Objective To prepare sinomenine nanoemulsion (Sin-NE) co-stabilized by Bletilla striata polysaccharide and glycyrrhizic acid, systematically characterize its properties, investigate its stability and drug release behavior, and further evaluate its anti-inflammatory effects through in vitro experiments. Methods The Sin-NE was prepared using the ultrasonic emulsification method. Its morphology, particle size, polydispersity index (PDI), and ζ potential were observed and determined. Optimal formulation parameters were screened via single-factor and orthogonal experiments, followed by formulation validation. pH value, turbidity, viscosity, encapsulation efficiency, and drug loading capacity were measured. Stability was investigated under centrifugation, storage, dilution, and heating conditions was evaluated, and in vitro drug release behavior was investigated. A RAW264.7 cell inflammation model was established to validate the effects of the Sin-NE on nitric oxide (NO), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α). Results The optimized formulation comprised 3% Coicis Semen oil, 0.5% combined emulsifiers, a 1∶9 mass ratio of Bletilla striata polysaccharide to glycyrrhizic acid, 10 min of ultrasonic treatment, and 325 W ultrasonic power. The resulting Sin-NE exhibited an average particle size of (145.30 ±2.35) nm, ζ potential of (-39.51 ±0.43) mV, PDI of 0.095 ±0.022, and pH value of 3.990 ±0.033. Encapsulation efficiency and drug loading capacity reached 88.78% and 26.14 mg/g, respectively. The nanoemulsion demonstrated excellent stability under centrifugation, storage, dilution, and heating conditions and displayed sustained-release characteristics. In vitro, it significantly inhibited NO release and effectively reduced IL-6 and TNF-α levels in inflamed cells. Conclusion The Sin-NE significantly enhances sinomenine’s bioavailability and anti-inflammatory activity, providing experimental evidence for developing novel traditional Chinese medicine nanoemulsion formulations.

Objective To prepare liposomes co-loaded with shikonin and ligustilide (Lip@Shi/Lig), optimize its preparation process, and to investigate its characterization and transdermal performance in vitro. Methods The CCK-8 assay and Synergy Finder analysis tool were used to determine the optimal combination ratio of shikonin and ligustilide based on cell viability. Lip@Shi/Lig was prepared using the thin-film dispersion method, and the optimal formulation of Lip@Shi/Lig was screened through single-factor investigation using encapsulation efficiency as the evaluation criterion. The quality evaluation of Lip@Shi/Lig was conducted using characterization methods, including appearance and morphology observation, particle size, polydispersity index (PDI), ζ potential determination, X-ray diffraction (XRD) analysis, and Fourier transform infrared spectroscopy (FT-IR) analysis. Finally, the transdermal permeability and dermal retention performance of Lip@Shi/Lig were investigated using the Franz diffusion cell method. Results The optimal combined ratio of shikonin and ligustilide was 1∶1. The optimal conditions were determined as follows: egg yolk lecithin concentration of 10 mg/mL, phospholipid-cholesterol ratio of 4∶1, total drug-phospholipid ratio of 1∶15, and ultrasonication time of 5 min. The resulting Lip@Shi/Lig exhibited regular, spherical vesicle morphology, with encapsulation efficiencies of shikonin and ligustilide at (98.16 ±0.67)% and (97.20 ±0.76)%, respectively, particle size of (88.62 ±0.26) nm, PDI of 0.246 ±0.013, and ζ potential of (-36.57 ±1.65) mV. XRD and FT-IR results indicated that shikonin and ligustilide were successfully encapsulated in the liposomes. The cumulative penetration of shikonin and ligustilide in Lip@Shi/Lig within 30 h were (82.97 ±0.72) μg/cm² and (81.57 ±3.59) μg/cm², respectively, with dermal retention rates of (6.12 ±0.18) μg/cm² and (8.08 ±0.04) μg/cm², respectively, both of which were significantly higher than those of the single drug and the single-drug-loaded liposomes. Conclusion Lip@Shi/Lig was successfully prepared with favorable transdermal properties and stability. It significantly enhanced the transdermal penetration and dermal retention of both shikonin and ligustilide, providing a solid experimental foundation for further in vivo studies and potential clinical applications.

