INHIBITION OF CYTOKINE EXPRESSION IN HUMAN MESANGIAL CELLS WITH
NF-kB ANTISENSE
OLIGODEOXYNUCLEOTIDE
Ding G-X,Chen RH
Nanjing Medical University, Nanjing, China
Objective: The blocking effects in NF-kB antisense oilgodeoxynucleotide
(AS-ODN) on the pro-inflammatory cytokines,
chemokines, and pro-fibrogentic cytokine expression of human mesangial
cells (HMC) was investigated to probe the potential applicability of the
ASODN drug in renal inflammatory disorders.
Methods: AS-ODN, sense (S)-ODN, and
mismatch (Mis)-ODN were manually synthesized and completely
phosphorothioate-modified. Cationic lipid-mediated ODN were transferred
into HMC. TNF-a, IL-1a, IL-1b, MCP-1, IL-8, and TGF-b1 mRNA
expression was determined by RPA. The levels of p65 mRNA and protein expression were measured by
RT-PCR and Western Blot, respectively.
Results: HMC which was incubated in medium alone did not express IL-1a and MCP-1 mRNA, and constitutive mRNA expression in
unstimulated cells was found for TNF-a, IL-1b,
IL-8, and TGF-b1. P65 ASODN
inhibited TNF-a, IL-1a, IL-1b,
MCP-1 and IL-8 mRNA expression in LPS-stimulated HMC in a dose-dependent
manner, as compared with that treated with sense p65 ODN and mismatch ODN.
Three ODN has no effect on the TGF-b1 mRNA expression of HMC. p65 ASON
inhibited the expression of p65 mRNA and protein in dose-dependent fasion.
Conclusion: p65 ASODN can inhibit proinflammatory
cytokines, chemokines, and profibrogenic cytokine synthesis of the cells,
which suggests that p65 ASODN has important regulatory effects on the
inflammatory response of HMC and might be applied in treatment of renal
inflammatory disorders.