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Purpose <p>Flos Magnoliae Officinalis (FMO), a traditional Chinese medicinal herb derived from the flower buds of <i>Magnolia officinalis</i>, has long been valued for its anti-inflammatory and antimicrobial properties. However, its non-traditional bioactive components remain underexplored. This study aimed to optimize the recovery of bioactive compounds from FMO using supercritical CO<sub>2</sub> extraction (FMO-SFE) and further evaluate FMO-SFE’s in vitro bioactivity and in vivo efficacy in restoring gut microbiota-immune homeostasis in antibiotic-induced dysbiotic mice.</p> Methods <p>We systematically compared the efficiency of bioactive compound recovery between FMO-SFE and conventional ethanol reflux extraction (FMO-ERP). The chemical profiles of the two extracts were analyzed via UHPLC-Q-TOF-MS/MS metabolomics. Their antioxidant activities were assessed using DPPH, ABTS⁺, and OH⁻ radical scavenging assays. For in vivo experiments, antibiotic-induced dysbiotic mice were administered FMO-SFE at doses of 200–300&#xa0;mg/kg/day. Gut microbiota diversity was evaluated by 16&#xa0;S rRNA sequencing; serum immune markers were profiled via LC-MS-guided cytokine analysis; and biocompatibility was verified through histopathological examination of major organs (heart, liver, spleen, lung, kidney) and detection of key hematological parameters (white blood cells, red blood cells, hemoglobin, platelets).</p> Results <p>Compared with FMO-ERP, FMO-SFE significantly enriched phenolic compounds including magnolol and honokiol, as well as flavonoids like quercetin, with notably enhanced antioxidant activity (<i>p</i> &lt; 0.05). In dysbiotic mice, FMO-SFE effectively restored gut microbiota diversity, normalized the <i>Firmicutes</i>/<i>Bacteroidetes</i> ratio, and increased the abundance of beneficial <i>Lactobacillus</i>. It also repaired ileal mucosal damage and rebalanced the splenic CD3⁺/CD8⁺ T-cell population. Serum cytokine profiling revealed that FMO-SFE dose-dependently reduced IgG, IL-4, and IFN-γ levels while elevating TNF-α, indicating successful restoration of Th1/Th2 immune equilibrium. No adverse histological or hematological changes were observed, confirming the biocompatibility of FMO-SFE at doses up to 300&#xa0;mg/kg.</p> Conclusion <p>These findings position FMO-SFE as a sustainable, LC-MS-validated therapeutic adjuvant for dysbiosis-related disorders. This study therefore links advanced extraction technology directly to functional modulation of the gut microbiota-immune axis, demonstrating a practical model for applying multi-omics approaches to modernize TCM research and development.</p> Graphical Abstract <p></p>

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Supercritical CO2 Extraction Enhances Bioactivity of Flos Magnoliae Officinalis and Restores Gut Microbiota-Immune Homeostasis in Dysbiosis Mice

  • 期刊论文

  • 发表日期：

    2026年3月13日

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  • Yi Jiang,
  • Juan Bai,
  • Yuxin Zhao,
  • Miqi Hu,
  • Zhaohui Li,
  • Dongdong Sun,
  • Weiyun Wang

摘要

Purpose

Flos Magnoliae Officinalis (FMO), a traditional Chinese medicinal herb derived from the flower buds of Magnolia officinalis, has long been valued for its anti-inflammatory and antimicrobial properties. However, its non-traditional bioactive components remain underexplored. This study aimed to optimize the recovery of bioactive compounds from FMO using supercritical CO2 extraction (FMO-SFE) and further evaluate FMO-SFE’s in vitro bioactivity and in vivo efficacy in restoring gut microbiota-immune homeostasis in antibiotic-induced dysbiotic mice.

Methods

We systematically compared the efficiency of bioactive compound recovery between FMO-SFE and conventional ethanol reflux extraction (FMO-ERP). The chemical profiles of the two extracts were analyzed via UHPLC-Q-TOF-MS/MS metabolomics. Their antioxidant activities were assessed using DPPH, ABTS⁺, and OH⁻ radical scavenging assays. For in vivo experiments, antibiotic-induced dysbiotic mice were administered FMO-SFE at doses of 200–300 mg/kg/day. Gut microbiota diversity was evaluated by 16 S rRNA sequencing; serum immune markers were profiled via LC-MS-guided cytokine analysis; and biocompatibility was verified through histopathological examination of major organs (heart, liver, spleen, lung, kidney) and detection of key hematological parameters (white blood cells, red blood cells, hemoglobin, platelets).

Results

Compared with FMO-ERP, FMO-SFE significantly enriched phenolic compounds including magnolol and honokiol, as well as flavonoids like quercetin, with notably enhanced antioxidant activity (p < 0.05). In dysbiotic mice, FMO-SFE effectively restored gut microbiota diversity, normalized the Firmicutes/Bacteroidetes ratio, and increased the abundance of beneficial Lactobacillus. It also repaired ileal mucosal damage and rebalanced the splenic CD3⁺/CD8⁺ T-cell population. Serum cytokine profiling revealed that FMO-SFE dose-dependently reduced IgG, IL-4, and IFN-γ levels while elevating TNF-α, indicating successful restoration of Th1/Th2 immune equilibrium. No adverse histological or hematological changes were observed, confirming the biocompatibility of FMO-SFE at doses up to 300 mg/kg.

Conclusion

These findings position FMO-SFE as a sustainable, LC-MS-validated therapeutic adjuvant for dysbiosis-related disorders. This study therefore links advanced extraction technology directly to functional modulation of the gut microbiota-immune axis, demonstrating a practical model for applying multi-omics approaches to modernize TCM research and development.

Graphical Abstract