Zhang Jing-hong, Sun Cheng-lin, Duan Zhi-quan, Xin Shi-jie
Department of Vascular Surgery, First Hospital Affiliated to China Medical University, Shenyang 110001, China.
Zhonghua Yi Xue Za Zhi. 2007 May 8;87(17):1207-10.
To assess the effects of transfection of adenovirus on the tunica intima hyperplasia of vein autografts.
An external branch of the internal jugular vein, 5 mm in length, of a Wistar rat was cut and transplanted to its own common carotid artery by end to end bypass so as to establish a model. Ninety Wistar rats underwent transplantation of vein autografts to their own arteries and then were randomly divided into 3 equal groups: experiment group, undergoing transplantation and adenovirus transfection; experimental control group, undergoing transplantation only; and normal control group. Three, 10, and 30 days later, samples of vessel were obtained. The expression of green fluorescence protein (GFP) was observed so as to measure the transfection rate of adenovirus. Routine HE staining and Verhoeff Van Gieson staining were made to measure the thickness of the vessels with computer-assisted image analyzer. Immunohistochemistry was used to detect the proliferating cell nuclear antigen (PCNA) RT-PCR and Western blotting were used to detect the mRNA and protein expression of intracellular adhesion molecule-1 (ICAM-1) and vascular adhesion molecule-1 (VCAM-1).
Three day postoperatively, the expression of GFP reached the peak, (26.4 +/- 3.6)%; decreased to (14.5 +/- 2.1)% 10 day postoperatively; and became very low 30 day postoperatively, at the level of (0.81 +/- 0.2)%. The hyperplasia of venous tunica intima of the control group was obvious 10 day postoperatively, and became less obvious 30 day postoperatively. Proliferation of vascular smooth muscle cells (VSMCs) and sedimentation of extra-cellular matrix in the experimental group were very obvious, compared with the other 2 groups (both P < 0.05). PCNA was very rare in normal vein walls, showing that the VSMCs were in the static phase. PCNA presentation increased obviously in the control group, showing a high proliferation rate of VSMCs. The positive rate of PCNA was associated with the thickness of the tunica intima. mRNA expression and protein expression of ICAM-1 and VCAM-1 could be detected 3 days after the transplantation, peaked 10 days after the transplantation, and remained high 30 days after in the normal group, however, the corresponding expression levels of the normal vein group were all very low (all P < 0.05). The corresponding levels of the experimental group were all higher than those of the experimental control group, however, not significantly different.
Transfection of adenovirus into the wall of transplanted vein causes inflammation and hyperplasia of tunica intima. However, such affects only a short time.
评估腺病毒转染对自体静脉移植物内膜增生的影响。
取Wistar大鼠颈内静脉一长5mm的外分支,切断后通过端端吻合移植至其自身颈总动脉,建立模型。90只Wistar大鼠行自体静脉移植物移植至自身动脉,然后随机分为3组,每组30只:实验组,行移植及腺病毒转染;实验对照组,仅行移植;正常对照组。术后3天、10天和30天取材。观察绿色荧光蛋白(GFP)表达以检测腺病毒转染率。行常规HE染色及Verhoeff Van Gieson染色,用计算机辅助图像分析仪测量血管厚度。采用免疫组化检测增殖细胞核抗原(PCNA),采用RT-PCR及Western印迹法检测细胞间黏附分子-1(ICAM-1)和血管细胞黏附分子-1(VCAM-1)的mRNA及蛋白表达。
术后3天,GFP表达达峰值,为(26.4±3.6)%;术后10天降至(14.5±2.1)%;术后30天极低,为(0.81±0.2)%。实验对照组术后10天静脉内膜增生明显,术后30天不明显。与其他两组相比,实验组血管平滑肌细胞(VSMC)增殖及细胞外基质沉积均非常明显(均P<0.05)。正常静脉壁PCNA极少,表明VSMC处于静止期。实验对照组PCNA表达明显增加,表明VSMC增殖率高。PCNA阳性率与内膜厚度相关。移植后3天可检测到ICAM-1和VCAM-1的mRNA及蛋白表达,移植后10天达峰值,正常组术后30天仍维持在较高水平,而正常静脉组相应表达水平均很低(均P<0.05)。实验组相应水平均高于实验对照组,但差异无统计学意义。
腺病毒转染移植静脉壁可引起内膜炎症及增生。但这种影响仅持续较短时间。