Preventing intimal thickening of vein grafts in vein artery bypass using STAT-3 siRNA
<p>Abstract</p> <p>Background</p> <p>Proliferation and migration of vascular smooth muscle cells (VSMCs) play a key role in neointimal formation which leads to restenosis of vein graft in venous bypass. STAT-3 is a transcription factor associated with cell proliferation...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2012-01-01
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| Series: | Journal of Translational Medicine |
| Subjects: | |
| Online Access: | http://www.translational-medicine.com/content/10/1/2 |
| _version_ | 1818826786493956096 |
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| author | Sun Jiangbin Zheng Jinhua Ling Kaitelynne H Zhao Keyan Xie Zhongshang Li Bo Wang Tiance Zhu Zhicheng Patel Amit N Min Weiping Liu Kexiang Zheng Xiufen |
| author_facet | Sun Jiangbin Zheng Jinhua Ling Kaitelynne H Zhao Keyan Xie Zhongshang Li Bo Wang Tiance Zhu Zhicheng Patel Amit N Min Weiping Liu Kexiang Zheng Xiufen |
| author_sort | Sun Jiangbin |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>Proliferation and migration of vascular smooth muscle cells (VSMCs) play a key role in neointimal formation which leads to restenosis of vein graft in venous bypass. STAT-3 is a transcription factor associated with cell proliferation. We hypothesized that silencing of STAT-3 by siRNA will inhibit proliferation of VSMCs and attenuate intimal thickening.</p> <p>Methods</p> <p>Rat VSMCs were isolated and cultured in vitro by applying tissue piece inoculation methods. VSMCs were transfected with STAT 3 siRNA using lipofectamine 2000. In vitro proliferation of VSMC was quantified by the MTT assay, while in vivo assessment was performed in a venous transplantation model. In vivo delivery of STAT-3 siRNA plasmid or scramble plasmid was performed by admixing with liposomes 2000 and transfected into the vein graft by bioprotein gel applied onto the adventitia. Rat jugular vein-carotid artery bypass was performed. On day 3 and7 after grafting, the vein grafts were extracted, and analyzed morphologically by haematoxylin eosin (H&E), and assessed by immunohistochemistry for expression of Ki-67 and proliferating cell nuclear antigen (PCNA). Western-blot and reverse transcriptase polymerase chain reaction (RT-PCR) were used to detect the protein and mRNA expression in vivo and in vitro. Cell apoptosis in vein grafts was detected by TUNEL assay.</p> <p>Results</p> <p>MTT assay shows that the proliferation of VSMCs in the STAT-3 siRNA treated group was inhibited. On day 7 after operation, a reduced number of Ki-67 and PCNA positive cells were observed in the neointima of the vein graft in the STAT-3 siRNA treated group as compared to the scramble control. The PCNA index in the control group (31.3 ± 4.7) was higher than that in the STAT-3 siRNA treated group (23.3 ± 2.8) (P < 0.05) on 7d. The neointima in the experimental group(0.45 ± 0.04 μm) was thinner than that in the control group(0.86 ± 0.05 μm) (P < 0.05).Compared with the control group, the protein and mRNA levels in the experimental group in vivo and in vitro decreased significantly. Down regulation of STAT-3 with siRNA resulted in a reduced expression of Bcl-2 and cyclin D1. However, apoptotic cells were not obviously found in all grafts on day 3 and 7 post surgery.</p> <p>Conclusions</p> <p>The STAT-3 siRNA can inhibit the proliferation of VSMCs in vivo and in vitro and attenuate neointimal formation.</p> |
| first_indexed | 2024-12-19T00:33:12Z |
| format | Article |
| id | doaj.art-36ca67c9b84645168dce3895bbbef061 |
| institution | Directory Open Access Journal |
| issn | 1479-5876 |
| language | English |
| last_indexed | 2024-12-19T00:33:12Z |
| publishDate | 2012-01-01 |
| publisher | BMC |
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| series | Journal of Translational Medicine |