Objective To explore the effect and mechanism of Yinyanghuo (Epimedii Folium) and Nüzhenzi (Ligustri Lucidi Fructus) on Buguzhi (Psoraleae Fructus)-induced liver injury in a rat model of kidney-yin deficiency based on the theory of “heterogeneous medicinals mutual restriction”. Methods The liver injury rats with kidney-yin deficiency model induced by Psoraleae Fructus was taken as a reference, Epimedii Folium and Ligustri Lucidi Fructus were respectively given for intervention, the changes in indicators of kidney-yin deficiency were evaluated by through body weight, anal temperature, fecal water content and cyclic adenosine monophosphate (cAMP)/cyclic guanosine monophosphate (cGMP) ratio in serum. The changes in liver function were evaluated by measuring the activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum, as well as by staining liver tissue with hematoxylin eosin (HE). Metabolomics, association analysis, ROC analysis and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis were combined to elucidate potential mechanisms. Results Epimedii Folium significantly decreased body weight and fecal water content in model rats (P < 0.05, 0.01), significantly increased anal temperature, cAMP/cGMP ratio and activities of ALT, AST (P < 0.05, 0.001), aggravated the infiltration of inflammatory cells in liver and exacerbated liver damage. Ligustri Lucidi Fructus significantly reversed the above changes. Metabolomics identified 17 common metabolites that were differentially regulated between Epimedii Folium and Ligustri Lucidi Fructus. Association analysis and ROC analysis further revealed six differential metabolites with good diagnostic efficacy, including ceramide and taurocholic acid, mainly involving eight pathways such as linoleic acid metabolism and sphingolipid metabolism. In addition, Epimedii Folium could significantly increase the levels of inflammatory factors such as interleukin-1β (IL-1β), IL-6, tumor necrosis factor-α (TNF-α) in serum of model rats (P < 0.05), and decrease anti-inflammatory factor IL-10 level. Ligustri Lucidi Fructus reversed the above changes. Conclusion Epimedii Folium and Ligustri Lucidi Fructus may affect the levels of inflammatory cytokines by differentially regulating liver linoleic acid metabolism, leading to differences in toxicity/efficacy in liver of rats with kidney-yin deficiency induced by Psoraleae Fructus. This study provides experimental basis for the theory of “heterogeneous medicinals mutual restriction” in traditional Chinese medicine, and provides reference for the research on reducing toxicity and increasing efficiency of Psoraleae Fructus.

Objective To investigate the effect of dandelion polysaccharide (DP) on lipid metabolism disorder in liver and gut microbiota structure in high-fat diet-induced obese mice. Methods A total of 40 SPF male C57BL/6J mice were randomly divided into control group, model group, orlistat (10 mg/kg) group, DP low-, high-dose (200, 400 mg/kg) groups, with eight mice in each group. Mice were fed with high-fat diet to establish an obesity model, and the model was simultaneously administered with ig for eight weeks. After the end of administration, mice were euthanized by removing their eyeballs and collecting blood. Serum was collected to measure the levels of total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C); Hematoxylin-eosin (HE) staining and Oil red O staining were used to observe the pathological changes of liver and epididymal fat in mice; qRT-PCR was used to detect the expressions of lipid metabolism related genes [carnitine palmitoyltransferase 1 (CPT-1), cytochrome P450 family 7 subfamily A member 1 (CYP7A1), peroxisome proliferator-activated receptor α (PPARα), fatty acid synthase (FAS), peroxisome proliferator-activated receptor γ (PPARγ), stearoyl-CoA desaturase 1 (SCD-1), hormone-sensitive lipase (HSL), adipose triglyceride lipase (ATGL)] in liver tissue; Feces were collected from each group of mice for microbiome sequencing. Results Compared with control group, the body weight, epididymal fat index, and levels of TC, TG, LDL-C in serum of mice in model group were significantly increased (P < 0.01, 0.001), HDL-C level were significantly decreased (P < 0.01). The structure of liver cells was damaged, the intercellular space was unclear, the arrangement of hepatic cords was significantly disordered, the area of vacuoles and the number of lipid droplets were significantly increased (P < 0.001), and the volume of epididymal fat cells was significantly increased with varying