Biomolecules & Therapeutics

The Korean Society of Applied Pharmacology (한국응용약물학회)
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Suppression of Lipopolysaccharide-Induced Inflammatory and Oxidative Response by 5-Aminolevulinic Acid in RAW 264.7 Macrophages and Zebrafish Larvae

  • Ji, Seon Yeong (Anti-Aging Research Center, Dong-eui University) ;
  • Cha, Hee-Jae (Department of Parasitology and Genetics, College of Medicine, Kosin University) ;
  • Molagoda, Ilandarage Menu Neelaka (Department of Marine Life Sciences, Jeju National University) ;
  • Kim, Min Yeong (Anti-Aging Research Center, Dong-eui University) ;
  • Kim, So Young (Anti-Aging Research Center, Dong-eui University) ;
  • Hwangbo, Hyun (Anti-Aging Research Center, Dong-eui University) ;
  • Lee, Hyesook (Anti-Aging Research Center, Dong-eui University) ;
  • Kim, Gi-Young (Department of Marine Life Sciences, Jeju National University) ;
  • Kim, Do-Hyung (Department of Aquatic Life Medicine, Pukyong National University) ;
  • Hyun, Jin Won (Department of Biochemistry, College of Medicine, Jeju National University) ;
  • Kim, Heui-Soo (Department of Biological Sciences, Pusan National University) ;
  • Kim, Suhkmann (Department of Chemistry, Pusan National University) ;
  • Jin, Cheng-Yun (School of Pharmaceutical Sciences, Zhengzhou University) ;
  • Choi, Yung Hyun (Anti-Aging Research Center, Dong-eui University)
  • Received : 2021.02.05
  • Accepted : 2021.03.15
  • Published : 2021.11.01
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Abstract

In this study, we investigated the inhibitory effect of 5-aminolevulinic acid (ALA), a heme precursor, on inflammatory and oxidative stress activated by lipopolysaccharide (LPS) in RAW 264.7 macrophages by estimating nitric oxide (NO), prostaglandin E2 (PGE2), cytokines, and reactive oxygen species (ROS). We also evaluated the molecular mechanisms through analysis of the expression of their regulatory genes, and further evaluated the anti-inflammatory and antioxidant efficacy of ALA against LPS in the zebrafish model. Our results indicated that ALA treatment significantly attenuated the LPS-induced release of pro-inflammatory mediators including NO and PGE2, which was associated with decreased inducible NO synthase and cyclooxygenase-2 expression. ALA also inhibited the LPS-induced expression of pro-inflammatory cytokines, such as tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6, reducing their extracellular secretion. Additionally, ALA abolished ROS generation, improved the mitochondrial mass, and enhanced the expression of heme oxygenase-1 (HO-1) and the activation of nuclear translocation of nuclear factor-E2-related factor 2 (Nrf2) in LPS-stimulated RAW 264.7 macrophages. However, zinc protoporphyrin, a specific inhibitor of HO-1, reversed the ALA-mediated inhibition of pro-inflammatory cytokines production and activation of mitochondrial function in LPS-treated RAW 264.7 macrophages. Furthermore, ALA significantly abolished the expression of LPS-induced pro-inflammatory mediators and cytokines, and showed strong protective effects against NO and ROS production in zebrafish larvae. In conclusion, our findings suggest that ALA exerts LPS-induced anti-inflammatory and antioxidant effects by upregulating the Nrf2/HO-1 signaling pathway, and that ALA can be a potential functional agent to prevent inflammatory and oxidative damage.

Keywords

Acknowledgement

This research was a part of the project titled 'Omics based on fishery disease control technology development and industrialization (20150242)' funded by the Ministry of Oceans and Fisheries, Korea.

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