Korean J Urol.  2007 Aug;48(8):815-825. 10.4111/kju.2007.48.8.815.

The Effect of Ring-type NF-kappa B(NF-kB) Decoy Oligodeoxynucleotide on the Kidney for an Experimental Unilateral Ureteral Obstruction in Mice

• Kwon YG
• Lee YG
• Kang I
• Lee ES
• Park KK
• Keum YS
• Kim KH
• Park JS

• KMID: 2061275
• DOI: http://doi.org/10.4111/kju.2007.48.8.815

Abstract

PURPOSE
The transcription factor, NF-kB, is important in the coordinated expressions of various pro-inflammatory and adhesion molecules. Our hypothesis is that inhibiting the action of NF-kB using a synthetic decoy oligodeoxynucleotide(ODN) can block the underlying inflammatory response in glomerulonephritis.
MATERIALS AND METHODS
Forty two male C57BL6 mice, weighting 25g, were divided into four groups; in group 1, 2 mice were used as normal controls; in group 2, a unilateral ureteral obstruction(UUO) was induced in 12 mice; in group 3, 20 UUO mice were treated using ring-type NK-kB decoy ODN; and in group 4, 8 UUO mice were treated using scramble type NF-kB decoy ODN. The mice were killed 1, 3, 5 and 7 days after the NF-kB decoy ODN injections, and the blood urea nitrogen(BUN), and expressions of tumor necrosis factor-alpha (TNF-alpha), interleukin-beta(IL-beta), fibronectin, vascular cell adhesion molecule(VCAM) and monocyte chemotactic protein-1(MCP-1), as well as the histopathological findings, analyzed in each group.
RESULTS
The serum levels of BUN, TNF-alpha, IL-beta, fibronectin, VCAM and MCP-1 were increased in group 2, but decreased in group 3. The histopathological findings of the kidneys in group 3 were most similar to those found in the control(group 1).
CONCLUSIONS
NF-kB decoy ODN treatment substantially inhibited the disease, with reductions in the histological damage and the renal expressions of inflammatory cytokines.

Keyword

NF-kappa B; Transcription factors; Ureteral obstruction

MeSH Terms

Animals
Cell Adhesion
Cytokines
Fibronectins
Glomerulonephritis
Humans
Kidney*
Male
Mice*
Monocytes
NF-kappa B
Transcription Factors
Tumor Necrosis Factor-alpha
Urea
Ureter*
Ureteral Obstruction*
Cytokines
Fibronectins
NF-kappa B
Transcription Factors
Tumor Necrosis Factor-alpha
Urea

Figure

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  Fig. 1. Fluorescent microscopic view of the transfected NF-kappa B (NF-kB) decoy oligodeoxynucleotide (ODN) into the renal tubular cells.

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  Fig. 2. Structure and molecular stability of a Sp1 decoy oligode-oxynucleotide (ODN). (A) Structure of a ring-type NF-kappa B (NF-kB) decoy ODN, the NF-kB binding site is underlined, (B) stability of a NF-kB decoy ODN in the presence of exonuclease III (Exo III). Linear NF-kB ODNs were completely digested after incubation for 24 hr in the presence of Exo III.

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  Fig. 3. Analysis of the effect of ring-type NF-kappa B (NF-kB) decoy oligodeoxynucleotide (ODN) on the function of the kidney using ELISA; showing biochemical analyses of the blood urea nitrogen (A), the serum levels of tumor necrosis factor-α (B) and interleukin-1β(C) from UUO and UUO mice treated with decoy ODN. UUO (n=12): unilateral ureteral obstruction, UUO+NF-kB (n=20): UUO mice treated with a decoy ODN injection. The UUO+NF-kB group was found to be significantly statistically different to the UUO group using the chi-square test (p＜0.05).

• 

  Fig. 4. Ring-type NF-kappa B (NF-kB) decoy oligodeoxynucleo-tide (ODN) effectively inhibits the expressions of fibronectin, vascular cell adhesion molecule (VCAM) and monocyte chemotactic protein-1 (MCP-1) in the kidneys treated with NF-kB decoy ODN. Protein expressions of fibronectin (A), VCAM(B) and MCP-1 (C) were determined by Western blotting. The signal intensity was quantified using a densitometric analysis. D: day, NC: negative control, Control (n=2): unilateral ureteral obstruction (UUO), SM (n=8): scrambled decoy ODN, WT (n=20): wild type decoy ODN. The WT NF-kB group was found to be significantly statistically different to the UUO group using the chi-square test (p＜0.05).

