Open AccessAdenovirus viral interleukin‐10 inhibits adhesion molecule expressions induced by hypoxia/reoxygenation in cerebrovascular endothelial cells 1
Author(s) -
KANG Hui,
YANG Pengyuan,
RUI Yaocheng
Publication year - 2008
Publication title -
acta pharmacologica sinica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.514
H-Index - 90
eISSN - 1745-7254
pISSN - 1671-4083
DOI - 10.1111/j.1745-7254.2008.00718.x
Subject(s) - multiplicity of infection , cell adhesion molecule , microbiology and biotechnology , endothelial stem cell , transfection , cell adhesion , chemistry , cell culture , intercellular adhesion molecule 1 , cytokine , recombinant dna , interleukin , adhesion , monocyte , cell , biology , immunology , in vitro , biochemistry , gene , genetics , organic chemistry
Aim : To investigate the effects of recombinant adenovirus encoding viral interleukin‐10 (vIL‐10), a potent anti‐inflammatory cytokine, on adhesion molecule expressions and the adhesion rates of leukocytes to endothelial cells in cerebrovascular endothelial cells injured by hypoxia/reoxygenation (H/R). Methods : A recombinant adenovirus expressing vIL‐10 (Ad/vIL‐10 (or the green fluorescent protein (Ad/GFP) gene was constructed. A cerebrovascular endothelial cell line bEnd.3 was pretreated with a different multiplicity of infection (MOI) of Ad/vIL‐10 or Ad/GFP and then exposed to hypoxia for 9 h followed by reoxygenation for 12 h. The culture supernatants were tested for the expression of vIL‐10 and endogenous murine IL‐10 (mIL‐10) by ELISA. The effects of Ad/vIL‐10 on monocyte‐endothelial cell adhesion were represented as the adhesion rate. Subsequently, the expressions of intercellular adhesion molecule 1(ICAM‐1) and vascular cell adhesion molecule 1(VCAM‐1) in the endothelial cells after treatment with Ad/vIL‐10 and H/R were analyzed by Western blotting and real‐time PCR Results : vIL‐10 was expressed in cultured bEnd.3 after Ad/vIL‐10 transfection and was significantly increased by H/R. Ad/vIL‐10 or Ad/GFP did not affect the mIL‐10 level. H/R increased the mIL‐10 expression, but insignificantly. Monocyte‐endothelial cell adhesion induced by H/R was significantly inhibited by pretreatment with Ad/vIL‐10 (MOI: 80). ICAM‐1, and VCAM‐1 in bEnd.3 and were significantly increased after H/R, while pretreatment with Ad/vIL‐10 (MOI: 80) significantly inhibited their expressions. Ad/GFP did not markedly affect monocyte‐endothelial adhesion and the expressions of ICAM‐1 and VCAM‐1 induced by H/R. Conclusion : Ad/vIL‐10 significantly inhibits the upregulation of endothelial adhesion molecule expressions and the increase of adhesion of monocytes‐endothelial cells induced by H/R, indicating that vIL‐10 gene transfer is of far‐reaching significance in the therapy of cerebrovascular inflammatory diseases, and anti‐adhesion treatment may reduce H/R injury.