Abstract:Objective:To investigate the effects of human arresten gene transfection on the intimal hyperplasia of venous autografts in vivo.
Methods :Autogenous vein graft model was established in male Sprague-Dawley rats. The transplanted veins were immersed in the liposome mediated recombinant eukaryotic expression plasmid pSecTag2-AT solution(group Ⅰ) or the liposome mediated control plasmid pSecTag2 solution(group Ⅱ) for 30minutes just before vascular anastomosis. Normal control group(group Ⅲ) was only treated with isovolume of liposome. Vein graft samples were harvested at 4 weeks after operation. RT-PCR was used to detect the expression of arresten mRNA in blood vessel; the surface area and thickness of the intima and media were measured by computerized planimetry under a light microscope to compare the degree of neointimal hyperplasia by the calculated ratio between intima (I) and media (M) after staining with hematoxylin-eosin and Verhoeff(elastic fibers). Immunohistochemical labeling and morphologic analysis of vein graft sections were used to identify PCNA positive cells and α-SMA positive cells; Western blot was used to detect the protein of TGF-β1.
Results:The genome of arresten-transferred tissue contained a 449bp specific fragment of arresten gene was expressed in group Ⅰ， but was not expressed in group Ⅱ or group Ⅲ; the surface area of the intima and media of group Ⅰ was less than that of group Ⅱ and group Ⅲ, and the difference were statistically significant (P<0.05), while I/M had no statistical difference (P>0.05). A less intimal thickness of groupⅠ was seen compared with group Ⅱ and group Ⅲ (P<0.01). α-SMA staining suggested that VSMC was present in the hyperplastic intimal. The number of PCNA-positive-stained cells and expression index of group Ⅰ was lower as compared with that of group Ⅱ and group Ⅲ (P<0.05). Protein level of TGF-β1 of group Ⅰ decreased obviously compared with group Ⅱ and group Ⅲ.
Conclusions:Transfection of human arresten gene can effectively inhibit the intimal hyperplasia of venous autograft. Arresten has a potential perspective for clinic application in prevention and treatment of restenosis after vascular transplantation.