[人组织因子途径抑制物基因转染对静脉移植物新生内膜形成的影响]
[Influence of human tissue factor pathway inhibitor gene transfection on neointima formation in vein grafts].
作者信息
Li Quan, Zhang Kailun, Jiang Xionggang, Sun Tucheng, Wu Long, Zhou Cheng, Chen Shu
机构信息
Department of Cardiovascular Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan Hubei, 430022, P. R. China.
出版信息
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2008 Mar;22(3):354-8.
OBJECTIVE
To reduce restenosis in vein grafts after coronary artery bypass grafting, to investigate the effect of human tissue factor pathway inhibitor (TFPI) gene delivery on neointima formation.
METHODS
The eukaryotic expressed plasmid vector pCMV-(Kozak) TFPI was constructed. Forty-eight Japanese white rabbits were randomly divided into 3 groups with 16 rabbits in each group: TFPI group, empty plasmid control group and empty control group. Animal model of common carotid artery bypass grafting was constructed. Before anastomosis, vein endotheliocytes were transfected with cationic liposome containing the plasmid pCMV-(Kozak) TFPI (400 microg) by pressurizing infusion (30 min) in TFPI group. In empty plasmid control group, vector pCMV-(Kozak) TFPI was replaced by empty plasmid pCMV (400 microg). In empty control group, those endotheliocytes were not interfered. After operation, vein grafts were harvested at 3 days for immunohistochemical, RT-PCR and Western-blot analyses of exogenous gene expression and at 30 days for histopathology measurement of intimal areas, media areas and calculation of intimal/media areas ratio. Luminal diameter and vessel wall thickness were also measured by vessel Doppler ultrasonography and cellular category of neointima was analyzed by transmission electron microscope at 30 days after operation.
RESULTS
Human TFPI mRNA and protein were detected in TFPI group. The mean luminal diameter of the TFPI group, empty plasmid control group and empty control group was (2.68 +/- 0.32) mm, (2.41 +/- 0.23) mm and (2.38 +/- 0.21) mm respectively. There were statistically significant differences between TFPI group and control groups (P < 0.05). The vessel wall thickness of the TFPI group, empty plasmid control group and empty control group was (1.09 +/- 0.11) mm, (1.28 +/- 0.16) mm and (1.34 +/- 0.14) mm respectively. There were statistically significant differences between TFPI group and other control groups (P < 0.01). The mean intimal areas, the ratio of the intimal/media areas of the TFPI group were (0.62 +/- 0.05) mm2 and 0.51 +/- 0.08 respectively,whichwere reduced compared withthose of the two control groups (P < 0.05). The mean media areas had no significant differences among three groups (P > 0.05). Through transmission electron microscope analyses,no smooth muscle cells were seen in neointima of TFPI group in many visual fields,but smooth muscle cells were found in neointima of two control groups.
CONCLUSION
Human TFPI gene transfection reduced intimal thickness in vein grafts.
目的
为减少冠状动脉旁路移植术后静脉移植物再狭窄，研究人组织因子途径抑制物（TFPI）基因转染对新生内膜形成的影响。
方法
构建真核表达质粒载体pCMV-(Kozak)TFPI。48只日本大耳白兔随机分为3组，每组16只：TFPI组、空质粒对照组和空白对照组。建立颈总动脉旁路移植动物模型。在吻合前，TFPI组通过加压输注（30分钟）用含质粒pCMV-(Kozak)TFPI（400μg）的阳离子脂质体转染静脉内皮细胞。空质粒对照组，用空质粒pCMV（400μg）代替载体pCMV-(Kozak)TFPI。空白对照组，不干预这些内皮细胞。术后3天取静脉移植物进行免疫组织化学、RT-PCR和Western-blot分析外源基因表达；术后30天取静脉移植物进行组织病理学测量内膜面积、中膜面积并计算内膜/中膜面积比值。术后30天用血管多普勒超声测量管腔直径和血管壁厚度，并用透射电镜分析新生内膜的细胞种类。
结果
TFPI组检测到人TFPI mRNA和蛋白。TFPI组、空质粒对照组和空白对照组的平均管腔直径分别为(2.68±0.32)mm、(2.41±0.23)mm和(2.38±0.21)mm。TFPI组与对照组之间有统计学差异（P<0.05）。TFPI组、空质粒对照组和空白对照组的血管壁厚度分别为(1.09±0.11)mm、(1.28±0.16)mm和(1.34±0.14)mm。TFPI组与其他对照组之间有统计学差异（P<0.01）。TFPI组的平均内膜面积、内膜/中膜面积比值分别为(0.62±0.05)mm²和0.51±0.08，与两个对照组相比降低（P<0.05）。三组间平均中膜面积无显著差异（P>0.05）。通过透射电镜分析，TFPI组许多视野的新生内膜未见平滑肌细胞，而两个对照组的新生内膜可见平滑肌细胞。
结论
人TFPI基因转染可减少静脉移植物的内膜厚度。
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