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. Author manuscript; available in PMC: 2017 Mar 17.
Published in final edited form as: Circulation. 2013 Sep 10;128(11 Suppl 1):S50–S58. doi: 10.1161/CIRCULATIONAHA.112.000249

Figure 1. Endothelial overexpression of human GTP cyclohydrolase (GCH) reduces neointimal hyperplasia and enhances repopulation and survival of endothelial cell (EC) in vein grafts.

Figure 1

A, When vena cava was grafted into recipients of the same genotype, vein graft wall area at 28 days after surgery was significantly lower in GCH/apolipoprotein E (apoE)-knockout (KO; n=5) mice compared with apoE-KO mice (n=8). When vena cava of apoE-KO mice was grafted into GCH/apoE-KO recipients (n=6), the mean vessel wall area was similar to that seen when GCH/apoE-KO vena cava was grafted into GCH/apoE-KO recipients. When GCH/apoE-KO donor vena cava was grafted into apoE-KO recipients (n=8), there was a less marked, but still significant, reduction in lesion area compared with apoE-KO vena cava grafted into apoE-KO. Black bar represents 100 μm. One-way ANOVA overall P<0.0001. Bonferroni multiple comparison test *P<0.01 vs apoE-KO vein grafted into apoE-KO recipient and †P<0.05 vs GCH/apoE-KO vein grafted into apoE-KO. B, Tetrahydrobiopterin (BH4) levels in harvested vein grafts were significantly higher in GCH/apoE-KO compared with apoE-KO. When apoE-KO veins were grafted into GCH/apoE-KO mice, graft BH4 levels reached levels similar to the GCH/apoE-KO vein grafts. When GCH/apoE-KO veins were grafted into apoE-KO mice, BH4 levels were significantly higher than apoE-KO vein grafts grafted into matched recipients (n=5). One-way ANOVA overall P<0.0001. Bonferroni multiple comparison test *P<0.05 vs apoE-KO. †P<0.05 vs GCH/apoE-KO vein grafted into apoE-KO recipients. C, Vein graft mRNA levels of total GCH (transgenic+endogenous) were significantly higher in GCH/apoE-KO mice compared with apoE-KO. When apoE-KO veins were grafted into GCH/apoE-KO mice, mRNA levels were similar to the GCH/apoE-KO vein grafts. One-way ANOVA overall P<0.0001. Bonferroni multiple comparison test *P<0.001 vs apoE-KO. †P<0.001 vs GCH/apoE-KO vein grafted into apoE-KO recipients (n=4). D, Vein graft endogenous mouse GCH mRNA levels were not different in GCH/apoE-KO mice compared with apoE-KO, regardless of the source of the vein (n=4). E, When apoE-KO veins were grafted into apoE-KO/LacZ recipients, the presence of β-galactosidase (β-Gal)–positive staining (blue) on the surface of vein grafts at 28 days indicated recipient-derived EC repopulation (n=5). When GCH/apoE-KO veins were grafted into GCH/apoE-KO/LacZ recipients (n=6), EC coverage was increased compared with grafting apoE-KO veins into apoE-KO/LacZ recipients, indicating enhanced recipient-derived EC repopulation in GCH animals. Investigating EC survival, no β-Gal staining in apoE-KO/LacZ veins grafted into apoE-KO recipients could be detected. However, when GCH/apoE-KO/LacZ veins were grafted into GCH/apoE-KO recipients (n=5), a small proportion of β-Gal–positive cells remained evident on the luminal surface of the vein graft at 28 days, indicating enhanced EC survival from GCH transgenic donors. No β-Gal–positive cells were visualized in apoE-KO/LacZ veins grafted into apoE-KO recipients (n=5). Arrowhead denotes β-Gal–positive cell. Unpaired t test *P=0.02 vs apoE-KO/LacZ recipient. †P=0.02 vs apoE-KO recipient.