| Objective: To investigate the effect to c-myc antisense oligonucleotides (ASODN) combinedRe targeting c-myc proto oncogene on the neointimal thickening in rabbit common iliac arteria injured by the porous balloon catheter to explore the therapeutic potential of c-myc ASODNs for prevention of restenosis.Methods: After the radian therapy with the 2.5mm balloon catheter filled with 188Re, c-myc ASODNs was applied to the adventitia of injured common iliac arteria in rabbit introduced by pluronic gel releasing system and lipofectin delivery system. Three weeks after intervention, the animals were killed and defined segments of arteries were sectioned and the histological sections were stained with HE staining and α -actin immunohistochemistry staining. The cross-section of the artery was examined microscopically and the image was digitalized by image analysis system. The intima/media(I/M) area and positive α —actin ratios were calculated and analyzed statistically among groups.Results:1 .Morphological change: 3 weeks after balloon injuries in rabbits, the cave of the iliac artery was extremely narrow and neointima was formed and the media layer (VSM layer) was proliferated.2. Effects of ASODN to the changes of cave, neointima and media areas: In the ASODN group, the areas of the vessel caves from operation side were larger than those in control group(0.91 ± 0.23 mm2 vs 0.72 ± 0.19 mm2, P<0.01), the intima areas were declined (0.20± 0.12 mm2 vs 0.62 ± 0.18 mm, P<0.01) , and the ratios of intima to media area were declined too (0.34 ± 0.05 vs 1.02 ± 0.15, P<0.01)。3. Effects of 188Re radian to the changes of cave, neointima and media areas: (1) In the 8Gy dosage 188Re group, the areas of the vessel caves from operation side were no larger than those in control group (0.65 ± 0.18 mm2vs 0.72 ± 0.19 mm2, P>0.05) , the intima areas were not declined (0.52 ± 0.17 mm2 vs 0.62 ± 0.18 mm2, P>0.05), and the ratios of intima to media area were not declined too(0.92±0.09 vs 1.02±0.15, P>0.05) 。(2) In the 15Gy dosage 188Re group, the areas of the vessel caves from operation side were larger than those in control group (0.90 ±0.22 mm2 vs 0.72±0.19 mm2, P<0.01), the intima areas were declined (0.25 ± 0.13 mm2 vs0.62 ±0.18 mm2, PO.01) , and the ratios of intima to media area were declined too (0.42± 0.09vj1.02±0.15,P<0.01).4. Effects of ASODN combined 188Re radian to the changes of cave, neointima and media areas: (1) In the ASODN combined 15Gy dosage 188Re group, the areas of the vessel caves from operation side were larger than those in control group (0.95 ±0.21 mm2 vs 0.72 ±0.19 mm2, ?<0.01), the intima areas were declined(0.04±0.015 mm2 vs 0.62±0.18 mm2, PO.01), and the ratios of intima to media area were declined too (0.08 + 0.018 vs 1.02 ±0.15, PO.01 )? The areas of the vessel caves from operation side were no larger than those in ASODN group(0.95 + 0.21 mm2 vs 0.91 +0.23 mm2, P>0.05), the intima areas were declined (0.04+0.015 mm2 vs 0.20 ±0.12 mm , .P<0.01), and the ratios of intima to media area were declined too(0.08 + 0.018 vs 0.34±0.05, PO.01). The areas of the vessel caves from operation side were no larger than those in 15Gy dosage I88Re group (0.95±0.21 mm2 vs 0.90±0.22 mm2, P>0.05), the intima areas were declined (0.04±0.015 mm2 vs 0.25±0.13 mm2, PO.01), and the ratios of intima to media area were declined too (0.08 ±0.018 vs 0.40 + 0.09, P<0.0\). (2) In the ASODN combined 8Gy dosage 188Re group, the areas of the vessel caves from operation side were larger than those in control group(0.91 ±0.22 mm2 vs 0.72±0.19 mm2, P<0.01), the intima areas were declined (0.20 ±0.14 mm2vs 0.62 ±0.18 mm2, PO.01), and the ratios