Glioblastoma (GBM) is an aggressive primary brain tumor that, partly due to its hypoxic tumor microenvironment (TME), is extremely difficult to treat. In our study, RNA sequencing and quantitative reverse-transcription PCR (qRT-PCR) analysis identified differential expression of chromobox 6 (CBX6) and carbonic anhydrase 9 (CA9) in GBM cells under hypoxic conditions. Downregulation of CBX6, a member of the Polycomb group proteins, occurs under hypoxic conditions and is associated with higher-grade tumors and worse patient survival. Here, we show that silencing CBX6 in GBM cells promotes proliferation, migration, and invasion, while its overexpression yields the opposite effects, indicating its role as a negative regulator of tumor aggressiveness. CA9, upregulated by hypoxia, contributes to an acidic environment supporting tumor growth, a more aggressive GBM phenotype, and treatment resistance. Our qRT-PCR and short hairpin RNA (shRNA)-mediated knockdown experiments demonstrate an inverse relationship between CBX6 and CA9 expression across various human and murine GBM cell lines. Chromatin immunoprecipitation (ChIP) assays with multiple primers confirmed that CBX6 binds to the CA9 promotor, suggesting that CBX6 regulates CA9 expression. Our findings highlight CBX6 and CA9 as potential therapeutic targets, offering insights into GBM biology and the response to hypoxia.

Liver fibrosis is a progressive pathological process triggered by chronic liver disorders, which may progress to hepatocellular carcinoma (HCC) if left untreated. Currently, there are no specific therapeutic agents for liver fibrosis, highlighting an urgent need for novel pharmacological strategies. Tomatidine (TD), a major steroidal glycoal-kaloid abundant in immature tomato fruits, leaves, and stems, exhibits diverse biological activities including anti-inflammation, anti-tumor effects, and autophagy regulation. However, its role in liver fibrosis and the underlying molecular mechanisms remain incompletely understood. In this study, we combined network pharmacology, molecular docking, and experimental validation to investigate the potential effects of TD against liver fibrosis and its associated mechanism of action. In vitro experiments using the human hepatic stellate cell (HSC) line LX-2 demonstrated that TD inhibited HSC proliferation in a dose- and time-dependent manner, and downregulated the expression of fibrosis-related markers α-smooth muscle actin (α-SMA) and collagen type I α1 chain (COL1A1) at the gene, protein, and cellular levels. Network pharmacology analysis identified 18 common targets between TD and liver fibrosis, with core targets including MAPK3, RELA, and MAPK1 involved in intracellular signal transduction and stress-activated MAPK cascade. Although no autophagy-related targets were identified in the current database among these common targets, pharmacological evidence and experimental validation confirmed that TD promoted autophagy in LX-2 cells, as indicated by reduced P62 expression, increased LC3-II/LC3-I ratio and Beclin-1 levels, and enhanced autophagic flux. Further mechanism exploration revealed that TD exerted its autophagy-promoting effect by regulating the ERK/MAPK-mTOR-ULK1 signaling pathway: TD suppressed the phosphorylation of ERK and mTOR, while activating ULK1 phosphorylation. Molecular docking verified stable binding affinity between TD and key proteins in this pathway (ERK, MAPK, mTOR, ULK1) as well as autophagy-related proteins (P62, Beclin-1, LC3) and fibrosis-related protein COL1A1, with specific amino acid residues mediating hydrogen bond formation. Collectively, our findings demonstrate that TD modulates fibrosis-related markers in hepatic stellate cells by promoting autophagy in HSCs via the ERK/MAPK-mTOR-ULK1 pathway. This study enriches the biological function research of TD and provides a novel potential candidate and theoretical basis for the development of anti-liver fibrosis therapeutics.

The fruits of Amomum maximum Roxb. (Zingiberaceae), traditionally used for gastrointestinal disorders, have been less phytochemically investigated compared to its rhizomes. In this study, four undescribed compounds (1-3,12) together with 15 known compounds were isolated from a 75% ethanol extract of A. maximum fruits, including eleven diarylheptanoids and eight labdane diterpenes. Their structures were unequivocally elucidated by spectroscopic methods, including NMR spectroscopy, computational NMR methods, and ECD. Compounds 1, 3-6, 10, and 11 significantly inhibited the release of NO, IL-6, and TNF-α in LPS/IFN-γ-stimulated RAW 264.7 macrophages. Furthermore, the integration of network pharmacology, molecular docking, and molecular dynamics simulations identified compounds 1, 3-6, 10, and 11 as promising candidates, demonstrating favorable binding affinity and stability with the active target AKT1. This study demonstrates that diarylheptanoids are the major anti-inflammatory components in A. maximum fruits, which not only validates the ethnopharmacological use of this herb, but also provides new anti-inflammatory agents derived from natural products.

