• 1.首都醫(yī)科大學(xué)血管外科研究所、首都醫(yī)科大學(xué)宣武醫(yī)院血管外科（北京100053）;;
  • 2.中國(guó)醫(yī)科大學(xué)附屬第一臨床醫(yī)院血管外科(沈陽110001);

目的探討以腺病毒為載體介導(dǎo)胸苷激酶（tk）基因?qū)o脈平滑肌細(xì)胞增殖及內(nèi)膜增生的抑制作用。
方法腺病毒載體攜tk及β半乳糖苷酶（lacZ）基因轉(zhuǎn)染人的大隱靜脈平滑肌細(xì)胞及大隱靜脈片,以不同濃度的更昔洛韋（GCV）培養(yǎng)。通過Xgal染色觀察標(biāo)記基因lacZ的表達(dá)效率。將轉(zhuǎn)染tk基因陽性與tk基因陰性的平滑肌細(xì)胞以不同的比例混合培養(yǎng)觀察旁觀者效應(yīng)。將轉(zhuǎn)染tk基因的靜脈片在含有GCV的培養(yǎng)液中培養(yǎng)14 d，取標(biāo)本進(jìn)行HE、VG染色，輔以計(jì)算機(jī)圖像分析，觀察內(nèi)膜增生情況。
結(jié)果腺病毒載體能有效地轉(zhuǎn)染培養(yǎng)的平滑肌細(xì)胞和靜脈片中的平滑肌細(xì)胞，外源基因表達(dá)持續(xù)14 d。平滑肌細(xì)胞增殖受抑制程度與GCV的濃度和tk表達(dá)的水平呈正相關(guān)。轉(zhuǎn)染lacZ的平滑肌細(xì)胞則不被GCV抑制?；旌霞?xì)胞培養(yǎng)（tk陽性及tk陰性細(xì)胞混合），當(dāng)10%的細(xì)胞表達(dá)tk基因時(shí)，細(xì)胞生存率下降了55%。在靜脈片培養(yǎng)實(shí)驗(yàn)中，tk/GCV組與對(duì)照組（lacZ/GCV和tk/GCV-）比較，前者內(nèi)膜增生60%受到抑制。
結(jié)論tk基因合并GCV治療具有抑制平滑肌細(xì)胞增殖和移植靜脈內(nèi)膜增生的作用, tk基因治療具有旁觀者效應(yīng)。

引用本文： 羅濤,張強(qiáng),張建,李建新,谷涌泉,俞恒錫,汪忠鎬. 腺病毒介導(dǎo)胸苷激酶基因?qū)o脈平滑肌細(xì)胞增殖及內(nèi)膜增生抑制作用的研究. 中國(guó)普外基礎(chǔ)與臨床雜志, 2006, 13(3): 291-294. doi: 復(fù)制

版權(quán)信息： ?四川大學(xué)華西醫(yī)院華西期刊社《中國(guó)普外基礎(chǔ)與臨床雜志》版權(quán)所有，未經(jīng)授權(quán)不得轉(zhuǎn)載、改編

