6. Forte A, Finicelli M, Mattia M, et al. Mesenchymal stem cells effectively reduce surgically induced stenosis in rat carotids [Ｊ]. J Cell Physiol, 2008; 217(3): 789-799.
7. 陳華， 景在平， 包俊敏， 等. RNA干擾對兔血管平滑肌細胞bcl-2基因表達的影響 [Ｊ]. 中國普外基礎(chǔ)與臨床雜志， 2007; 14（1）: 56-58.
8. Nicklin SA, White SJ, Nicol CG, et al. In vitro and in vivo characterisation of endothelial cell selective adenoviral vectors [Ｊ]. J Gene Med, 2004; 6(3): 300-308.
9. Kallenbach K, Salcher R, Heim A, et al. Inhibition of sm-ooth muscle cell migration and neointima formation in vein grafts by overexpression of matrix metalloproteinase-3 [Ｊ]. J Vasc Surg, 2009; 49(3): 750-758.
10. Brunetti-Pierri N, Stapleton GE, Law M, et al. Efficient, long-term hepatic gene transfer using clinically relevant HDAd doses by balloon occlusion catheter delivery in nonhuman primates [Ｊ]. Mol Ther, 2009; 17(2): 327-333.
11. Morishita R. Recent progress in gene therapy for cardiovascular disease [Ｊ]. Circ J, 2002; 66(12): 1077-1086.
12. Li W, Tanaka K, Morioka K, et al. Long-term effect of gene therapy for chronic ischemic myocardium using platelet-derived endothelial cell growth factor in dogs [Ｊ]. J Gene Med, 2008; 10(4): 412-420.
13. 羅濤， 張強， 張建， 等. 轉(zhuǎn)染單純皰疹病毒胸苷激酶基因抑制移植靜脈內(nèi)膜增生 [Ｊ]. 中國普外基礎(chǔ)與臨床雜志， 2008; 15（10）: 743-747.
14. Wu S, Wang X, Guo L, et al. Adenovirus mediated endothelial nitric oxide synthase gene transfer prevents restenosis of vein grafts [Ｊ]. ASAIO J, 2004; 50(3): 272-277.
15. 梁宏立, 朱洪生, 黃忠耀， 等. 兔自體移植靜脈基因轉(zhuǎn)移模型的建立 [Ｊ]. 上海實驗動物科學(xué)， 2000; 20（1）: 3-6.
16. 曲樂豐, 景在平, 曹貴松， 等. 手術(shù)縫線攜載反義基因預(yù)防血管吻合口內(nèi)膜增生的實驗研究 [Ｊ]. 中華普通外科雜志， 2002; 17（1）: 36-37.
17. Wen DZ, Dittman WA, Ye RD, et al. Human thrombomodulin complete cDNA sequence and chromosome localization of the gene [Ｊ]. Biochemistry, 1987; 26(14): 4350-4357.
18. Hirsch T, Spielmann M, Velander P, et al. Insulin-like gr-owth factor-1 gene therapy and cell transplantation in diabetic wounds [Ｊ]. J Gene Med, 2008; 10(11): 1247-1252.
19. Kishida A, Nakashima M, Sakamoto N, et al. Study on complex formation between recombinant human thrombomodulin fragment and thrombin using surface plasmon resonance [Ｊ]. Am J Hematol, 2000; 63(3): 136-140.
20. Yeh HY, Lin JC. Bioactivity and platelet adhesion study of a human thrombomodulin-immobilized nitinol surface [Ｊ]. J Biomater Sci Polym Ed, 2009; 20(5-6): 807-819.
  1. 1. 戴毅， 陳開， 鄒琳， 等. 人血栓調(diào)節(jié)蛋白基因編碼片段的克隆及在真核細胞中的表達 [Ｊ].中國普外基礎(chǔ)與臨床雜志， 2005; 12（6）: 577-580.
  2. 2. 戴毅， 陳開， 喬正榮， 等. 凝血酶調(diào)節(jié)蛋白基因?qū)δ氺o脈內(nèi)皮細胞抗凝功能的調(diào)控 [Ｊ]. 中華血液學(xué)雜志， 2006; 27（3）:162-165.
  3. 3. Dulak J, Schwarzacher SP, Zwick RH, et al. Effects of local gene transfer of VEGF on neointima formation after balloon injury in hypercholesterolemic rabbits [Ｊ]. Vasc Med, 2005; 10(4): 285-291.
