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(dòng)脈。從移植術(shù)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(biāo)本后用PCR法檢測(cè)HSVtk基因的表達(dá),用原位雜交法檢測(cè)基因的轉(zhuǎn)錄; 進(jìn)行彈力纖維染色及增殖細(xì)胞核抗原(PCNA)免疫組化染色,觀察內(nèi)膜增生及平滑肌細(xì)胞增殖; 用TUNEL法觀察移植靜脈平滑肌細(xì)胞凋亡情況。
結(jié)果 移植靜脈內(nèi)膜進(jìn)行性增厚,新生內(nèi)膜中有大量增殖狀態(tài)的平滑肌細(xì)胞。移植后第3 d,轉(zhuǎn)染HSVtk基因的靜脈經(jīng)PCR擴(kuò)增出590 bp陽性條帶,HSVtk基因開始表達(dá)mRNA。HSVtk/GCV對(duì)靜脈內(nèi)膜增生有明顯抑制作用,而單獨(dú)予以HSVtk對(duì)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(xì)胞增殖指數(shù)下降為(9.1±2.3)%,凋亡細(xì)胞指數(shù)升高到(28.7±3.6)%,分別于空白對(duì)照組〔(38.7±5.6)%,(18.5±2.6)%〕相比差異均有統(tǒng)計(jì)學(xué)意義(P<0.05)。
結(jié)論 HSVtk/GCV系統(tǒng)通過抑制平滑肌細(xì)胞增殖及促進(jìn)細(xì)胞凋亡抑制移植靜脈內(nèi)膜增生。
引用本文: 羅濤,張強(qiáng),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(chǔ)與臨床雜志, 2008, 15(10): 743-747. doi: 復(fù)制
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14. | 羅濤, 張強(qiáng), 張建, 等. 早期炎性反應(yīng)對(duì)移植靜脈中外源基因表達(dá)效率的影響 [J]. 中國普通外科雜志, 2006; 15(5)∶349. |
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- 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. Appleby CE, Kingston PA. Gene therapy for restenosis——what now, what next? [J]. Curr Gene Ther, 2004; 4(2)∶153.
- 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. Zou Y, Dietrich H, Hu Y, et al. Mouse model of venous bypass graft arteriosclerosis [J]. Am J Pathol, 1998; 153(4)∶1301.
- 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. 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. TyynelK, 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. 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. 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. 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. 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. 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. 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. 羅濤, 張強(qiáng), 張建, 等. 早期炎性反應(yīng)對(duì)移植靜脈中外源基因表達(dá)效率的影響 [J]. 中國普通外科雜志, 2006; 15(5)∶349.
- 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. 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. 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(qiáng), 張建, 等. 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 [J]. 中國普外基礎(chǔ)與臨床雜志, 2006; 13(3)∶291.