Liver injury is a clinical disorder caused by toxins, drugs, and alcohol stimulation without effective therapeutic approaches thus far. Scutellarin (SCU), isolated from the edible herb Erigeron breviscapus (Vant.) Hand. -Mazz. showed potential hepatoprotective effects, but the mechanisms remain unknown. In this study, transcriptomics combined with nontargeted metabolomics and 16S rRNA amplicon sequencing were performed to elucidate the functional mechanisms of SCU in carbon tetrachloride (CCl₄)–induced liver injury in mice. The results showed that SCU exerted potential hepatoprotective effects against CCl₄-induced liver injury by repressing CYP2E1 and IκBα/NF-κB signaling pathways, modulating the gut microbiota (especially enriching Lactobacillus), and regulating the endogenous metabolites involved in lipid metabolism and bile acid homeostasis. SCU originates from a functional food that appears to be a promising agent to guard against liver injury.

Purpose: Rodgersia sambucifolia Hemsl (also known as Yantuo) is a traditional Chinese medicine commonly utilized as a medicinal herb with its rhizomes, mainly used to regulate the immune function of the human body. However, relatively few studies have investigated its active components and potential mechanisms of action in vivo. Methods: First, the chemical composition in vitro was identified and analyzed using the UPLC-Q-TOF MS/MS technique. Cyclophosphamide (CTX) was then administered intraperitoneally to rats to establish an immunosuppression model. Physiological and biochemical parameters, organ indices, and histopathological findings were evaluated for efficacy. Subsequently, potential biomarkers in rat serum were identified using multivariate statistical analysis and enriched and topologized using online platforms such as MetaboAnalyst and KEGG to reveal the critical metabolic pathways and their roles in the immunomodulatory network. Finally, the integrated analysis of components in vivo and in vitro, along with metabolic pathways, was performed using network pharmacology and molecular docking technology to elucidate the mechanisms of their roles in organismal immunity. Results: A total of 28 chemical components in vitro were identified, while pharmacodynamic experiments confirmed the immunomodulatory effects of Yantuo, especially in the high-dose administration group. Metabolomics analysis showed that 37 potential immune-related biomarkers were identified in positive and negative ion modes, involving 16 metabolic pathways such as arginine biosynthesis, pyrimidine metabolism, and riboflavin metabolism. The results of network pharmacology and molecular docking indicated that Yantuo may affect 7-O-galloyl-catechin, Cynaroside, Quercetin-7-O-beta-D-glucopyranoside, and 1.6-bis-O-galloyl-beta-D-glucose through interactions with the immune system, with significant pathways of action including galactose metabolism, glycolysis/gluconeogenesis, pyrimidine metabolism, and riboflavin metabolism. Conclusion: In our experiments, we confirmed the organismal modulatory effect of Yantuo on immunocompromised rats, clarified the key components, target proteins, and pathways of its possible action, and provided possibilities for follow-up studies.

Background: Periplaneta americana L . is one of the most famous traditional Chinese medicines (TCMs) , and CII-3 is the major bioactive extract of Periplaneta americana L. In recent years, CII-3 has gradually attracted the attention of researchers for its powerful capacity for treating immunocompromised diseases. However, systematical chemical composition investigation and mechanisms on immunomodulation of CII-3 have not been thoroughly scrutinized. Methods: The chemical ingredients of CII-3 were determined by ultra-performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS), and immune-enhancing mechanisms and active constituents were investigated by integration of metabolomics with network pharmacology and molecular docking. Results: A total of 25 components were identified in the aqueous extract of CII-3, among which 10 were unambiguously confirmed by comparison with reference standards. In immunosuppressed rats, oral administration of CII-3 normalized biochemical profiles by promoting the secretion of immune-related cytokines (IL-2, IL-6) and immunoglobulins (IgG, IgM), significantly improving immune organ indices, and alleviating pathological lesions in the thymus and spleen. Furthermore, CII-3 stimulation attenuated CTX-induced upregulation of IL-6 mRNA levels and concurrently enhanced IL-2 mRNA expression in these two immune organs. Based on metabolomics analysis, 27 differential metabolites along with 7 crucial metabolic pathways including starch and sucrose metabolism, arginine biosynthesis, porphyrin and chlorophyll metabolism, nicotinate and nicotinamide metabolism, steroid biosynthesis, pyrimidine metabolism and tryptophan metabolism were regulated after CII-3 treatment, suggesting that CII-3 has the potential to target these metabolites and key pathways to improve immunosuppression. Nitric oxide synthase 1 (NOS1), nitric oxide synthase 3 (NOS3), acetylcholinesterase (ACHE), cluster of differentiation 38 (CD38) and poly (ADP-ribose) polymerase 1 (PARP1) were considered as the pivotal targets, and ginsenine, cyclo (Tyr-Asp), guanosine, tryptophan and inosine were deemed potential active components of CII-3 in alleviating immunosuppression. Conclusion: The proposed approach provides a valuable evidence for demonstrating the material basis of CII-3. The results obtained in the present study elucidated that the potential immunomodulatory activity of CII-3 might be closely associated with these crucial targets, which can modulate the levels of major metabolites through multiple metabolic pathways, thus ameliorating immunosuppression.

