Latex-bearing plants have gained global attention, but few studies have been reported on Codonopsis pilosula (Franch.) Nannf.( C. pilosula ) latex. In this research, we established a UHPLC-MS/MS method to investigate the constituents in C. pilosula latex. Analysis revealed that the latex of C. pilosula contains alkaloids, terpenoids, and fatty acids, with fatty acids being one of the primary metabolites distinguishing the latex from above-ground (DS) and below-ground (DX) parts of C. pilosula. To evaluate the quality of C. pilosula latex, an approach of dispersive liquid-liquid microextraction with solidification of the floating organic drop (DLLME-SFO) methodology was developed to quantitatively analyze the contents of three fatty acids (palmitic, linoleic, and stearic) in the latex of C. pilosula . The ideal extraction conditions involve injecting 20 μL of dodecanol extractant into a 5 mL 10% NaCl solution at pH 3.0, followed by 20 min at 700 rpm. The three components mentioned above have enrichment coefficients ranging from 48 to 50. The quantification limits were between 0.23 and 0.25 μg/mL, and three components demonstrated good linearity in their respective linear ranges (R 2 > 0.995). The satisfied precision and recovery rates were 1.26 to 8.05% and 92.38 to 101.53%, respectively. By quantitatively analyzing the three fatty acid contents in the latex of C. pilosula , the results showed that palmitic and linoleic acids were able to successfully distinguish the latex of C. pilosula from different parts, which laid the foundation for the quality evaluation of C. pilosula , and provided a reference for the utilization of C. pilosula , a medicinal food source.
Background Dextran sulfate sodium-induced ulcerative colitis is featured by excessive intestinal inflammation and disrupted mucosal immune homeostasis. Codonopsis pilosula is the sovereign herb in Shenling Baizhu Formula for ulcerative colitis therapy, and Codonopsis pilosula polysaccharides are its key active components, while their immunomodulatory effects and molecular mechanisms in Shenling Baizhu Formula against ulcerative colitis remain elusive. This study explored the critical role of Codonopsis pilosula polysaccharides and the spermidine-AMPK-mediated mitophagy-cGAS-STING inflammatory axis in regulating intestinal inflammation and ameliorating ulcerative colitis. Methods A dextran sulfate sodium-induced ulcerative colitis mouse model was established to evaluate the indispensable role of Codonopsis pilosula polysaccharides in Shenling Baizhu Formula. Ultra-high performance liquid chromatography metabolomics and 16S ribosomal RNA sequencing were performed to identify key metabolites and gut microbiota. In vitro dextran sulfate sodium-induced ulcerative colitis cellular models (human colorectal adenocarcinoma Caco-2 and human normal colonic epithelial NCM460 cells) were constructed to validate the regulatory mechanism of spermidine on AMPK-mediated mitophagy, inflammatory balance and intestinal epithelial injury. Results Codonopsis pilosula polysaccharides were essential for the anti-ulcerative colitis efficacy of Shenling Baizhu Formula: removal of Codonopsis pilosula significantly aggravated colonic pathological lesions, intestinal inflammatory responses and oxidative stress in ulcerative colitis mice, whereas Codonopsis pilosula polysaccharides supplementation reversed these abnormal phenotypes ( P < 0.05). Metabolomics and microbiomics confirmed that Codonopsis pilosula polysaccharides exerted therapeutic effects via spermidine (a key metabolite in glutamate metabolism pathway) and Muribaculaceae. Critically, in vitro dual-cell experiments verified that spermidine activated AMPK-mediated mitophagy to directly inhibit the hyperactivation of cGAS-STING inflammatory axis, thereby restoring intestinal inflammatory homeostasis and alleviating epithelial cell injury in ulcerative colitis. Conclusion Codonopsis pilosula polysaccharides extracted from the sovereign herb Codonopsis pilosula in Shenling Baizhu Formula ameliorate dextran sulfate sodium-induced ulcerative colitis, and the core mechanism depends on spermidine-triggered AMPK-mediated mitophagy to suppress the cGAS-STING inflammatory signaling axis, which plays a pivotal role in regulating intestinal immunity and repairing mucosal epithelial damage.
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