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  Fig. 5. The microscopic findings of the kidneys after UUO shows the infiltration of inflammatory cells and mild fibrosis, with tubular atrophy, compared with the negative control (A). The renal morphology of rats was determined using HE-staining at 3 (B), 5 (C) and 7 days (D) after the operation. UUO: unilateral ureteral obstruction.

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  Fig. 6. Treatment with ring-type NF-kappa B (NF-kB) decoy oligodeoxynucleotide (ODN) suppresses interstitial inflammation and fibrosis. The renal morphology was determined using HE-staining at 1 (A), 3 (B) and 7 (C) days after the transfection of NF-kB decoy ODN. Left: kidney of UUO (n=12), Right: kidney of UUO treated with NF-kB decoy ODN (n=20), UUO: unilateral ureteral obstruction.

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  Fig. 7. Ring-type NF-kappa B (NF-kB) decoy oligodeoxynucleo-tide (ODN) prevents the UUO-induced expressions of NF-kB target genes 7 days after a ureteral obstruction. The kidney sections of NF-kB decoy ODN-treated rats were stained by antibodies for TNF-α. (A) Negative control (n=2), (B) kidney of UUO (n=12), (C) kidney of UUO treated with scrambled NF-kB decoy ODN (n=8), (D) kidney of UUO treated with NF-kB decoy ODN (n=20). UUO: unilateral ureteral obstruction.

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  Fig. 8. Ring-type NF-kappa B (NF-kB) decoy oligodeoxynucleo-tide (ODN) prevents the UUO-induced expressions of NF-kB target genes 7 days after a ureteral obstruction. The kidney sections of NF-kB decoy ODN-treated rats were stained by antibodies for IL-1β. (A) Negative control (n=2), (B) kidney of UUO (n=12), (C) kidney of UUO treated with scrambled NF-kB decoy ODN (n=8), (D) kidney of UUO treated with NF-kB decoy ODN (n=20). UUO: unilateral ureteral obstruction, IL: interleukin.

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  Fig. 9. Ring-type NF-kappa B (NF-kB) decoy oligodeoxynucleoti-de (ODN) prevents the UUO-induced expressions of NF-kB target genes 7 days after a ureteral obstruction. The kidney sections of NF-kB decoy ODN-treated rats were stained by antibodies for fibronectin. (A) Negative control (n=2), (B) kidney of UUO (n=12), (C) kidney of UUO treated with scrambled NF-kB decoy ODN (n=8), (D) kidney of UUO treated with NF-kB decoy ODN (n=20). UUO: unilateral ureteral obstruction.

• 

  Fig. 10. Ring-type NF-kappa B (NF-kB) decoy oligodeoxynucleo-tide (ODN) prevents the UUO-induced expressions of NF-kB target genes 7 days after a ureteral obstruction. The kidney sections of NF-kB decoy ODN-treated rats were stained by antibodies for VCAM. (A) Negative control (n=2), (B) kidney of UUO (n=12), (C) kidney of UUO treated with scrambled NF-kB decoy ODN (n=8), (D) kidney of UUO treated with NF-kB decoy ODN (n=20). UUO: unilateral ureteral obstruction.

• 

  Fig. 11. Ring-type NF-kappa B (NF-kB) decoy oligodeoxynucleo-tide (ODN) prevents the UUO-induced expressions of NF-kB target genes 7 days after ureteral obstruction. The kidney sections of NF-kB decoy ODN-treated rats were stained by antibodies for monocyte chemotactic protein-1 (MCP-1). (A) Negative control (n=2), (B) kidney of UUO (n=12), (C) kidney of UUO treated with scrambled NF-kB decoy ODN (n=8), (D) kidney of UUO treated with NF-kB decoy ODN (n=20). UUO: unilateral ureteral obstruction.

Reference

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