在用α-黑素细胞刺激素载体转染的人肺上皮细胞中，核因子κB的激活受到抑制。

NF-kappaB activation is inhibited in human pulmonary epithelial cells transfected with alpha-melanocyte-stimulating hormone vector.

作者信息

Ichiyama T, Okada K, Campbell I L, Furukawa S, Lipton J M

机构信息

Department of Physiology, University of Texas Southwestern Medical Center at Dallas, Dallas, TX 75235-9040, USA.

出版信息

Peptides. 2000 Oct;21(10):1473-7. doi: 10.1016/s0196-9781(00)00300-4.

DOI:10.1016/s0196-9781(00)00300-4

PMID:11068093

Abstract

alpha-Melanocyte-stimulating hormone (alpha-MSH) modulates inflammation. We investigated the influence of alpha-MSH on NF-kappaB activation in human pulmonary epithelial cells (A549) using a plasmid vector encoding alpha-MSH (pCMV-ssMSH). Electrophoretic mobility shift assays demonstrated that NF-kappaB activation induced by lipopolysaccharide was inhibited in A549 cells transfected with pCMV-ssMSH. Western blot analysis revealed that this inhibition was linked to preservation of expression of IkappaBalpha protein. Chloramphenicol acetyltransferase assay indicated that NF-kappaB-dependent reporter gene expression was suppressed in A549 cells transfected with pCMV-ssMSH. The findings indicate that anti-inflammatory actions are exerted via modulation of NF-kappaB activation by preservation of IkappaBalpha protein in human pulmonary epithelial cells transfected with alpha-MSH vector. We showed a possibility of gene therapy for chronic inflammatory lung diseases.

摘要

α-黑素细胞刺激素（α-MSH）可调节炎症。我们使用编码α-MSH的质粒载体（pCMV-ssMSH）研究了α-MSH对人肺上皮细胞（A549）中NF-κB激活的影响。电泳迁移率变动分析表明，在用pCMV-ssMSH转染的A549细胞中，脂多糖诱导的NF-κB激活受到抑制。蛋白质免疫印迹分析显示，这种抑制作用与IκBα蛋白表达的维持有关。氯霉素乙酰转移酶分析表明，在用pCMV-ssMSH转染的A549细胞中，NF-κB依赖性报告基因的表达受到抑制。这些发现表明，在用α-MSH载体转染的人肺上皮细胞中，通过维持IκBα蛋白来调节NF-κB激活，从而发挥抗炎作用。我们展示了慢性炎症性肺病基因治疗的可能性。