{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T16:18:35Z","timestamp":1792685915764,"version":"4.5.0"},"reference-count":54,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,8,21]],"date-time":"2025-08-21T00:00:00Z","timestamp":1755734400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec><jats:title>Purpose<\/jats:title><jats:p>This study aimed to investigate whether Jianpi-Zishen Formula (JPZS) modulates the Treg\/Th17 balance in MRL\/lpr mice through regulation of DNA methyltransferase 1 (DNMT1)-mediated forkhead box P3 (Foxp3) methylation, and to elucidate its potential mechanism for improving immune homeostasis in systemic lupus erythematosus (SLE).<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Forty-eight female MRL\/lpr mice were randomized into six groups (n=8\/group): JPZS (low\/medium\/high doses), 5-aza-CdR (DNMT inhibitor), DC_517 (DNMT1 inhibitor), and model control. Eight C57BL\/6 mice served as healthy controls. The mice were subjected to the corresponding intervention measures for eight weeks. The impact of JPZS on the disease progression of MRL\/lpr mice was evaluated using enzyme-linked immunosorbent assay (ELISA) and serum biochemical parameters. Moreover, immunofluorescence staining and flow cytometry were employed to investigate alterations in the proportions of Tregs and Th17 cells. CD4<jats:sup>+<\/jats:sup> T cells were isolated from the spleen for subsequent investigation, including quantitative real-time PCR, western blotting, and determination of DNA methylation levels. Furthermore, the enzymatic activity of CD4<jats:sup>+<\/jats:sup> T cell-specific DNA methyltransferases was quantified using an EpiQuik DNMT detection kit.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>JPZS significantly improved the disease development of MRL\/lpr mice in a dose-dependent manner. Flow cytometry and immunofluorescence indicated JPZS promoted Treg\/Th17 rebalancing. Research has found that <jats:italic>Foxp3<\/jats:italic> is at a high methylation level in CD4<jats:sup>+<\/jats:sup> T cells of the model group, and the transcription level of <jats:italic>Foxp3<\/jats:italic> mRNA is downregulated; JPZS can downregulate <jats:italic>Foxp3<\/jats:italic> methylation levels of CD4<jats:sup>+<\/jats:sup> T cells in the model group. Further research has found that the level of <jats:italic>Foxp3<\/jats:italic> methylation is closely related to Dnmt1 enzyme activity, and JPZS can downregulate Dnmt1 enzyme activity, thereby upregulating the transcription level of <jats:italic>Foxp3<\/jats:italic> mRNA.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>JPZS may restore Treg\/Th17 balance in SLE via DNMT1-regulated <jats:italic>Foxp3<\/jats:italic> demethylation, suggesting an epigenetic mechanism for its immunomodulatory effects.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fimmu.2025.1631631","type":"journal-article","created":{"date-parts":[[2025,8,21]],"date-time":"2025-08-21T04:12:18Z","timestamp":1755749538000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":3,"title":["Promotion of Treg\/Th17 balance in MRL\/lpr mice by Jianpi-Zishen Formula via modulation of DNMT1-mediated Foxp3 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