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. 2025 Mar 25:16:1558916.
doi: 10.3389/fphar.2025.1558916. eCollection 2025.

An active ingredient from the combination of Corydalis Rhizoma and Paeoniae Radix Alba relieves chronic compression injury-induced pain in rats by ameliorating AR/Mboat2-mediated ferroptosis in spinal cord neurons

Ze-Ming Wang #  1 , Xiao-Hong Wei #  2 , Gui-Yang Xia  2 , Lin-Nan Zhou  1 , Jin-Yu Li  2 , Sheng Lin  2
Affiliations

An active ingredient from the combination of Corydalis Rhizoma and Paeoniae Radix Alba relieves chronic compression injury-induced pain in rats by ameliorating AR/Mboat2-mediated ferroptosis in spinal cord neurons

Ze-Ming Wang et al. Front Pharmacol. .

Abstract

Introduction: A combination of Corydalis Rhizoma (the dried tuber of Corydalis yanhusuo W.T. Wang) and Paeoniae Radix Alba (the root of Paeonia lactiflora Pall.) has been traditionally employed for analgesia. However, the underlying pharmacological mechanisms have not been clarified. The aim of the present study was to investigate the anti-inflammatory and analgesic effects of YB60, the 60% ethanol elution fraction derived from the combination of Corydalis Rhizoma and Paeoniae Radix Alba, and the explore the underlying mechanism.

Methods: Lipopolysaccharide-induced cellular inflammation model and chronic compression injury (CCI) rat model were used to study the anti-inflammatory and analgesic effects of YB60. Proteomics and molecular biology experiments were applied to explore the potential analgesic mechanism of YB60.

Results: The results demonstrated that YB60 significantly decreased inflammatory cytokine levels both in cellular models and rat serum, while concurrently elevating pain thresholds in CCI rats. Proteomic analysis indicated that YB60 could upregulate the expression of Membrane Bound O-Acyltransferase Domain Containing 2 (Mboat2), a newly confirmed marker of ferroptosis. Furthermore, YB60 prevented ferroptosis in the spinal cords of CCI rats. Western blotting and immunofluorescent dual staining further revealed that YB60 increased the expression of Mboat2 and its upstream signaling molecule Androgen receptor (AR). Results in PC12 cells in vitro showed that YB60 reversed the downregulation of AR and Mboat2, and ameliorated ferroptosis induced by Erastin, while knockdown of AR eliminated the above effects of YB60.

Conclusion: These findings indicated that YB60 exerted its analgesic effect by inhibiting ferroptosis in spinal cord neurons via modulation of the AR/Mboat2 pathway.

Keywords: AR; Mboat2; analgesic effect; anti‐inflammatory; chronic compression injury; chronic pain; extract of Corydalis Rhizoma and Paeoniae Radix Alba; ferroptosis.