A phytochemical investigation of the rhizomes of Zingiber montanum was undertaken, leading to the isolation and identification of eight previously undescribed compounds. These compounds were categorized as three phenylbutene dimers (1-3) and five saturated phenylbutene-type derivatives (4-8). Their structures were unequivocally determined through extensive spectroscopic analysis, including 1D/2D NMR, UV, IR and MS spectroscopy. The absolute configurations of these compounds were established by a combination of electronic circular dichroism calculations and DP4+ probability analysis. The anti-inflammatory activity of these compounds was evaluated on the LPS/IFN-γ-stimulated RAW 264.7 macrophages. The results revealed that compounds 1 and 4-8 exhibited significant inhibition on the production of nitric oxide at the concentration of 10 μM. These findings enhance the understanding of the chemical composition of Zingiber montanum, and provide a robust scientific foundation for its traditional application in the treatment of inflammatory disease, highlighting these novel compounds as promising leads for further anti-inflammatory agent development.

Rationale:Ubiquitin-specific peptidase 22 (USP22), a deubiquitinase and component of the "Death-from-Cancer" 11-gene signature, is overexpressed in multiple malignancies and linked to recurrence, therapy resistance, and poor prognosis. Its role in KRAS/p53-driven lung cancer and the response to immune checkpoint inhibitors (ICIs) remains poorly defined. Here, we investigated USP22 as a potential therapeutic target in KRAS/p53-driven lung cancer. Methods:A conditional Usp22 knockout (Usp22-KO) was generated in the KRASG12D; p53-/- (KP) mouse model. Cancer progression was monitored by micro-computed tomography (micro-CT). Multiplex immunofluorescence (mIF), RNA sequencing, and spatial transcriptomics profiled cancer and tumor microenvironment (TME) changes. Responses to anti-PD-1/PD-L1 therapies were compared between KP and Usp22-KO KP (KPU-) lung cancers. Results:USP22 was highly expressed in early-stage KRAS/p53-driven mouse lung cancers and strongly correlated with proliferation marker Ki67. Usp22 deletion suppressed cancer growth, prolonged survival, and promoted cancer differentiation. Spatial transcriptomics and mIF revealed reduced CD206+ M2 macrophages, myeloid-derived suppressor cells (MDSCs), TGF-β1, and angiogenesis, along with increased functional CD8+ T cells. Mechanistically, USP22 regulated gene expression and protein stability, reducing c-Myc, PD-L1, TGF-β1, and SPARC upon Usp22 loss. Compared with KP cancer, KPU- and SPARC-knockdown KP cancers showed reduced macrophage chemotaxis and impaired basal- and TGF-β1-induced M2 polarization of RAW264.7 cells, suggesting that TGF-β1 and SPARC downregulation partially contributes to decreased M2 macrophage infiltration in KPU- cancers. Notably, Usp22 loss enhanced the efficacy of anti-PD-L1 and anti-PD-1 therapies in orthotopic and subcutaneous KP lung cancer models, respectively. USP22 and SPARC expression were also strongly correlated in human lung cancers. Conclusions:USP22 promotes progression and immune evasion in KRAS/p53-driven lung cancer. Targeting USP22 reprograms the TME, suppresses oncogenic signaling, and sensitizes tumors to ICI, establishing USP22 as a promising therapeutic target.

