羅濤 1 , 張強 2 , 張建 1 , 李建新 1
  • 1.首都醫(yī)科大學宣武醫(yī)院血管外科(北京100053);;
  • 2.中國醫(yī)科大學附屬第一臨床醫(yī)院干診病房（沈陽110001）;

目的 　探討腺病毒載體轉(zhuǎn)染單純皰疹病毒胸苷激酶(herpes simplex virus thymidine kinase，HSVtk)基因?qū)σ浦察o脈內(nèi)膜增生的作用。
方法 　用Wistar大鼠建立自體靜脈移植模型。以載有HSVtk(4×109 plaque forming units)基因的腺病毒孵育頸靜脈，將靜脈倒置移植于腎下腹主動脈。從移植術后第3 d開始腹腔注射丙氧鳥苷〔ganciclovir,GCV，60 mg/(kg·d)，2次/d〕共21 d，取出移植靜脈標本后用PCR法檢測HSVtk基因的表達，用原位雜交法檢測基因的轉(zhuǎn)錄； 進行彈力纖維染色及增殖細胞核抗原(PCNA)免疫組化染色，觀察內(nèi)膜增生及平滑肌細胞增殖； 用TUNEL法觀察移植靜脈平滑肌細胞凋亡情況。
結(jié)果 　移植靜脈內(nèi)膜進行性增厚，新生內(nèi)膜中有大量增殖狀態(tài)的平滑肌細胞。移植后第3 d，轉(zhuǎn)染HSVtk基因的靜脈經(jīng)PCR擴增出590 bp陽性條帶，HSVtk基因開始表達mRNA。HSVtk/GCV對靜脈內(nèi)膜增生有明顯抑制作用，而單獨予以HSVtk對內(nèi)膜增生無影響〔(17.2±3.2) μm vs (31.1±2.5) μm, P＜0.05〕。HSVtk/GCV使靜脈中平滑肌細胞增殖指數(shù)下降為(9.1±2.3)%，凋亡細胞指數(shù)升高到(28.7±3.6)%，分別于空白對照組〔(38.7±5.6)%,(18.5±2.6)%〕相比差異均有統(tǒng)計學意義(P＜0.05)。
結(jié)論 　HSVtk/GCV系統(tǒng)通過抑制平滑肌細胞增殖及促進細胞凋亡抑制移植靜脈內(nèi)膜增生。

引用本文： 羅濤,張強,張建,李建新. 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. 中國普外基礎與臨床雜志, 2008, 15(10): 743-747. doi: 復制

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14. 　羅濤， 張強， 張建, 等. 早期炎性反應對移植靜脈中外源基因表達效率的影響 [Ｊ]. 中國普通外科雜志, 2006; 15(5)∶349.
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17. 　Nicholas TW, Read SB, Burrows FJ, et al. Suicide gene therapy with herpes simplex virus thymidine kinase and ganciclovir is enhanced with connexins to improve gap junctions and bystander effects [Ｊ]. Histol Histopathol, 2003; 18(2)∶495.
18. 　羅濤， 張強， 張建, 等. 腺病毒介導胸苷激酶基因?qū)o脈平滑肌細胞增殖及內(nèi)膜增生抑制作用的研究 [Ｊ]. 中國普外基礎與臨床雜志, 2006; 13(3)∶291.
  1. 1. 　Chandiwal A, Balasubramanian V, Baldwin ZK, et al. Gene therapy for the extension of vein graft patency: a review [Ｊ]. Vasc Endovascular Surg, 2005; 39(1)∶1.
  2. 2. 　Appleby CE, Kingston PA. Gene therapy for restenosis——what now, what next? [Ｊ]. Curr Gene Ther, 2004; 4(2)∶153.
  3. 3. 　Izzat MB, West RR, Bryan AJ, et al. Coronary artery bypass surgery: current practice in the United Kingdom [Ｊ]. Br Heart J, 1994; 71(4)∶382.
  4. 4. 　Zou Y, Dietrich H, Hu Y, et al. Mouse model of venous bypass graft arteriosclerosis [Ｊ]. Am J Pathol, 1998; 153(4)∶1301.
  5. 5. 　Stark VK, Hoch JR, Warner TF, et al. Monocyte chemotactic protein-1 expression is associated with the development of vein graft intimal hyperplasia [Ｊ]. Arterioscler Thromb Vasc Biol, 1997; 17(8)∶1614.
  6. 6. 　Greco O, Dachs GU. Gene directed enzyme/prodrug therapy of cancer: historical appraisal and future prospectives [Ｊ]. J Cell Physiol, 2001; 187(1)∶22.
  7. 7. 　TyynelK, Sandmair AM, Turunen M, et al. Adenovirus-mediated herpes simplex virus thymidine kinase gene therapy in BT4C rat glioma model [Ｊ]. Cancer Gene Ther, 2002; 9(11)∶917.
  8. 8. 　Chang MW, Ohno T, Gordon D, et al. Adenovirus-mediated transfer of the herpes simplex virus thymidine kinase gene inhibits vascular smooth muscle cell proliferation and neointima formation following balloon angioplasty of the rat carotid artery [Ｊ]. Mol Med, 1995; 1(2)∶172.
  9. 9. 　Steg PG, Tahlil O, Aubailly N, et al. Reduction of restenosis after angioplasty in an atheromatous rabbit model by suicide gene therapy [Ｊ]. Circulation, 1997; 96(2)∶408.
  10. 10. 　Eslami MH, Gangadharan SP, Sui X, et al. Gene delivery to in situ veins: differential effects of adenovirus and adeno-associated viral vectors [Ｊ]. J Vasc Surg, 2000; 31(6)∶1149.
  11. 11. 　Baker AH, Mehta D, George SJ, et al. Prevention of vein graft failure: potential applications for gene therapy [Ｊ]. Cardiovasc Res, 1997; 35(3)∶442.
  12. 12. 　Ander S, MacLennan M, Bentil S, et al. Pressure-induced vector transport in human saphenous vein [Ｊ]. Ann Biomed Eng, 2005; 33(2)∶202.
  13. 13. 　Rekhter MD, Shah N, Simari RD, et al. Graft permeabilization facilitates gene therapy of transplant arteriosclerosis in a rabbit model [Ｊ]. Circulation, 1998; 98(13)∶1335.
  14. 14. 　羅濤， 張強， 張建, 等. 早期炎性反應對移植靜脈中外源基因表達效率的影響 [Ｊ]. 中國普通外科雜志, 2006; 15(5)∶349.
  15. 15. 　Sandalon Z, Fusenig NE, McCutcheon J, et al. Suicide gene therapy for premalignant disease: a new strategy for the treatment of intraepithelial neoplasia [Ｊ]. Gene Ther, 2001; 8(3)∶232.
  16. 16. 　Burrows FJ, Gore M, Smiley WR, et al. Purified herpes simplex virus thymidine kinase retroviral particles: Ⅲ. Characterization of bystander killing mechanisms in transfected tumor cells [Ｊ]. Cancer Gene Ther, 2002; 9(1)∶87.
  17. 17. 　Nicholas TW, Read SB, Burrows FJ, et al. Suicide gene therapy with herpes simplex virus thymidine kinase and ganciclovir is enhanced with connexins to improve gap junctions and bystander effects [Ｊ]. Histol Histopathol, 2003; 18(2)∶495.
  18. 18. 　羅濤， 張強， 張建, 等. 腺病毒介導胸苷激酶基因?qū)o脈平滑肌細胞增殖及內(nèi)膜增生抑制作用的研究 [Ｊ]. 中國普外基礎與臨床雜志, 2006; 13(3)∶291.