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...
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doaj-36ca67c9b84645168dce3895bbbef0612020-11-25T01:05:34ZengBMCJournal 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 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
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 |
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 |
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 |
title |
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_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_sort |
preventing intimal thickening of vein grafts in vein artery bypass using stat-3 sirna |
publisher |
BMC |
series |
Journal of Translational Medicine |
issn |
1479-5876 |
publishDate |
2012-01-01 |
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> |
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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