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. 2026 Feb 2;21:59. doi: 10.1186/s13020-026-01338-8

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© The Author(s) 2026

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 6 — Identification of a miRNA–FOXO3–CXCR5 regulatory axis. A Volcano plot of microRNA expression profiles in the Sp5C region, showing differentially expressed microRNAs between experimental groups. B Quantitative PCR analysis of miR-12179-5p (R² = 0.9155, Con vs. IS, p = 0.0003; IS vs. IS + EA, p = 0.0077), miR-23b-5p (R² = 0.9204, Con vs. IS, p = 0.0004; IS vs. IS + EA, p = 0.0153), miR-214-5p (R² = 0.8581, Con vs. IS, p = 0.0017; IS vs. IS + EA, p = 0.0324), and miR-6239 (R²= 0.9721, Con vs. IS, p < 0.0001; IS vs. IS + EA, p = 0.0002) expression in the Sp5C across the indicated groups. C Quantitative PCR analysis of CXCR5 mRNA expression in Neuro-2a cells following transfection with the indicated microRNA mimics or negative control (miR-12179-5p vs. NC, R² = 0.9215, P = 0.0024; miR-23b-5p vs. NC, R² = 0.8662, P = 0.007; miR-214-5p vs. NC, R² = 0.9484, P = 0.001; miR-6239 vs. NC, R²= 0.9071, P = 0.0033). D Venn diagram showing the overlap of predicted target genes for miR-23b-5p, miR-214-5p, and miR-6239 based on TargetScan and miRanda analyses. E Dual-luciferase reporter assay assessing the effect of the indicated microRNA mimics on CXCR5 3′ untranslated region (3′UTR)–driven luciferase activity (R² = 0.5997, miR-214-5p vs. NC, p = 0.212; miR-23b-5p vs. NC, p = 0.3633; miR-6239 vs. NC, p = 0.8513). F Dual-luciferase reporter assay evaluating the effect of the indicated microRNA mimics on CXCR5 promoter–driven luciferase activity (R² = 0.9929, miR-23b-5p vs. NC, p < 0.0001; miR-214-5p vs. NC, p < 0.0001; miR-6239 vs. NC, p < 0.0001). G KEGG pathway enrichment analysis of the shared predicted target genes identified in D. H Dual-luciferase reporter assay examining the effect of the indicated microRNA mimics on FOXO3 3′UTR–driven luciferase activity (R² = 0.9363, miR-23b-5p vs. NC, p < 0.0006; miR-214-5p vs. NC, p < 0.0001; miR-6239 vs. NC, p < 0.0002). I Quantitative PCR analysis of FOXO3 mRNA expression in Neuro-2a cells following transfection with the indicated microRNA mimics or negative control (miR-23b-5p vs. NC, R²= 0.9893, P < 0.0001; miR-214-5p vs. NC, R² = 0.9885, P < 0.0001; miR-6239 vs. NC, R²= 0.9683, P = 0.0004. J Western blot analysis of FOXO3 protein expression in the Sp5C across the indicated experimental groups (R² = 0.9945, Con vs. IS, P < 0.0001; IS vs. IS + EA, P < 0.0001). Data are presented as mean ± SD.