cell shapes and sizes. The expression levels of fat generation related genes FAS, PPARγ and SCD-1 mRNA were significantly increased (P < 0.01), while the expression levels of fat breakdown related genes HSL and ATGL mRNA were significantly decreased (P < 0.01). Compared with model group, DP low-, high-dose groups and orlistat group all reduced the body weight and epididymal fat index of obese mice to varying degrees, and improved the histopathological morphology of liver tissue and epididymal fat. The expression levels of fat synthesis related genes FAS, PPARγ and SCD-1 were significantly reduced in DP high dose group (P < 0.01), while the expression levels of fat breakdown and oxidation related genes HSL, ATGL, CTP-1, CYP7A1, PPARα mRNA were significantly increased (P < 0.01). Meanwhile, DP regulated the species composition of gut microbiota in obese mice to a certain extent, significantly increased the relative abundance of Blautia, s_unclassified_g_Lachnospiraceae_NK4A136_group, Lachnospiraceae_bacterium_28-4 and Helicobacter rodentium, decreased the relative abundance of Erysipelatoclostridium and Ruminococcus_torques_group, and their dominant strains were positively correlated with fat oxidation and decomposition genes HSL, ATGL, CTP-1, CYP7A1 and PPARα, and negatively correlated with fat synthesis genes FAS, PPARγ and SCD-1. Conclusion DP could significantly reduce the body weight of obese mice induced by a high-fat diet. The mechanism may be related to regulating beneficial bacteria to promote fat oxidation and decomposition and inhibit fat synthesis.

Objective To explore the mechanism by which Astragalus polysaccharides (APS) improves the consumption of adipose tissue in cancer cachexia (CC) using lipidomics and transcriptomics. Methods A random number table was used to divide 50 male mice into control group, model group, APS low-, medium-and high-dose (200, 400, 800 mg/kg) groups, with 10 mice in each group. CC model was constructed using abdominal sc CT-26 colorectal cancer cells. The treatment group was given 0.2 mL APS solution by ig daily, while the control group and model group were given an equal volume of physiological saline by ig for four consecutive weeks. The diameter of epididymal adipocytes was evaluated using hematoxylin-eosin (HE) staining; ELISA method was used to detect the levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), cyclic adenosine monophosphate (cAMP) in epididymal adipose tissue and level of free fatty acids (FFA) in plasma. After effectiveness and significance verification, transcriptome and lipidomics sequencing were performed on control group, model group and APS high-dose group to screen differentially expressed genes (DEGs) and differentially expressed lipids (DELs), and enrichment analysis was performed; Western blotting was used to verify the expressions of relevant proteins. Results High-dose APS significantly alleviated inflammatory infiltration in epididymal adipose tissue and reversed adipocyte atrophy (P < 0.01), significantly reduced the levels of TNF-α, IL-6, cAMP in adipose tissue and level of FFA in plasma (P < 0.01, 0.001), indicating that high-dose APS inhibits inflammation and lipolysis reactions in adipocytes. The lipidomics results showed that high-dose APS intervention reduced the expression levels of 66 DELs, including ceramides and their modifications, triacylglycerols, etc. Metabolic pathway enrichment analysis showed that these DELs mainly involved glycerophospholipid metabolism and sphingolipid metabolism pathways. The transcriptomic results showed that the phosphatidylinositol 3-kinase (PI3K)-protein kinase B (Akt) pathway was repeatedly enriched in DEGs, indicating that this pathway may be the core pathway for APS to function. In addition, the expressions of 18 DEGs was significantly down-regulated by APS, and the gene interaction network diagram showed that these genes had complex interactions with Akt. Western blotting results confirmed that APS intervention reversed the expression levels of Tribbles homologous protein 3 (TRIB3), phosphorylated Akt (p-Akt), phosphodiesterase 3B (PDE3B), phosphorylated protein kinase A (p-PKA), phosphorylated hormone sensitive lipase (p-HSL) and adipose triglyceride lipase (ATGL) (P < 0.05, 0.01, 0.001). Conclusion APS may alleviate inflammation in adipose tissue, reduce the synthesis of sphingolipids such as ceramides, regulate TRIB3-Akt axis, restore insulin resistance to lipolysis, promote lipid signaling, and improve the consumption of CC adipose tissue.

