Journal of Veterinary Science

The Korean Society of Veterinary Science (대한수의학회)
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Anti-inflammatory effect of sulforaphane on LPS-stimulated RAW 264.7 cells and ob/ob mice

  • Ranaweera, Sachithra S. (College of Veterinary Medicine, Jeju National University) ;
  • Dissanayake, Chanuri Y. (College of Veterinary Medicine, Jeju National University) ;
  • Natraj, Premkumar (College of Veterinary Medicine, Jeju National University) ;
  • Lee, Young Jae (College of Veterinary Medicine, Jeju National University) ;
  • Han, Chang-Hoon (College of Veterinary Medicine, Jeju National University)
  • Received : 2020.09.18
  • Accepted : 2020.10.28
  • Published : 2020.11.30
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Abstract

Background: Sulforaphane (SFN) is an isothiocyanate compound present in cruciferous vegetables. Although the anti-inflammatory effects of SFN have been reported, the precise mechanism related to the inflammatory genes is poorly understood. Objectives: This study examined the relationship between the anti-inflammatory effects of SFN and the differential gene expression pattern in SFN treated ob/ob mice. Methods: Nitric oxide (NO) level was measured using a Griess assay. The inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression levels were analyzed by Western blot analysis. Pro-inflammatory cytokines (tumor necrosis factor [TNF]-α, interleukin [IL]-1β, and IL-6) were measured by enzyme-linked immunosorbent assay (ELISA). RNA sequencing analysis was performed to evaluate the differential gene expression in the liver of ob/ob mice. Results: The SFN treatment significantly attenuated the iNOS and COX-2 expression levels and inhibited NO, TNF-α, IL-1β, and IL-6 production in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. RNA sequencing analysis showed that the expression levels of 28 genes related to inflammation were up-regulated (> 2-fold), and six genes were down-regulated (< 0.6-fold) in the control ob/ob mice compared to normal mice. In contrast, the gene expression levels were restored to the normal level by SFN. The protein-protein interaction (PPI) network showed that chemokine ligand (Cxcl14, Ccl1, Ccl3, Ccl4, Ccl17) and chemokine receptor (Ccr3, Cxcr1, Ccr10) were located in close proximity and formed a "functional cluster" in the middle of the network. Conclusions: The overall results suggest that SFN has a potent anti-inflammatory effect by normalizing the expression levels of the genes related to inflammation that were perturbed in ob/ob mice.

Keywords

Acknowledgement

This study was supported by the program of supporting Promising Small and Medium Industry (Grant No: C0518751) funded by the Korea Small and Medium Business Administration in 2017. The study was also supported by the 2019 scientific promotion program funded by Jeju National University.

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