1. Davies MG, Hagen PO. Pathophysiology of vein graft failure: a review [Ｊ]. Eur J Vasc Endovasc Surg， 1995; 9(1)∶7.
2. Ip JH, Fuster V, Badimon L, et al. Syndromes of accelerated atherosclerosis: role of vascular injury and smooth muscle cell proliferation [Ｊ]. J Am Coll Cardiol， 1990; 15(7)∶1667.
3. Bulkley BH, Hutchins GM. Accelerated “atherosclerosis”. A morphologic study of 97 saphenous vein coronary artery bypass grafts [Ｊ]. Circulation， 1977; 55(1)∶163.
4. Guzman RJ, Hirschowitz EA, Brody SL, et al. In vivo suppression of injuryinduced vascular smooth muscle cell accumulation using adenovirusmediated transfer of the herpes simplex virus thymidine kinase gene [Ｊ]. Proc Natl Acad Sci USA， 1994; 91(22)∶10732.
5. 羅濤， 張強(qiáng)， 刁彥鵬， 等. 腺病毒載體轉(zhuǎn)染人大隱靜脈平滑肌細(xì)胞效率的研究 [Ｊ]. 中華實(shí)驗(yàn)外科雜志, 2003； 20（3）∶209.
6. Francis SC, Raizada MK, Mangi AA, et al. Genetic targeting for cardiovascular therapeutics: are we near the summit or just beginning the climb? [Ｊ]. Physiol Genomics, 2001; 7(2)∶79.
7. van Dillen IJ, Mulder NH, Vaalburg W, et al. Influence of the bystander effect on HSVtk/GCV gene therapy. A review [Ｊ]. Curr Gene Ther, 2002; 2(3)∶307.
8. Channon KM, Fulton GJ, Gray JL, et al. Efficient adenoviral gene transfer to early venous bypass grafts: comparison with native vessels [Ｊ]. Cardiovasc Res， 1997; 35(3)∶505.
9. Borrelli E, Heyman R, Hsi M, et al. Targeting of an inducible toxic phenotype in animal cells [Ｊ]. Proc Natl Acad Sci USA， 1988; 85(20)∶7572.
10. Robe PA, Jolois O, N’Guyen M, et al. Modulation of the HSVTK/ganciclovir bystander effect by nbutyrate in glioblastoma: correlation with gapjunction intercellular communication [Ｊ]. Int J Oncol， 2004; 25(1)∶187.
11. Soyombo AA, Angelini GD, Bryan AJ, et al. Intimal proliferation in an organ culture of human saphenous vein [Ｊ]. Am J Pathol， 1990; 137(6)∶1401.
12. Ojha M. Wall shear stress temporal gradient and anastomotic intimal hyperplasia [Ｊ]. Circ Res， 1994; 74(6)∶1227.
  1. 1. Davies MG, Hagen PO. Pathophysiology of vein graft failure: a review [Ｊ]. Eur J Vasc Endovasc Surg， 1995; 9(1)∶7.
  2. 2. Ip JH, Fuster V, Badimon L, et al. Syndromes of accelerated atherosclerosis: role of vascular injury and smooth muscle cell proliferation [Ｊ]. J Am Coll Cardiol， 1990; 15(7)∶1667.
  3. 3. Bulkley BH, Hutchins GM. Accelerated “atherosclerosis”. A morphologic study of 97 saphenous vein coronary artery bypass grafts [Ｊ]. Circulation， 1977; 55(1)∶163.
  4. 4. Guzman RJ, Hirschowitz EA, Brody SL, et al. In vivo suppression of injuryinduced vascular smooth muscle cell accumulation using adenovirusmediated transfer of the herpes simplex virus thymidine kinase gene [Ｊ]. Proc Natl Acad Sci USA， 1994; 91(22)∶10732.
  5. 5. 羅濤， 張強(qiáng)， 刁彥鵬， 等. 腺病毒載體轉(zhuǎn)染人大隱靜脈平滑肌細(xì)胞效率的研究 [Ｊ]. 中華實(shí)驗(yàn)外科雜志, 2003； 20（3）∶209.
  6. 6. Francis SC, Raizada MK, Mangi AA, et al. Genetic targeting for cardiovascular therapeutics: are we near the summit or just beginning the climb? [Ｊ]. Physiol Genomics, 2001; 7(2)∶79.
  7. 7. van Dillen IJ, Mulder NH, Vaalburg W, et al. Influence of the bystander effect on HSVtk/GCV gene therapy. A review [Ｊ]. Curr Gene Ther, 2002; 2(3)∶307.
  8. 8. Channon KM, Fulton GJ, Gray JL, et al. Efficient adenoviral gene transfer to early venous bypass grafts: comparison with native vessels [Ｊ]. Cardiovasc Res， 1997; 35(3)∶505.
  9. 9. Borrelli E, Heyman R, Hsi M, et al. Targeting of an inducible toxic phenotype in animal cells [Ｊ]. Proc Natl Acad Sci USA， 1988; 85(20)∶7572.
  10. 10. Robe PA, Jolois O, N’Guyen M, et al. Modulation of the HSVTK/ganciclovir bystander effect by nbutyrate in glioblastoma: correlation with gapjunction intercellular communication [Ｊ]. Int J Oncol， 2004; 25(1)∶187.
  11. 11. Soyombo AA, Angelini GD, Bryan AJ, et al. Intimal proliferation in an organ culture of human saphenous vein [Ｊ]. Am J Pathol， 1990; 137(6)∶1401.
  12. 12. Ojha M. Wall shear stress temporal gradient and anastomotic intimal hyperplasia [Ｊ]. Circ Res， 1994; 74(6)∶1227.