  4. 4. J.薩姆布魯克， D.W.拉塞爾主編. 黃培堂主譯. 分子克隆實驗指南 [M]. 上冊. 第3版. 北京: 科學(xué)出版社， 2002: 128-129.
  5. 5. Liu Q, Chen ZQ, Bobustuc GC, et al. Local gene transduction of cyclooxygenase-1 increases blood flow in injured atherosclerotic rabbit arteries [Ｊ]. Circulation, 2005; 111(14): 1833-1840.
  6. 6. Forte A, Finicelli M, Mattia M, et al. Mesenchymal stem cells effectively reduce surgically induced stenosis in rat carotids [Ｊ]. J Cell Physiol, 2008; 217(3): 789-799.
  7. 7. 陳華， 景在平， 包俊敏， 等. RNA干擾對兔血管平滑肌細胞bcl-2基因表達的影響 [Ｊ]. 中國普外基礎(chǔ)與臨床雜志， 2007; 14（1）: 56-58.
  8. 8. Nicklin SA, White SJ, Nicol CG, et al. In vitro and in vivo characterisation of endothelial cell selective adenoviral vectors [Ｊ]. J Gene Med, 2004; 6(3): 300-308.
  9. 9. Kallenbach K, Salcher R, Heim A, et al. Inhibition of sm-ooth muscle cell migration and neointima formation in vein grafts by overexpression of matrix metalloproteinase-3 [Ｊ]. J Vasc Surg, 2009; 49(3): 750-758.
  10. 10. Brunetti-Pierri N, Stapleton GE, Law M, et al. Efficient, long-term hepatic gene transfer using clinically relevant HDAd doses by balloon occlusion catheter delivery in nonhuman primates [Ｊ]. Mol Ther, 2009; 17(2): 327-333.
  11. 11. Morishita R. Recent progress in gene therapy for cardiovascular disease [Ｊ]. Circ J, 2002; 66(12): 1077-1086.
  12. 12. Li W, Tanaka K, Morioka K, et al. Long-term effect of gene therapy for chronic ischemic myocardium using platelet-derived endothelial cell growth factor in dogs [Ｊ]. J Gene Med, 2008; 10(4): 412-420.
  13. 13. 羅濤， 張強， 張建， 等. 轉(zhuǎn)染單純皰疹病毒胸苷激酶基因抑制移植靜脈內(nèi)膜增生 [Ｊ]. 中國普外基礎(chǔ)與臨床雜志， 2008; 15（10）: 743-747.
  14. 14. Wu S, Wang X, Guo L, et al. Adenovirus mediated endothelial nitric oxide synthase gene transfer prevents restenosis of vein grafts [Ｊ]. ASAIO J, 2004; 50(3): 272-277.
  15. 15. 梁宏立, 朱洪生, 黃忠耀， 等. 兔自體移植靜脈基因轉(zhuǎn)移模型的建立 [Ｊ]. 上海實驗動物科學(xué)， 2000; 20（1）: 3-6.
  16. 16. 曲樂豐, 景在平, 曹貴松， 等. 手術(shù)縫線攜載反義基因預(yù)防血管吻合口內(nèi)膜增生的實驗研究 [Ｊ]. 中華普通外科雜志， 2002; 17（1）: 36-37.
  17. 17. Wen DZ, Dittman WA, Ye RD, et al. Human thrombomodulin complete cDNA sequence and chromosome localization of the gene [Ｊ]. Biochemistry, 1987; 26(14): 4350-4357.
  18. 18. Hirsch T, Spielmann M, Velander P, et al. Insulin-like gr-owth factor-1 gene therapy and cell transplantation in diabetic wounds [Ｊ]. J Gene Med, 2008; 10(11): 1247-1252.
  19. 19. Kishida A, Nakashima M, Sakamoto N, et al. Study on complex formation between recombinant human thrombomodulin fragment and thrombin using surface plasmon resonance [Ｊ]. Am J Hematol, 2000; 63(3): 136-140.
  20. 20. Yeh HY, Lin JC. Bioactivity and platelet adhesion study of a human thrombomodulin-immobilized nitinol surface [Ｊ]. J Biomater Sci Polym Ed, 2009; 20(5-6): 807-819.