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

Young Gun Kwon; Young Guk Lee; Il Kang; Eun Suk Lee; Kwan Kyu Park; Yoon Seop Keum; Kyung Hyun Kim; Jae Shin Park

doi:10.4111/kju.2007.48.8.815

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Kwon, Lee, Kang, Lee, Park, Keum, Kim, and Park: The Effect of Ring-type NF-kappa B(NF-kB) Decoy Oligodeoxynucleotide on the Kidney for an Experimental Unilateral Ureteral Obstruction in Mice

Original Article

Korean J Urol 2007;48(8):815-825.

Published online: 20 January 2007

DOI: https://doi.org/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

Young Gun Kwon, Young Guk Lee, Il Kang, Eun Suk Lee, Kwan Kyu Park¹, Yoon Seop Keum¹, Kyung Hyun Kim¹, Jae Shin Park

From the Department of Urology, Daegu Catholic University College of Medicine, Daegu, Korea

¹From the Department of Pathology, Daegu Catholic University College of Medicine, Daegu, Korea

대구가톨릭대학교 의과대학 비뇨기과학교실 대구광역시 남구 대명 4동 3056-6 ? 705-034 TEL: 053-650-4662 FAX: 053-623-4660 E-mail: jspark@cu.ac.kr

Received 30 October 2006 Accepted 5 June 2007

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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-α(TNF-α), interleukin-β (IL-β), 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-α, IL-β, 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.

Keywords: NF-kappa B, Transcription factors, Ureteral obstruction

References

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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.

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.

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).

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.

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.

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.

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.

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.

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