Objective To investigate the effect of Dendrobium officinale polysaccharide (DOP) on prediabetes (PDM) and explore its mechanism in improving PDMs through the gut short-chain fatty acids (SCFAs)-G protein-coupled receptor 43 (GPR43)-glucagon-like peptide 1 (GLP1) pathway. Methods SD Rats were randomly divided into control group, model group and DOP low-, high-dose (0.1, 0.2 g/kg) groups, with 10 rats in each group. Except for the control group, the remaining rats were fed a high-sugar and high-fat diet for six weeks to establish a PDM model. During the modeling period, the corresponding drugs were administered intravenously, and the control group and model group were given distilled water of equal volume. Changes in general physiological indicators, glucose/lipid levels, glucose regulation function, intestinal/pancreatic histopathology, fecal SCFAs contents, expressions of GPR43 and GLP1 and its receptor in intestinal/pancreatic were analyzed. Results Compared with model group, DOP significantly increased tail microcirculatory blood flow in PDM rats (P < 0.01) and decreased anal temperature (P < 0.01), significantly reduced levels of total cholesterol and low-density lipoprotein cholesterol in serum (P < 0.05, 0.01), significantly decreased the oral glucose tolerance test results, fasting blood glucose, 2 h postprandial glucose, homeostasis model assessment-insulin resistance index, and C-peptide levels in PDM rats (P < 0.05, 0.01), increased the insulin sensitivity index (P < 0.05), and alleviated the pathological damage of pancreas, ileum and colon. Gas chromatography results showed that DOP administration could significantly increase the levels of acetic acid, butyric acid, isovaleric acid and SCFAs in feces (P < 0.05, 0.01). ELISA, Western blotting and qRT-PCR results showed that DOP administration could significantly increase the plasma GLP1 level, as well as the mRNA and protein expressions of GPR43 and GLP1 receptor in ileum and pancreas (P < 0.05, 0.01). Conclusion DOP could improve PDM, and its effect may be related to enhancing gut-derived SCFA production, up-regulating GPR43 expressions in intestine and pancreas, subsequent promoting GLP1 secretion, restoring β-cell function and improving glucose regulation.

Objective To investigate the effect and mechanism of notoginsenoside R₁ (NGR1) on cardiac remodeling after myocardial ischemia-reperfusion injury (MIRI) in mice. Methods A model of cardiac remodeling after myocardial ischemia-reperfusion injury (MIRI) was established by ligating the anterior descending branch of the coronary artery in mice. After three weeks of intervention with NGR1, the cardiac index and heart weight/tibia length of mice were measured. The left ventricular end-diastolic volume (LVEDV), left ventricular end-systolic volume (LVESV), left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) were measured using a small animal ultrasound instrument. The cross-sectional area of myocardial cells was calculated by wheat germ agglutinin (WGA) fluorescence staining. The collagen volume fraction (CVF) of heart was calculated by Masson staining. The protein expressions of collagen I, collagen III, atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), transferrin receptor 1 (TFR1), acyl-CoA synthetase long-chain family member 4 (ACSL4), solute carrier family 7 member 11 (SLC7A11) and glutathione peroxidase 4 (GPX4) in myocardial tissue were detected by Western blotting. The possible mechanism of NGR1 inhibiting cardiac remodeling after MIRI was preliminarily explored through network pharmacology. The contents of iron ions (Fe²⁺) and lipid peroxide (LPO) in myocardial tissue were detected using kits. The damage of myocardial mitochondria was observed by transmission electron microscopy. Results Compared with sham group, cardiac index, heart weight/tibia length, LVEDV, LVESV and CVF of mice in model group were significantly increased (P < 0.05, 0.01), protein expressions of collagen I, collagen III, ANP, BNP, TFR1 and ACSL4 in myocardial tissue were significantly increased (P < 0.01), contents of Fe²⁺ and LPO were significantly increased (P < 0.05, 0.01), mitochondrial membrane density was increased and cristae was decreased, SLC7A11 and GPX4 protein expressions were significantly decreased (P < 0.01). Compared with model group, cardiac index, heart weight/tibia length, LVEDV, LVESV and CVF of mice in NGR1 administration groups were significantly decreased (P < 0.05, 0.01), protein expressions of collagen I, collagen III, ANP, BNP, TFR1 and ACSL4 were significantly decreased (P < 0.05, 0.01), contents of Fe²⁺ and LPO were significantly decreased (P < 0.05, 0.01), mitochondrial damage was improved, SLC7A11 and GPX4 protein expressions were significantly increased (P < 0.05, 0.01). Conclusion NGR1 could significantly improve cardiac remodeling in mice after MIRI, and its mechanism may be related to the activation of SLC7A11/GPX4 pathway to inhibit ferroptosis of cardiomyocytes.