of intima to media area were declined too (0.32±0.05 vs 1.02+0.15, PO.01). The areas of the vessel caves from operation side were no larger than those in ASODN group (0.91 ±0.22 mm2 vs 0.91 ±0.23 mm2, P>0.05), the intima areas were not declined (0.20 + 0.14 mm2 vs 0.20 ± 0.12 mm2, P>0.05), and the ratios of intima to media area were not declined too( 0.32 ±0.05 vs 0.34 ±0.05, P>0.05). The areas of the vessel caves from operation side were larger than those in 8Gy dosage 188Re group (0.91 ±0.22 mm2 vs 0.65 ±0.18 mm2, PO.01), the intima areas were declined (0.20 ±0.14 mm2 vs 0.52 ±0.17 mm2, P<0.01), and the ratios of intima to media area were declined too (0.32+0.05 vs 0.92 + 0.09, PO.01).5. Effects of ASODN to the migration of the VSMC: There were a —actin positive staining cells in neointima after balloon injuries indicating that there were VSMCs migration to form meointima. In ASODN group, compared with the control group, the a —actin positive staining cells rate were decreased, 0.35 ±0.10% vs 0.73 ±0.15%, P<0.0\.6. Effects of 188Re radian to the migration of the VSMC: In the 15Gy dosage 188Re radiangroup, compared with the control group, the a — actin positive staining cells rate were decreased, 0.38±0.14% vs 0.73+0.15%, P<0.01; In the 15Gy dosage 188Re radian group, compared with the control group, the a —actin positive staining cells rate were not decreased, 0.70 ± 0.15% vs 0.73 ±0.15%, P>0.05?7. Effects of ASODN combined 188Re radian to the migration of the VSMC:(1) In ASODN combined 15Gy dosage I88Re radian group, compared with the control group, the a —actin positive staining cells rate were decreased, 0.18 + 0.07% vs 0.73+0.15%, PO.01; compared with the ASODN group, the a —actin positive staining cells rate were decreased, 0.18 + 0.07% vs 0.35 + 0.10%, PO.01; compared with the 15Gy dosage 188Re radian group, the a —actin positive staining cells rate were decreased, 0.18+0.07% vs 0.38+0.14%, ?<0.01o (2) In ASODN combined 8Gy dosage 188Re radian group, compared with the control group, the a — actin positive staining cells rate were decreased, 0.36±0.11% vs 0.73±0.15%, PO.01; compared with the ASODN group, the a —actin positive staining cells rate were not decreased, 0.36+0.11% vs 0.35 + 0.10%, P>0.05; compared with the 15Gy dosage 188Re radian group, the a —actin positive staining cells rate were decreased, 0.36 + 0.11% vs 0.70 + 0.15%, P<0.01 =8. In ASODN combined 15Gy dosage 188Re radian group, compared with the ASODN combined 15Gy dosage 188Re radian group, the a —actin positive staining cells rate were decreased, 0.18 + 0.07% vs 0.36+0.11%, PO.0KConclusion:1. The effects of the oligonucleotides therapy to the vessel caves and neointima depend on the sequence: The c-myc ASODN (300 u g) can relieve the vessel stenosis through increasing cave areas and decling neointima area, while the sense oligonucleotides (300 u g) can't.2. The effects of the I88Re radian to the vessel caves and neointima depend on the dosage. The 15Gy dosage can relieve the vessel stenosis through increasing cave areas and decling neointima area, while the 8Gy dosage can't.3. The c-myc ASODNs (300 u g) combined 188Re radian to the vessel caves and neointima depend on the 188Re dosage: The c-myc ASODNs combined 15Gy dosage 188Re significantly relieve the vessel stenosis through increasing cave areas and decling neointima area, the effects are better than any one of them. The c-myc ASODNs combined 8Gy dosage 188Re significantlyrelieve the vessel stenosis through increasing cave areas and decling neointima area, but there are not significant differenence with ASODNs group. |
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