Inflammatory responses involving tumor necrosis factor receptor-associated factor 6 contribute to in-stent lesion formation in a stent implantation model.

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Presentation transcript:

Inflammatory responses involving tumor necrosis factor receptor-associated factor 6 contribute to in-stent lesion formation in a stent implantation model of rabbit carotid artery Takuya Miyahara, MD, Hiroyuki Koyama, MD, Tetsuro Miyata, MD, Hiroshi Shigematsu, MD, Jun-Ichiro Inoue, MD, Tsuyoshi Takato, MD, Hirokazu Nagawa, MD Journal of Vascular Surgery Volume 43, Issue 3, Pages 592-600 (March 2006) DOI: 10.1016/j.jvs.2005.11.010 Copyright © 2006 The Society for Vascular Surgery Terms and Conditions

Fig 1 Experimental protocol. DN-TRAF6, Dominant negative tumor necrosis factor receptor-associated 6. Journal of Vascular Surgery 2006 43, 592-600DOI: (10.1016/j.jvs.2005.11.010) Copyright © 2006 The Society for Vascular Surgery Terms and Conditions

Fig 2 Photomicrographs of (A) control plasmid-treated vessel stained with anti-FLAG tag antibody, (B to E) dominant negative tumor necrosis factor receptor-associated 6 (DN-TRAF6) plasmid-treated vessel stained with anti-FLAG tag antibody, (F) control plasmid-treated vessel stained with nonspecific immunoglobulin G (IgG), (G) DN-TRAF6 plasmid-treated vessel stained with nonspecific IgG, and (H) control vessel in which phosphate-buffered saline was instilled into the luminal space followed by electroporation and stained with anti-FLAG tag antibody. Electroporation was performed at 28 days after the first injury, and the carotid artery was resected at 2 days (A, B, and F to H); 3 days (C); 7 days (D); and 14 days (E) after gene transfer. Sections were stained with anti-FLAG tag antibody or nonspecific IgG and counterstained with hematoxylin. L, Lumen; I (bar with arrows), intima; M, media. Arrowhead, internal elastic lamina. Bar = 100 μm. Journal of Vascular Surgery 2006 43, 592-600DOI: (10.1016/j.jvs.2005.11.010) Copyright © 2006 The Society for Vascular Surgery Terms and Conditions

Fig 3 Photomicrographs of (A) control plasmid-treated vessel and (B) dominant negative tumor necrosis factor receptor-associated 6 (DN-TRAF6) plasmid-treated vessel at 14 days after stent implantation. Sections were stained with hematoxylin and eosin. L, Lumen; I (bar with arrows), intima; M, media. *Strut hole; arrowhead, internal elastic lamina. Bar = 100 μm. C, Time course of intimal area. D, intimal cell number. E, intimal cell density in intima of control plasmid-treated vessel (open bars) and DN-TRAF6 plasmid-treated vessel (solid bars). Control values (hatched bars) are from rabbits 28 days after first injury without stent implantation. Values are shown as mean ± SEM. (*P < .01, **P < .05 vs control without stent implantation, #P < .01, §P < .05 control group vs DN-TRAF6 group). C, Control; d, days after stent implantation. Journal of Vascular Surgery 2006 43, 592-600DOI: (10.1016/j.jvs.2005.11.010) Copyright © 2006 The Society for Vascular Surgery Terms and Conditions

Fig 4 Photomicrographs of (A) control plasmid-treated vessel and (B) dominant negative tumor necrosis factor receptor-associated 6 (DN-TRAF6) plasmid-treated vessel at 7 days after stent implantation. Sections were stained with anti-5-bromo-2′-deoxyuridine (BrdU) antibody and counterstained with hematoxylin. BrdU-positive cells stain brown. L, Lumen; I (bar with arrows), intima; M, media. *Strut hole; arrowhead, internal elastic lamina. Bar = 100 μm. C, Time course of intimal BrdU index. D, medial BrdU index in intima of control plasmid treated-vessel (open bars) and DN-TRAF6 plasmid-treated vessel (solid bars). Control values (hatched bars) are from rabbits 28 days after first injury without stent implantation. Values are shown as mean ± SEM. (*P < .01 vs control without stent implantation, #P < .01 control group vs DN-TRAF6 group). C, Control; d, days after stent implantation. Journal of Vascular Surgery 2006 43, 592-600DOI: (10.1016/j.jvs.2005.11.010) Copyright © 2006 The Society for Vascular Surgery Terms and Conditions

Fig 5 Photomicrographs of (A) control plasmid-treated vessel and (B) dominant negative tumor necrosis factor receptor-associated 6 (DN-TRAF6) plasmid-treated vessel at 7 days after stent implantation. Sections were stained with antimacrophage antibody (RAM11) and counterstained with hematoxylin. Macrophages stain brown. L, Lumen; I (bar with arrows), intima; M, media. *Strut hole; arrowhead, internal elastic lamina. Bar = 100 μm. C, Time course of macrophage index in intima of control plasmid-treated vessel (open bars) and DN-TRAF6 plasmid-treated vessel (solid bars). Control value (hatched bar) is from rabbits 28 days after first injury without stent implantation. Values are shown as mean ± SEM. (*P < .01, **P < .05 vs control without stent implantation, #P < .01 control group vs DN-TRAF6 group). C, Control; d, days after stent implantation. Journal of Vascular Surgery 2006 43, 592-600DOI: (10.1016/j.jvs.2005.11.010) Copyright © 2006 The Society for Vascular Surgery Terms and Conditions

Fig 6 Scanning electron microscopic view of carotid lumen 3 days after stent implantation in (A) control group and (B) dominant negative tumor necrosis factor receptor-associated 6 (DN-TRAF6) group. Bar = 100 μm. Fewer leukocytes adhered to the luminal surface in the DN-TRAF6 group compared with the control group. Transmission electron microscopic view of carotid artery 14 days after stent implantation in (C) control group and (D) DN-TRAF6 group. Bar = 1 μm. Proteoglycan accumulation was suppressed in the DN-TRAF6 group compared with the control group. Samples were stained with 0.2% Cupromeronic blue with 0.3 mol/L magnesium chloride. Arrowhead indicates proteoglycans. C, Collagen. Journal of Vascular Surgery 2006 43, 592-600DOI: (10.1016/j.jvs.2005.11.010) Copyright © 2006 The Society for Vascular Surgery Terms and Conditions

Fig 7 Plasminogen activator zymograms developed for 24 hours show time course of PA activity in (A) intima and (B) media. Plasminogen activator activity at 54 kDa was suppressed in the dominant negative tumor necrosis factor receptor-associated 6 (DN-TRAF6) group compared with that in the control group in both the intima and media. N, Normal carotid; d, days after stent implantation. Journal of Vascular Surgery 2006 43, 592-600DOI: (10.1016/j.jvs.2005.11.010) Copyright © 2006 The Society for Vascular Surgery Terms and Conditions

Fig 8 Gelatin zymograms show time course of gelatinolytic activity in the (A) intima and (B) media. Matrix metalloproteinase activity at 95 kDa was suppressed in the dominant negative tumor necrosis factor receptor-associated 6 (DN-TRAF6) group compared with that in the control group in both the intima and media. N, Normal carotid; d, days after stent implantation. Journal of Vascular Surgery 2006 43, 592-600DOI: (10.1016/j.jvs.2005.11.010) Copyright © 2006 The Society for Vascular Surgery Terms and Conditions