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Article

Inhibitory effects of spermidine on lipopolysaccharide‑induced inflammation in RAW 264.7 cells

  • Authors:
    • Hao Yuan
    • Fang Peng
    • Yi-Feng Zhou
    • Si-Xian Wu
    • Zhen-Yi Long
    • Ya-Meng Peng
    • Yi-Zhao Zhou
  • View Affiliations / Copyright

    Affiliations: Clinical Laboratory, Hunan Provincial People's Hospital, The First‑Affiliated Hospital of Hunan Normal University, Changsha, Hunan 410005, P.R. China, Operating Room, Hunan Provincial People's Hospital, The First‑Affiliated Hospital of Hunan Normal University, Changsha, Hunan 410005, P.R. China, Department of Orthopedics, Hunan Provincial People's Hospital, The First‑Affiliated Hospital of Hunan Normal University, Changsha, Hunan 410005, P.R. China
  • Article Number: 124
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    Published online on: March 2, 2026
       https://doi.org/10.3892/etm.2026.13119
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Abstract

Spermidine (SPD) is a naturally occurring polyamine with anti‑inflammatory and antioxidant properties. Given the pivotal role of macrophages in inflammatory pathogenesis (such as rheumatoid arthritis and autoimmune encephalomyelitis), the present study hypothesized that SPD exerts its anti‑inflammatory effects by suppressing macrophage polarization. Briefly, RAW 264.7 cells were activated using lipopolysaccharide (LPS) and were treated with a vehicle control (complete DMEM) or varying SPD concentrations. The ratio of M1 and M2 macrophages in each group was determined using flow cytometry. Inflammatory gene expression and cytokine levels were determined using reverse transcription quantitative PCR and ELISA. The levels of NF‑κB pathway‑associated proteins were measured through western blotting. The findings revealed that the proportion of M1 cells gradually decreased with increasing SPD concentrations. In LPS‑stimulated RAW 264.7 cells, SPD markedly decreased the expression of M1 marker genes and notably increased that of M2 marker genes. Furthermore, SPD decreased IL‑6 levels and increased IL‑10 levels. In addition, the levels of phosphorylated (p)‑p65 and p‑IκBα, which are NF‑κB pathway‑associated proteins, were markedly decreased after 24 h of SPD treatment. Overall, SPD treatment inhibited LPS‑induced M1 polarization, reduced pro‑inflammatory gene/cytokine expression and enhanced anti‑inflammatory markers, potentially through NF‑κB pathway modulation.
View Figures

Figure 1

Impact of different concentrations
and treatment durations of SPD on the viability of RAW 264.7 cells
(n=3; mean ± SD; one-way ANOVA with Tukey's post-hoc test).
***P<0.001, **P<0.01 and
*P<0.05 vs. control group. SPD, spermidine.

Figure 2

Influence of SPD on LPS-induced
polarization of RAW 264.7 cells. Representative flow cytometry
plots of the (A) control, (B) LPS, (C) LPS + 200 µM SPD, (D) LPS +
400 µM SPD and (E) LPS + 800 µM SPD groups. Proportion of (F) M0,
(G) M1 and (H) M2 macrophages (n=3; mean ± SD; one-way ANOVA with
Tukey's post-hoc test). ###P<0.001 and
#P<0.05 vs. control group; ***P<0.001,
**P<0.01 and *P<0.05 vs. LPS group or
as indicated by lines between specific columns. LPS,
lipopolysaccharide; SPD, spermidine.

Figure 3

Effect of SPD on LPS-induced
inflammatory gene expression in RAW 264.7 cells. Expression levels
of (A) Tnfα, (B) Il6, (C) Il1b, (D)
Nos2, (E) Arg1 and (F) Il10 (n=3; mean ± SD;
one-way ANOVA with Tukey's post-hoc test). ###P<0.001
and ##P<0.01 vs. control group;
***P<0.001 and **P<0.01 vs. LPS group.
LPS, lipopolysaccharide; SPD, spermidine; Nos2, nitric oxide
synthase; Arg1, arginase-1.

Figure 4

Influence of SPD on LPS-induced
cytokine secretion in RAW 264.7 cells. Levels of (A) IL-6 and (B)
IL-10 (n=3; mean ± SD; one-way ANOVA with Tukey's post-hoc test).
###P<0.001 and #P<0.05 vs. control
group; ***P<0.001, **P<0.01 and
*P<0.05 vs. LPS group or as indicated by lines
between specific columns. LPS, lipopolysaccharide; SPD,
spermidine.

Figure 5

Effect of SPD on LPS-induced
expression of NF-κB signaling pathway proteins in RAW 264.7 cells.
(A) Representative western blotting images, and relative expression
levels of (B) p-p65/p65 and (C) p-IκBα/IκBα (n=3; mean ± SD;
one-way ANOVA with Tukey's post-hoc test). ###P<0.001
vs. control group; ***P<0.001 vs. LPS group. LPS,
lipopolysaccharide; SPD, spermidine; p-, phosphorylated.
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Copy and paste a formatted citation
Spandidos Publications style
Yuan H, Peng F, Zhou Y, Wu S, Long Z, Peng Y and Zhou Y: Inhibitory effects of spermidine on lipopolysaccharide‑induced inflammation in RAW 264.7 cells. Exp Ther Med 31: 124, 2026.
APA
Yuan, H., Peng, F., Zhou, Y., Wu, S., Long, Z., Peng, Y., & Zhou, Y. (2026). Inhibitory effects of spermidine on lipopolysaccharide‑induced inflammation in RAW 264.7 cells. Experimental and Therapeutic Medicine, 31, 124. https://doi.org/10.3892/etm.2026.13119
MLA
Yuan, H., Peng, F., Zhou, Y., Wu, S., Long, Z., Peng, Y., Zhou, Y."Inhibitory effects of spermidine on lipopolysaccharide‑induced inflammation in RAW 264.7 cells". Experimental and Therapeutic Medicine 31.5 (2026): 124.
Chicago
Yuan, H., Peng, F., Zhou, Y., Wu, S., Long, Z., Peng, Y., Zhou, Y."Inhibitory effects of spermidine on lipopolysaccharide‑induced inflammation in RAW 264.7 cells". Experimental and Therapeutic Medicine 31, no. 5 (2026): 124. https://doi.org/10.3892/etm.2026.13119
Copy and paste a formatted citation
x
Spandidos Publications style
Yuan H, Peng F, Zhou Y, Wu S, Long Z, Peng Y and Zhou Y: Inhibitory effects of spermidine on lipopolysaccharide‑induced inflammation in RAW 264.7 cells. Exp Ther Med 31: 124, 2026.
APA
Yuan, H., Peng, F., Zhou, Y., Wu, S., Long, Z., Peng, Y., & Zhou, Y. (2026). Inhibitory effects of spermidine on lipopolysaccharide‑induced inflammation in RAW 264.7 cells. Experimental and Therapeutic Medicine, 31, 124. https://doi.org/10.3892/etm.2026.13119
MLA
Yuan, H., Peng, F., Zhou, Y., Wu, S., Long, Z., Peng, Y., Zhou, Y."Inhibitory effects of spermidine on lipopolysaccharide‑induced inflammation in RAW 264.7 cells". Experimental and Therapeutic Medicine 31.5 (2026): 124.
Chicago
Yuan, H., Peng, F., Zhou, Y., Wu, S., Long, Z., Peng, Y., Zhou, Y."Inhibitory effects of spermidine on lipopolysaccharide‑induced inflammation in RAW 264.7 cells". Experimental and Therapeutic Medicine 31, no. 5 (2026): 124. https://doi.org/10.3892/etm.2026.13119
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