Objective To investigate the subcellular distribution patterns of the combination of cantharidin (CTD) and baicalin (BA) based on cell pharmacokinetics and elucidate the synergistic mechanism against hepatocellular carcinoma. Methods Using human hepatoma HepG2 cells as the model, the dynamic concentrations of the drugs in whole cells and organelles (nucleus, mitochondria, endoplasmic reticulum, and lysosomes) over 12 h after administration of single drugs (CTD 6 μg/mL, BA 30 μg/mL) and their combination (CTD 6 μg/mL + BA 30 μg/mL) were quantitatively analyzed by ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Pharmacokinetic parameters were calculated using a non-compartmental model with Phoenix WinNonlin software. Results At the whole-cell level, the combination increased the intracellular area under the curve (AUC_0—t) of CTD by 48.9% and decreased its clearance (P < 0.05), but did not significantly affect the pharmacokinetics of BA. At the subcellular level, the combination significantly increased the AUC_0—t of CTD and BA in nucleus, lysosome, mitochondria and endoplasmic reticulum by 93.5%, 46.4%, 38.3%, 52.3%, and 68.4%, 40.0%, 41.0%, and 46.7%, respectively (P < 0.05, 0.01). Furthermore, the time to reach peak concentration (t_max) for both drugs in mitochondria was earlier in the combination group, and the mean residence time (MRT_0—t) of CTD in the endoplasmic reticulum was significantly prolonged (P < 0.01), indicating a spatiotemporally synergistic drug delivery optimization. Conclusions The combination of CTD and BA could synergistically optimize the distribution of drugs in key subcellular structures such as nucleus, mitochondria, endoplasmic reticulum and lysosomes, and may enhance the anti-hepatocellular carcinoma effect by inducing DNA damage, accelerating mitochondria-mediated apoptosis and promoting endoplasmic reticulum stress, which provides a theoretical basis for the design of traditional Chinese medicine combinations based on organelle targeting.

Objective To explore “state and target” pathological mechanism of sepsis by comprehensively analyzing the single-cell transcriptomics and transcriptomic datasets related to sepsis, construct the prognostic model for “state and target” of sepsis, and explore potential target Chinese medicines and active ingredients. Methods Differential expression gene analysis and venn diagram analysis were used to identify “state and target” of sepsis related genes. Key module genes were obtained using the random walk algorithm through the connectivity of the protein-protein interaction (PPI) network. Gene ontology (GO) biological process (BP) and Kyoto encyclopedia of genes and genomes (KEGG) were used to analyze the biological functions of key module genes. To explore the role of “state and target” of sepsis related genes by single-cell transcriptome analysis. A prognostic model for “state and target” of sepsis was constructed using gene expression data and clinical survival data from sepsis patients. The expression levels of prognostic genes for “state and target” of sepsis were validated by constructing sepsis mouse model and real-time quantitative polymerase chain reaction (RT-PCR). Through reverse network pharmacology, potential targeted traditional Chinese medicines and their active components were screened, and molecular docking was used to verify the binding performance between active ingredients and prognostic targets. Results A total of 70 genes related to “toxic state” of sepsis, 67 genes related to “stasis state” of sepsis and 54 genes related to “deficiency state” of sepsis were obtained. The key module genes of “toxic state” of sepsis were closely related to inflammation, the key module genes of “stasis state” of sepsis were closely related to coagulation, and the key module genes of “deficiency state” of sepsis were closely related to cell homeostasis. AUCell score showed that monocyte had the highest score for “toxic state” and “deficiency state” of sepsis. Platelet had the highest score for “stasis state” of sepsis. The activities of metabolic and inflammatory related genes and pathways were significantly up-regulated in monocytes with high “toxic state” score. Platelet activation