Kidney Week

Abstract: SA-PO767

NAD⁺ Metabolite, 2-Py Has a Potent Anti-Fibrotic and Anti-Inflammatory Activity in the Mouse Unilaterally Ureter-Obstructed Kidney

Session Information

• CKD: Mechanisms - III
  November 09, 2019 | Location: Exhibit Hall, Walter E. Washington Convention Center
  Abstract Time: 10:00 AM - 12:00 PM

Category: Pharmacology (PharmacoKinetics, -Dynamics, -Genomics)

• 1800 Pharmacology (PharmacoKinetics, -Dynamics, -Genomics)

Authors

• Yoshimura, Norito, Shionogi & Co., Ltd., Osaka, Japan

Norito Yoshimura,

• Yamada, Katsutoshi, Shionogi & Co., Ltd., Osaka, Japan

Katsutoshi Yamada,

• Ono, Takashi, Shionogi & Co., Ltd., Osaka, Japan

Takashi Ono,

• Notoya, Mitsuru, Shionogi & Co., Ltd., Osaka, Japan

Mitsuru Notoya,

• Sakamoto, Shingo, Shionogi TechnoAdvance Research Co., Ltd., Osaka, Japan

Shingo Sakamoto,

• Yukioka, Hideo, Shionogi & Co., Ltd., Osaka, Japan

Hideo Yukioka,

• Takahashi, Rina, Keio University, School of Medicine, Tokyo, Japan

Rina Takahashi,

• Kanda, Takeshi, Keio University, School of Medicine, Tokyo, Japan

Takeshi Kanda,

• Wakino, Shu, Keio University, School of Medicine, Tokyo, Japan

Shu Wakino,

• Itoh, Hiroshi, Keio University, School of Medicine, Tokyo, Japan

Hiroshi Itoh,

Background

Renal fibrosis is a common pathogenic feature in chronic kidney diseases. Several studies have suggested NAD⁺ metabolism may be disturbed in the kidney diseases, and we have found that the plasma levels of NAD⁺ metabolites, N-methyl-2-pyridone-5-carboxamide (2-Py) and 4-Py were markedly increased in the unilateral ureter obstruction (UUO) mice and CKD patients. However, the effect of NAD⁺ metabolites on renal fibrosis has not been fully elucidated.

Methods

Effects of 2-Py, 4-Py, and NNO (nicotinamide N-oxide) on TGFβ1 (5 ng/mL)-induced fibrosis-related genes (Col1a1, Col3a1, Col4a1, Acta2) and inflammatory gene (IL-6) were evaluated in rat fibroblast cells (NRK49F) and human tubular cells (HK-2). In vivo study was performed by administering 2-Py (300 mg/kg, bid, plus 5 or 10 mg/mL in the drinking water) to the UUO mice. Body weight and food intake were monitored during this study. At day 7, plasma parameters, gene expression, hydroxyproline content in the kidney were analyzed. Fibrotic area was evaluated using Masson’s trichrome stained sections.

Results

In NRK49F, 2-Py strongly attenuated TGFβ1-induced Col3a1 expression (IC₅₀ 750 µM), and 4-Py showed weak but significant reduction. NNO had no effect. In HK-2, 2-Py prevented TGFβ1-induced Col1a1 expression. 2-Py inhibited TGFβ1-mediated IL-6 gene induction in NRK49F and HK-2. In the UUO mice study, food intake and body weight were not affected by 2-Py treatment, and abnormal behaviors were not observed. At day 7 after UUO treatment, gene expressions of Col1a1, Col3a1, Acta2, IL-6 and TNFα in the kidney were significantly attenuated, and hydroxyproline content and histological fibrotic in the kidney area were significantly lower in the 2-Py-treated group as compared to those in UUO mice with vehicle treatment.

Conclusion

We provide a novel evidence that NAD⁺ metabolite, 2-Py ameliorates renal fibrosis and inflammatory response in renal tubular and fibroblast cells as well as in UUO mice.

Funding

• Commercial Support –