早期炎性反应对移植静脉中外源基因表达效率的影响
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罗涛

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Effect of early inflammation on the efficacy of gene transfer to vein graft
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    摘要:

    目的: 探讨早期炎性反应对移植静脉中腺病毒载体转染的外源基因表达效率的影响。方法:通过颈外静脉将携lacZ基因的腺病毒载体注入大鼠的颈总静脉中,使其自然充盈。室温下孵育30min后,将转基因静脉间置移植于颈总动脉。在静脉转基因后3,14,21d取材。经X-Gal(5-溴-4-氯-3-吲哚-D-半乳糖苷)染色检测静脉中标记基因的表达,测定β-半乳糖酶的活性。常规免疫组化染色观察静脉中细胞内黏附分子-1(ICAM-1)及血管细胞黏附分子-1(VCAM-1)的表达。结果:外源基因在体表达14d时β-半乳糖酶活性下降,至21d时活性近消失。静脉移植后3d,VCAM-1和ICAM-1表达上调,白细胞浸润,内皮细胞脱落,内皮层不完整。而未进行移植的转基因静脉,内皮细胞无VCAM-1及ICAM-1的表达,血管壁无炎性细胞浸润,内皮细胞完整。在移植静脉中,阳性内皮细胞数比未移植静脉明显减少(P<0.05),平滑肌细胞数则与未移植静脉相比无明显差别。结论:移植静脉早期炎性反应造成的内皮细胞损伤与转基因表达迅速下降有关。

    Abstract:

    Abstract:Objective:To investigate the effect of early inflamation on the efficacy of gene(LacZ gene) transfer to vein graft by adenovirus vector.Methods:The liquid containing adenovirus vector was infused through external carotid vein and the segment of carotid vein was resected. The veins were inflated naturally. After 30min incubation, the veins were transplanted into carotid artery in experimental group, and the circulation was restored in control group. The veins with transferred gene were collected on day 3,day 14 and day 21. The mark gene expression in the veins was studied with X-Gal stain and activities of β-galactase were measured. ICAM-1 and VCAM-1 expressions were observed by immunohistochemistry stain in transgenic veins.Results:The activities of β-galactase in mark gene expressed for 14 days was decreased and in 21 day nearly disappearred. VCAM-1 and ICAM-1 expression upregulated after day 3 in transplanted veins and there was leucocyte infiltration and shedding of endothelial cells with endothelial disruption. On the other hand, there was no expression of VCAM-1 and ICAM-1 in control group veins and integrity of endothelium was intact. The positive ECs expressed mark gene was obviously reduced in transplanted veins compared to control group. But the number of positive SMCs were not significently different between to two groups.Conclusions:The inflammation of vein grafts in early stage of transplantation may be the cause of endotheliocyte damage and is related to rapid depression of gene expression.

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罗涛,张强,张建,李建新,谷涌泉,俞恒锡,汪忠镐.早期炎性反应对移植静脉中外源基因表达效率的影响[J].中国普通外科杂志,2006,15(5):9-349.
DOI:10.7659/j. issn.1005-6947.2006.05.009

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  • 在线发布日期: 2006-05-25