| spelling | doaj.art-36ca67c9b84645168dce3895bbbef0612022-12-21T20:45:00ZengBMCJournal of Translational Medicine1479-58762012-01-01101210.1186/1479-5876-10-2Preventing intimal thickening of vein grafts in vein artery bypass using STAT-3 siRNASun JiangbinZheng JinhuaLing Kaitelynne HZhao KeyanXie ZhongshangLi BoWang TianceZhu ZhichengPatel Amit NMin WeipingLiu KexiangZheng Xiufen<p>Abstract</p> <p>Background</p> <p>Proliferation and migration of vascular smooth muscle cells (VSMCs) play a key role in neointimal formation which leads to restenosis of vein graft in venous bypass. STAT-3 is a transcription factor associated with cell proliferation. We hypothesized that silencing of STAT-3 by siRNA will inhibit proliferation of VSMCs and attenuate intimal thickening.</p> <p>Methods</p> <p>Rat VSMCs were isolated and cultured in vitro by applying tissue piece inoculation methods. VSMCs were transfected with STAT 3 siRNA using lipofectamine 2000. In vitro proliferation of VSMC was quantified by the MTT assay, while in vivo assessment was performed in a venous transplantation model. In vivo delivery of STAT-3 siRNA plasmid or scramble plasmid was performed by admixing with liposomes 2000 and transfected into the vein graft by bioprotein gel applied onto the adventitia. Rat jugular vein-carotid artery bypass was performed. On day 3 and7 after grafting, the vein grafts were extracted, and analyzed morphologically by haematoxylin eosin (H&E), and assessed by immunohistochemistry for expression of Ki-67 and proliferating cell nuclear antigen (PCNA). Western-blot and reverse transcriptase polymerase chain reaction (RT-PCR) were used to detect the protein and mRNA expression in vivo and in vitro. Cell apoptosis in vein grafts was detected by TUNEL assay.</p> <p>Results</p> <p>MTT assay shows that the proliferation of VSMCs in the STAT-3 siRNA treated group was inhibited. On day 7 after operation, a reduced number of Ki-67 and PCNA positive cells were observed in the neointima of the vein graft in the STAT-3 siRNA treated group as compared to the scramble control. The PCNA index in the control group (31.3 ± 4.7) was higher than that in the STAT-3 siRNA treated group (23.3 ± 2.8) (P < 0.05) on 7d. The neointima in the experimental group(0.45 ± 0.04 μm) was thinner than that in the control group(0.86 ± 0.05 μm) (P < 0.05).Compared with the control group, the protein and mRNA levels in the experimental group in vivo and in vitro decreased significantly. Down regulation of STAT-3 with siRNA resulted in a reduced expression of Bcl-2 and cyclin D1. However, apoptotic cells were not obviously found in all grafts on day 3 and 7 post surgery.</p> <p>Conclusions</p> <p>The STAT-3 siRNA can inhibit the proliferation of VSMCs in vivo and in vitro and attenuate neointimal formation.</p>http://www.translational-medicine.com/content/10/1/2STAT-3, siRNAvascular smooth muscle cells (VSMCs)intimal thickening |
| spellingShingle | Sun Jiangbin Zheng Jinhua Ling Kaitelynne H Zhao Keyan Xie Zhongshang Li Bo Wang Tiance Zhu Zhicheng Patel Amit N Min Weiping Liu Kexiang Zheng Xiufen Preventing intimal thickening of vein grafts in vein artery bypass using STAT-3 siRNA Journal of Translational Medicine STAT-3, siRNA vascular smooth muscle cells (VSMCs) intimal thickening |
| title | Preventing intimal thickening of vein grafts in vein artery bypass using STAT-3 siRNA |
| title_full | Preventing intimal thickening of vein grafts in vein artery bypass using STAT-3 siRNA |
| title_fullStr | Preventing intimal thickening of vein grafts in vein artery bypass using STAT-3 siRNA |
| title_full_unstemmed | Preventing intimal thickening of vein grafts in vein artery bypass using STAT-3 siRNA |
| title_short | Preventing intimal thickening of vein grafts in vein artery bypass using STAT-3 siRNA |
| title_sort | preventing intimal thickening of vein grafts in vein artery bypass using stat 3 sirna |
| topic | STAT-3, siRNA vascular smooth muscle cells (VSMCs) intimal thickening |
| url | http://www.translational-medicine.com/content/10/1/2 |
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