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. 2025 Feb 4:12:1539448.
doi: 10.3389/fvets.2025.1539448. eCollection 2025.

Normal butanol fraction of Polygonum hydropiper L. flavonoids reduces inflammation caused by PCV2 infections in cell and mouse models

Yu-Heng Wei  1 , Shu-Mian Zhou  1   2   3 , Wen Zhao  1 , Qi Chen  1 , Qiu-Hua Wang  1   3 , Mei-Ling Yu  1 , Ying-Yi Wei  1 , Ting-Jun Hu  1   2   3
Affiliations

Normal butanol fraction of Polygonum hydropiper L. flavonoids reduces inflammation caused by PCV2 infections in cell and mouse models

Yu-Heng Wei et al. Front Vet Sci. .

Abstract

Introduction: The normal butanol fraction of Polygonum hydropiper L. flavonoids (FNB) exhibits significant anti-inflammatory effects. This study investigated FNB's impact on inflammatory responses induced by Porcine circovirus type 2 (PCV2) in cell and mouse models.

Methods: An inflammatory model was established in RAW264.7 cells infected with varying PCV2 concentrations. And assigning both RAW264.7 cells and 108 SPF-grade KM mice to Control, PCV2, Rutin, and various dosages of FNB groups. Inflammatory factors such as Monocyte Chemoattractant Protein-1 (MCP-1), interleukin-6 (IL-6), IL-8, IL-10, Tumor Necrosis Factor-alpha (TNF-α), Reactive Oxygen Species (ROS), and Nitric Oxide (NO) were quantified using ELISA, RT-qPCR and immunohistochemistry.

Results: Results showed that a PCV2 titer of 104.5 TCID50/0.1 mL when applied to RAW264.7 cells effectively established an in vitro inflammatory model at 12 and 24 h post-infection. Following PCV2 infection, all the inflammatory factors displayed a significant increased both in culture supernatant and intracellular mRNA expression levels (p < 0.05 or p < 0.01), but these levels were reduced by FNB treatment (p < 0.05 or p < 0.01). In mouse sera post-PCV2 infection also showed elevated levels of IL-6, IL-8 IL-10, TNF-α, and MCP-1 (p < 0.05 or p < 0.01). Additionally, mRNA and protein levels for TNF-α, IL-8, IL-10, IL-6, and iNOS rose significantly in lung tissues (p < 0.01) but decreased with FNB treatment (p < 0.05 or p < 0.01).

Discussion: These findings suggest that FNB reduces inflammatory factor production and modulates the inflammatory response triggered by PCV2 infection, potentially enhancing host resistance against it.

Keywords: RAW264.7 cells; inflammatory response; model establishment; normal butanol fraction of Polygonum hydropiper L. flavonoids; porcine circovirus type 2.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
(A–G) Effects of PCV2 at different PCV2 dilutions on MCP-1, TNF-α, IL-6, IL-10, IL-8, ROS and NO level (mean ± SD, n = 4). *p < 0.05, **p < 0.01 vs. Control group.
Figure 2
Figure 2
Effects of varying concentrations of FNB on RAW264.7 cell viability (mean ± SD, n = 4). **p < 0.01 vs. Control group.
Figure 3
Figure 3
(A–G) The impact of FNB as an intervention on inflammatory cytokine levels (mean ± SD, n = 4). *p < 0.05, **p < 0.01 vs. Control group. #p < 0.05, ##p < 0.01 vs. PCV2 group.
Figure 4
Figure 4
Impact of FNB on inflammatory cytokine mRNA expression levels (mean ± SD, n = 4). **p < 0.01 vs. Control group. #p < 0.05, ##p < 0.01 vs. PCV2 group.
Figure 5
Figure 5
(A–E) Effects of FNB on inflammatory cytokine levels in mouse sera after PCV2 infection (mean ± SD, n = 6). *p < 0.05, **p < 0.01 vs. Control group. #p < 0.05, ##p < 0.01 vs. PCV2 group.
Figure 6
Figure 6
(A–F) Impact of FNB on inflammatory cytokine mRNA expression in mouse lungs following PCV2 infection (mean ± SD, n = 6). **p < 0.01 vs. Control group. ##p < 0.01 vs. PCV2 group.
Figure 7
Figure 7
(A, B) Effects of FNB on inflammatory cytokine protein levels in mouse lungs following PCV2 infection (mean ± SD, n = 6). **p < 0.01 vs. Control group. ##p < 0.01 vs. PCV2 group.
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