Tuberc Respir Dis.  2009 Aug;67(2):95-104.

The Effects of Nuclear Factor-kappa B Decoy Oligodeoxynucleotide on Lipopolysaccharide-Induced Direct Acute Lung Injury

• Kim JH
• Yoon DW
• Jung KH
• Kim HO
• Ha ES
• Lee KJ
• Hur GY
• Lee SY
• Lee SY
• Shin C
• Shim JJ
• In KH
• Yoo SH
• Kang KH

Abstract

BACKGROUND
The pathophysiologic mechanisms of early acute lung injury (ALI) differ according to the type of primary insult. It is important to differentiate between direct and indirect pathophysiologic pathways, and this may influence the approach to treatment strategies. NF-kappa B decoy oligodeoxynucleotide (ODN) is a useful tool for the blockade of the expression of NF-kappa B-dependent proinflammatory mediators and has been reported to be effective in indirect ALI. The purpose of this study was to investigate the effect of NF-kappa B decoy ODN in the lipopolysaccharide (LPS)-induced direct ALI model. METHODS: Five-week-old specific pathogen-free male BALB/c mice were used for the experiment. In the preliminary studies, tumor necrosis factor (TNF)-alpha, interleukine (IL)-6 and NF-kappa B activity peaked at 6 hours after LPS administration. Myeloperoxidase (MPO) activity and ALI score were highest at 36 and 48 hours, respectively. Therefore, it was decided to measure each parameter at the time of its highest level. The study mice were randomly divided into three experimental groups: (1) control group which was administered 50 microliter of saline and treated with intratracheal administration of 200 microliter DW containing only hemagglutinating virus of Japan (HVJ) vector (n=24); (2) LPS group in which LPS-induced ALI mice were treated with intratracheal administration of 200 microliter DW containing only HVJ vector (n=24); (3) LPS+ODN group in which LPS-induced ALI mice were treated with intratracheal administration of 200 microliter DW containing 160 microgram of NF-kappa B decoy ODN and HVJ vector (n=24). Each group was subdivided into four experimental subgroups: (1) tissue subgroup for histopathological examination for ALI at 48 hours (n=6); (2) 6-hour bronchoalveolar lavage (BAL) subgroup for measurement of TNF-alpha and IL-6 in BAL fluid (BALF) (n=6); (3) 36-hour BAL subgroup for MPO activity assays in BALF (n=6); and (4) tissue homogenate subgroup for measurement of NF-kappa B activity in lung tissue homogenates at 6 hours (n=6). RESULTS: NF-kappa B decoy ODN treatment significantly decreased NF-kappa B activity in lung tissues. However, it failed to improve the parameters of LPS-induced direct ALI, including the concentrations of tumor necrosis factor-alpha and interleukin-6 in BALF, myeloperoxidase activity in BALF and histopathologic changes measured by the ALI score. CONCLUSION: NF-kappa B decoy ODN, which has been proven to be effective in indirect models, had no effect in the direct ALI model.

Keyword

Acute lung injury; Lipopolysaccharides; Inflammation; Nuclear factor kappa B; Oligodeoxynucleotides

MeSH Terms

Acute Lung Injury
Animals
Bronchoalveolar Lavage
Humans
Inflammation
Interleukin-6
Interleukins
Lipopolysaccharides
Lung
Male
Mice
NF-kappa B
Oligodeoxyribonucleotides
Peroxidase
Sendai virus
Tumor Necrosis Factor-alpha
Interleukin-6
Interleukins
Lipopolysaccharides
NF-kappa B
Oligodeoxyribonucleotides
Peroxidase
Tumor Necrosis Factor-alpha

Figure

• 

  Figure 1 The time course of acute lung injury (ALI) parameters. The concentrations of tumor necrosis factor (TNF)-α (A) and interleukin (IL)-6 (B) in bronchoalveolar lavage fluid (BALF) and nuclear factor (NF)-κB activity (E) in lung tissue homogenates showed peaks at 6 hours after lipopolysaccharide (LPS) administration. Myeloperoxidase (MPO) activity (C) and ALI score (D) were highest at 36 and 48 hours, respectively (*p<0.05, compared with the control and other time points).

• 

  Figure 2 The effects of nuclear factor (NF)-κB decoy oligodeoxynucleotide (ODN). The concentrations of tumor necrosis factor (TNF)-α (A) and interleukin (IL)-6 (B) and myeloperoxidase (MPO) activity (D) between the lipopolysaccharide (LPS) and LPS+ODN groups were not significantly different. NF-κB activity (C) in the lung tissue homogenates of the LPS+ODN group was significantly lower than the LPS group (*p=0.004, †p>0.05).

• 

  Figure 3 Histopathologic examination (hematoxylin-eosin, ×100) and quantification by acute lung injury (ALI) score. The lipopolysaccharide (LPS) group (B) showed high levels of intra-alveolar exudates, hyaline membrane formation, inflammatory cell infiltration, intra-alveolar hemorrhage, and interstitial edema. These findings were similar with the LPS+oligodeoxynucleotide (ODN) group (C). The ALI score (D) of the LPS+ODN group was not different from the LPS group (*p=0.453).

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