Basic Research
Copyright ©2005 Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. May 28, 2005; 11(20): 3080-3084
Published online May 28, 2005. doi: 10.3748/wjg.v11.i20.3080
Inhibition of PMA-induced endothelial cell activation and adhesion by over-expression of domain negative IκBα protein
Jian-Feng Wei, Ke Sun, Shi-Guo Xu, Hai-Yang Xie, Shu-Sen Zheng
Jian-Feng Wei, Shi-Guo Xu, Hai-Yang Xie, Shu-Sen Zheng, Key Lab of Multi-organ Transplantation of Ministry, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003, Zhejiang Province, China
Ke Sun, Department of Pathology, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003, Zhejiang Province, China
Author contributions: All authors contributed equally to the work.
Supported by the Key Lab of Multi-organ Transplantation of Ministry, College of Medicine, Zhejiang University, Hangzhou, China
Correspondence to: Shu-Sen Zheng, Key Lab of Multi-organ Transplantation of Ministry, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003, Zhejiang Province, China. zhengss@mail.hz.zj.cn
Telephone: +86-571-87236567 Fax: +86-571-87236570
Received: June 21, 2003
Revised: June 22, 2003
Accepted: October 12, 2003
Published online: May 28, 2005
Abstract

AIM: NF-κB, regulate the expression of cytokine-inducible genes involving immune and inflammatory responses, will be potential therapy approach for allograft from rejection. In this study, we use pCMV-IκBαM vector to inhibit NF-κB activation and investigate the effect of pCMV-IκBαM in inhibition of T cells adhesion to endothelial cells.

METHODS: The NF-κB activity was detected with pNF-κB reporter gene and electrophoretic mobility shift assay. Expression of cell surface molecules was detected by RT-PCR and flow cytometer. The cell-cell adhesion assay was performed to determine the effect of pCMV-IκBαM in inhibition of T cells adhesion to endothelial cells.

RESULTS: We could find that NF-κB activity is inhibited by over-expression of non-degraded IκBα protein. Expression of adhesion molecules like ICAM-1, VCAM-1, and P-selectin as well as cell-cell adhesion were inhibited significantly by transfection of the pCMV-IκBαM vector.

CONCLUSION: Our results indicate that the pCMV-IκBαM, which inhibit the activity of NF-κB through over-expression of non-degraded IκBα protein, can be used for gene therapy in diseases involving NF-κB activation abnormally like organ transplantation via inhibiting cell adhesion.

Keywords: Cytokine-inducible genes; Endothelial cells