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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
YB60 alleviated inflammation induced by LPS in RAW 264.7 and BV2 cells. A-C. The effects of YB60 on TNF-α (A), IL-6 (B), and IL-1β (C) contents in RAW 264.7 cells. D-F. Effects of YB60 on TNF-α (D), IL-6 (E), and IL-1β (F) levels in BV2 cells. The error bars represent the SD. ** P < 0.01 vs. the control group; ## P < 0.01 vs. the LPS group.
FIGURE 2
FIGURE 2
YB60 has strong anti-inflammatory and pain-relieving effects on CCI rats. A-C. Serum levels of TNF-α (A), IL-6 (B), and IL-1β (C) in each group of rats. (D) MWT in each group of rats. (E) TWL in each group of rats. The error bars represent the SD. ** P < 0.01 vs. the sham group. # P < 0.05 vs. the CCI group. ## P < 0.01 vs. the CCI group.
FIGURE 3
FIGURE 3
YB60 modulates ferroptosis in spinal cord of CCI rats. (A) TEM images of the L4‒L6 spinal cord dorsal horns of Sham, CCI, and CCI + YB60-H rats, with arrows pointing to mitochondria displaying ferroptosis features. B-D. Contents of iron ions (B), MDA (C) and ROS (D) in various groups, n = 6. The error bars represent the SD. ** P < 0.01 compared with the Sham group. # P < 0.05 compared with the CCI group. ## P < 0.01 compared with the CCI group.
FIGURE 4
FIGURE 4
TMT proteomics analysis of the L4‒L6 spinal cord segments in CCI rats. (A) Differential protein expression. The volcano plot displays log2 (FC) on the horizontal axis, with values further from zero indicating greater differential expression, right indicating upregulation, and left indicating downregulation. The vertical axis shows -log10(P value), with higher values indicating greater significance. Red and blue dots indicate upregulated and downregulated proteins, respectively, with deeper colors indicating more significant changes, whereas gray dots indicate proteins with p values ≥0.05. (B) Venn diagram of the differentially expressed proteins. Different colors denote different groups, with the numbers indicating the count of intersecting and unique proteins for each group. (C) Heatmap of differential protein clustering. Proteins are clustered by expression level, with red indicating higher expression and blue indicating lower expression. Each row represents the expression level of each protein across different groups. (D) KEGG pathway analysis of intersecting proteins. (E) The protein level of Mboat2 in the Sham, CCI and CCI + YB60-H groups. ** P < 0.01.
FIGURE 5
FIGURE 5
YB60 modulates ferroptosis and the AR/Mboat2 axis in Spinal Cord Neurons. (A) Fluorescence double-label staining of AR (green) and c-Fos (red) in the Sham (a1, a4), CCI (a2, a5) and CCI + YB60-H (a3, a6) groups. DAPI (blue) staining highlights nuclei. Scale bars: 500 μm (a1- a3); 100 μm (a4- a6). (B) Fluorescence double-label staining of Mboat2 (green) and c-Fos (red) in the sham (b1, b4), CCI (b2, b5) and CCI + YB60-H (b3, b6) groups. DAPI (blue) staining highlights nuclei. Scale bars: 500 μm (b1- b3); 100 μm (b4- b6). (C) Expression levels of AR and Mboat2. (D-E) Semiquantitative analysis of AR (D) and Mboat2 (E), n = 3. The error bars represent the SD. ** P < 0.01.
FIGURE 6
FIGURE 6
YB60 inhibited Erastin-induced ferroptosis by regulating AR/Mboat2 in PC12 cells. (A) Expression levels of AR and Mboat2 in PC12 cells. (B-C) Semiquantitative analysis of AR (B) and Mboat2 (C), n = 3. D-F. Contents of iron ions (D), ROS (E) and MDA (F) in various groups, n = 6. (G) Viability of PC12 cells. The error bars represent the SD. ** P < 0.01.
FIGURE 7
FIGURE 7
YB60 inhibited ferroptosis in spinal cord neurons by increasing the expression of AR and Mboat2.
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References

    1. Arafat M. A. M., Khalil M. N. A., Mohamed O. G., Abd EL-Ghafar O. A. M., Tripathi A., Mahrous E. A., et al. (2023). Vetiver aerial parts and roots ameliorate rheumatoid arthritis in complete Freund's adjuvant rat model, a phytochemical profiling and mechanistic study. J. Ethnopharmacol. 317, 116764. 10.1016/j.jep.2023.116764 - DOI - PubMed
    1. Bennett G. J., Xie Y. K. (1988). A peripheral mononeuropathy in rat that produces disorders of pain sensation like those seen in man. Pain 33, 87–107. 10.1016/0304-3959(88)90209-6 - DOI - PubMed
    1. Bersuker K., Hendricks J. M., Li Z., Magtanong L., Ford B., Tang P. H., et al. (2019). The CoQ oxidoreductase FSP1 acts parallel to GPX4 to inhibit ferroptosis. Nature 575, 688–692. 10.1038/s41586-019-1705-2 - DOI - PMC - PubMed
    1. Biglione B., Gitin A., Gorelick P. B., Hennekens C. (2020). Aspirin in the treatment and prevention of migraine headaches: possible additional clinical options for primary healthcare providers. Am. J. Med. 133, 412–416. 10.1016/j.amjmed.2019.10.023 - DOI - PubMed
    1. Breault É., Desgagné M., Neve J., CôTé J., Barlow T. M. A., Ballet S., et al. (2024). Multitarget ligands that comprise opioid/nonopioid pharmacophores for pain management: current state of the science. Pharmacol. Res. 209, 107408. 10.1016/j.phrs.2024.107408 - DOI - PubMed

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