Nutr Res Pract.  2025 Feb;19(1):143-153. 10.4162/nrp.2025.19.1.143.

Anti-inflammatory effects of rutin in lipopolysaccharide-stimulated canine macrophage cells

• Jang H
• Han SC
• Lee J
• Shin HY
• Hwang JH
• Ha JH

• KMID: 2565391
• DOI: http://doi.org/10.4162/nrp.2025.19.1.143

Abstract

BACKGROUND/OBJECTIVES
Inflammatory responses are key pathological factors in various canine diseases, making the control of inflammatory responses vital for canine health. This study examined the anti-inflammatory effects of rutin on DH82 cells, a type of canine macrophage, against lipopolysaccharide (LPS)-induced inflammatory responses.
MATERIALS/METHODS
The inflammatory in vitro experimental model was established by stimulating canine macrophage DH82 cells with LPS. To evaluate the inflammationpreventative effects of rutin, analyses were conducted using enzyme-linked immunosorbent assay, western blot, and real-time quantitative reverse transcription polymerase chain reaction.
RESULTS
Rutin inhibited the LPS-induced increase in the protein and gene levels of proinflammatory cytokines (interleukin [IL]-1β, IL-6, tumor necrosis factor-α), while antiinflammatory cytokines (IL-10, transforming growth factor-β1) levels remained unchanged. Furthermore, rutin suppressed the LPS-induced activation of phosphorylated extracellular signal-regulated kinase, Jun N-terminal kinase, inhibitor of nuclear factor kappa B, and nuclear factor kappa B (NF-κB) in DH82 cells.
CONCLUSION
Rutin exerts anti-inflammatory effects by inhibiting the mitogen-activated protein kinase-NF-κB signaling pathway and reducing the production of pro-inflammatory cytokines in DH82 cells.

Keyword

Dogs; macrophages; inflammation; rutin

Figure

• 

  Fig. 1 The effect of rutin on LPS-induced pro/anti-inflammatory cytokines in DH82 cells. After 24 h of rutin treatment (0–40 μM), DH82 cells are exposed to LPS (0.1 μg/mL) for 6 h. (A) TNF-α levels, (B) IL-10 levels, (C) TNF-α/IL-10 ratio. Values are expressed as the mean ± SD.LPS, lipopolysaccharide; TNF-α, tumor necrosis factor-α; IL-10, interleukin 10.a,b,c,dMeans with different letters indicate significant differences (P < 0.05) as determined one-way analysis of variance, followed by Tukey’s post hoc test.

• 

  Fig. 2 The effect of rutin on LPS-induced proinflammatory gene expression, MAPK, and NF-κB activation in DH82 cells. After 24 h of rutin treatment (0–40 μM), DH82 cells are exposed to LPS (0.1 μg/mL) for 5 or 10 min. (A) Representative western blot images, (B) phosphorylated IκB levels, (C) phosphorylated NF-κB levels, (D) phosphorylated ERK levels, (E) phosphorylated JNK levels. Protein levels are adjusted relative to the GAPDH reference standard. Values are expressed as the mean ± SD.LPS, lipopolysaccharide; JNK, Jun N-terminal kinase; MAPK, mitogen-activated protein kinase; NF-κB, nuclear factor kappa B; IκB, inhibitor of nuclear factor kappa B; ERK, extracellular signal-regulated kinase; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; p, phosphorylation; t, total.a,b,cMeans with different letters indicate significant differences (P < 0.05) as determined one-way analysis of variance, followed by Tukey’s post hoc test.

• 

  Fig. 3 The effect of rutin on LPS-induced mRNA inflammation-related expression in DH82 cells. After 24 h of rutin treatment (0–40 μM), DH82 cells are exposed to LPS (0.1 μg/mL) for 3 h. (A) IL-6 levels, (B) IL-1β levels, (C) TNF-α levels, (D) TGF-β1 levels, (E) IL-10 levels. The mRNA levels are adjusted relative to the GAPDH reference standard. Values are expressed as the mean ± SD.LPS, lipopolysaccharide; IL, interleukin; TNF, tumor necrosis factor; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; NS, not statistically significant.a,b,cMeans with different letters indicate significant differences (P < 0.05) as determined one-way ANOVA, followed by Tukey’s post hoc test.

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