Background: Asthma, a chronic inflammatory respiratory disease with significant global health burden, faces limitations in current therapies, necessitating novel therapeutic strategies. The plant Geranium wilfordii Maxim. (GWM), a traditional Chinese herbal medicine with diverse pharmacological activities and clinical applications, has been traditionally used in the treatment of rheumatism, numbness, infectious diseases, dermatosis, tumors and other disease by the Bai, Miao, Yi minority people of Southwest China for generations. Earlier research in our lab also demonstrated that GWM exhibits anti-asthmatic activity, but the mechanism of action remains unclear. Aim: To investigate the anti-asthmatic mechanisms of GWM by identifying compounds and elucidating key molecular targets involved in immune cell regulation through integrated computational and experimental approaches. Methods: We employed UPLC-QE-Orbitrap-MS to identify active compounds. Network pharmacology and machine learning analyses were conducted to identify key hub genes, followed by validation through Mendelian randomization analysis, molecular docking, and animal models. Immune infiltration and single-cell RNA sequencing analyses using publicly available Gene Expression Omnibus (GEO) datasets, combined with mediation Mendelian randomization (MR), were performed to elucidate the underlying cellular mechanisms. Results: A total of 43 compounds were identified in GWM. Network pharmacology and machine learning prioritized NOTCH2, HDAC2, and MAPK1 as hub targets, validated using MR and molecular docking. Subsequent in vivo experiments validated the regulatory effects of GWM on the expression levels of these hub genes and demonstrated its therapeutic efficacy in asthma. Further analysis showed that GWM regulation of these hub genes may subsequently affect the activity of CD4 ⁺ T cells and regulatory T cells, potentially contributing to its therapeutic effects against asthma. Conclusion: This study provides novel evidence for the potential therapeutic activity of GWM in asthma. By targeting key hub genes such as NOTCH2, HDAC2, and MAPK1, GWM may modulate immune cell activity, thereby contributing to its anti-asthmatic effects.

Objectives: A LC-MS/MS method for the detection of ulipristal acetate in humans which was not only simple and rapid in sample pretreatment process and chromatographic condition, but also highly selective and sensitive in MS condition was developed, and the fully validated method was applied for investigating the pharmacokinetic properties of ulipristal acetate following single oral administration of Esmya in healthy Chinese subjects for the first time. Materials and methods: After single-step protein precipitation with methanol, ulipristal acetate and ulipristal acetate-d3 (IS) were chromatographed on an ACE Excel 3 C-18-PFP column with gradient elution procedure, and detected by positive electrospray ionization with multiple reaction monitoring mode using the respective precursor-product ion transitions of m/z 476.2 -> 134.1 for ulipristal acetate and m/z 479.3 -> 416.2 for IS. Results: The assay has desirable accuracy and precision over a wide linear range of 0.0500 - 100 ng/mL for ulipristal acetate, and no interference caused by endogenous compounds was detected. It also exhibits satisfactory matrix effect, extraction recovery, and stability. C-max of ulipristal acetate after oral administration of a single 5-mg dose of 47.7 +/- 27.7 ng/mL was detected at 0.91 +/- 0.98 h (t(max)) with AUC(0-t) of 112 +/- 49 ng.h/mL. Ulipristal acetate was eliminated slowly with T-1/2 of 46.4 +/- 14.0 h. Conclusions: The major advantages of the current approach involve one-step precipitation of plasma protein with methanol, high selectivity with a lower limit of quantitation of 0.0500 ng/mL, small plasma volume of 50 mu L for processing and a short run time of 4 min per sample, which allow its applicability for a pharmacokinetic study in healthy Chinese subjects.