and coagulation/thrombosis related genes and pathways were significantly up-regulated in platelet with high “stasis state” score. Compared with control group, there were a large number of down-regulated genes in monocytes of sepsis patients, which were involved in antigen presentation, cell energy metabolism, cell cycle, protein synthesis, etc. The prognostic model for “state and target” of sepsis constructed based on eight targets [interleukin-1 receptor type 2 (IL1R2), ADP ribosylation factor like GTPase 4C (ARL4C), cytochrome C oxidase subunit 7B (COX7B), eukaryotic translation initiation factor 2 subunit gamma (EIF2S3), immediate early response 3 (IER3), LSM1 homolog (LSM1), paired basic amino acid cleaving enzyme (FURIN) and ankyrin repeat domain-containing protein 9 (ANKRD9)] had good prognostic predictive ability, and the expression levels of these genes were validated using animal experiments and RT-PCR. Based on the bioinformatics analysis tool for molecular mechanism of traditional Chinese medicine (BATMAN-TCM) database and database of constituents absorbed into the blood and metabolites of traditional Chinese medicine (DCABM-TCM), reverse network pharmacology analysis had screened multiple Chinese herbal medicines with effects of clearing heat and detoxifying, activating blood and resolving stasis, and reinforcing healthy qi and their active ingredients entering the blood. Molecular docking showed that representative active ingredients have good binding properties with prognostic targets. Conclusion Activation of inflammatory response driven by monocyte metabolic reprogramming may be the main pathological mechanism of “toxic state” of sepsis. Platelet activation-coagulation/thrombosis may be the main pathological mechanism of “stasis state” of sepsis. Monocyte dysfunction-immunosuppression may be the main pathological mechanism of “deficiency state” of sepsis. A model based on eight genes can serve as a risk prediction model for the prognosis of “state and target” of sepsis.

Objective To explore the “medicine-efficacy” associations of traditional Chinese medicine (TCM) in the treatment of sepsis, identify core herbsrelated to reducing mortality and improving key clinical outcomes such as the acute physiology and chronic health evaluation II (APACHE II) score, and provide evidence-based references for TCM-assisted treatment of sepsis. Methods A systematic search of Chinese and English databases was conducted up to September 2024 to collect clinical studies on TCM compound prescriptions for sepsis. A comprehensive sepsis TCM formula database was established. Nine machine learning algorithms were compared using ten-fold cross-validation, and the optimal model for each clinical outcome was interpreted through the Shapley additive explanations (SHAP) method to identify key herbs and their contribution directions. Results The multilayer perceptron showed the best performance in predicting TCM syndrome scores, overall effectiveness, inflammatory and immune indicators, biochemical parameters, organ dysfunction scores, and mortality; logistic regression performed best for blood gas analysis along with gastrointestinal function and intestinal mucosal barrier outcomes; and the support vector machine achieved optimal predictive performance for routine blood tests and APACHE II scores. SHAP analysis revealed that Dihuang (Rehmanniae Radix), Zhishi (Aurantii Fructus Immaturus), Huangqi (Astragali Radix), Fuzi (Aconiti Lateralis Radix Praeparata), Huangqin (Scutellariae Radix), and Houpo (Magnoliae Officinalis Cortex) had positive contributions across outcomes such as mortality, APACHE II score, inflammatory markers, and gastrointestinal function, forming the core nodes of the “medicine-efficacy” network. Conclusion This study established a clinically oriented “medicine-efficacy” association network through multi-model comparison and explainable machine learning analysis. The findings highlight the potential key roles of several core herbs in improving major clinical outcomes of sepsis, providing data support and evidence-based basis for precise syndrome differentiation and medication in TCM, as well as for the research and development of new TCMs.