ETHNOPHARMACOLOGICAL RELEVANCE:Danshensu (DSS) is one of the water-soluble components extractable from the traditional Chinese medicine Salvia miltiorrhiza Bge., exhibiting pharmacological effects such as promoting blood circulation, dilating coronary arteries, and improving cerebral blood flow. The Danshensu derivative (OZD-1) obtained through the derivatization of DSS is a potential multi-target drug for the central nervous system, however, its mechanism of action against cerebral ischemia-reperfusion injury (CIRI) remains unclear. AIM OF THE STUDY:Systematically investigating the therapeutic potential and mechanisms of action of Danshensu derivative against cerebral ischemia-reperfusion injury. MATERIALS AND METHODS:Rat brain microvascular endothelial cells (RBMVECs) were cultured in vitro to establish an oxygen-glucose deprivation/reoxygenation (OGD/R) injury model. Groups included a control group, an OGD/R model group, and OZD-1 low-dose (12.5 μmol/L), medium-dose (25 μmol/L), and high-dose (50 μmol/L) groups. Cell viability, migration capacity, and vascular lumen formation were assessed using the CCK-8 assay, cell scratch assay, and matrigel matrix gel assay, respectively. In vivo, a transient middle cerebral artery occlusion (tMCAO) model was established in rats. Animals were randomly divided into the sham, model, OZD-1 (35, 70, 140 mg/kg), Edaravone (Eda), and DSS groups. Daily oral administration was performed post-surgery for 14 consecutive days. Tissue pathology staining, behavioral tests, and regional cerebral blood flow imaging assessed brain tissue damage, cognitive function, and ischemic side cerebral blood flow recovery, respectively. Transcriptome sequencing analyzed differential gene expression and pathway enrichment patterns. Western blot detection measured expression levels of proteins related to the PI3K-AKT-CREB signaling pathway, phosphoproteins, downstream apoptosis-related proteins, and CD31, CD34, and VEGFA proteins. RESULTS:In vitro experiments demonstrated that OZD-1 dose-dependently enhanced the viability of RBMVECs following OGD/R injury, significantly improving cell migration and luminal formation capabilities. In vivo studies revealed that compared to the model group, rats in all OZD-1 dosage groups exhibited markedly improved cognitive function, significantly restored cerebral blood flow in the ischemic hemisphere, and substantially reduced pathological brain tissue damage. Transcriptome sequencing results indicated significant enrichment of genes associated with the PI3K-AKT signaling pathway following OZD-1 intervention. Western blot experiments confirmed that OZD-1 significantly upregulates the phosphorylation levels of proteins related to the PI3K-AKT-CREB signaling pathway in OGD/R-injured cells and brain tissue from tMCAO rats, thereby promoting VEGFA-mediated angiogenesis and inhibiting apoptosis. To further verify pathway involvement, in vitro inhibition experiments were performed in RBMVECs using the PI3K inhibitor LY294002 and CREB inhibitor 666-15. These inhibitors abolished the OZD-1-induced upregulation of p-PI3K, p-AKT, and p-CREB, and reversed its protective effects on cell viability, migration, and tube formation. These results confirm that OZD-1 protects vascular endothelial cells directly via activating the PI3K-AKT-CREB pathway. CONCLUSION:OZD-1 exhibits significant neuroprotective effects against CIRI in rats, improving cognitive function, promoting vascular regeneration in ischemic areas, repairing damaged RBMVECs, and reducing apoptosis. Its mechanism of action is associated with the activation of the PI3K-AKT-CREB-VEGFA signaling pathway.

De novo purine synthesis is required to maintain tumor growth; however, its impact on therapy resistance remains unclear. Here, through a dynamic BH3-priming-based CRISPR screen, we found that deletion of ADSS2, which encodes the adenylosuccinate synthase 2 enzyme essential for adenosine monophosphate (AMP) synthesis, re-sensitizes drug-resistant acute myeloid leukemia cells to venetoclax and a myeloid cell leukemia-1 (MCL1) inhibitor. Single-cell sequencing analysis of patient-derived xenograft samples revealed a positive association of high ADSS2 activity in TP53-mutant cells with poor responsiveness to venetoclax. We developed an ADSS2 antagonist, which synergized with BH3 mimetics to promote apoptosis in preclinical models. Mechanistically, sensitization mediated by ADSS2 targeting correlated with downregulated AMP-activated protein kinase activity, which in resistant cells promotes mitophagy to eliminate damaged mitochondria after BH3 mimetic treatment. These data show that AMP synthesis promotes BH3 mimetic resistance and that combining ADSS2 targeting with BH3 mimetics represents a promising anti-cancer approach.

Mulberry leaf, as a traditional Chinese medicinal plant, has been utilized in the treatment of various diseases, including diabetes, cardiovascular diseases, inflammatory disorders, and liver diseases. However, the mechanisms underlying its therapeutic effects on non-alcoholic fatty liver disease (NAFLD) remain unclear. Therefore, this study aims to investigate the potential mechanisms of mulberry leaf extract (MLE) in the treatment of NAFLD. The chemical composition of MLE was analyzed using ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS). The NAFLD mice model was induced by a high-fat, high-fructose, and high-cholesterol (HFFC) diet, followed by intervention with MLE. The results indicated that the administration of MLE notably reduced the obesity (p < 0.05), oxidative stress (p < 0.05), inflammation (p < 0.05), and ECM deposition (p < 0.05) induced by the HFFC diet, restored the parameters of liver function, and attenuated the pathological changes. Utilizing a combination of integrated liver non-targeted metabolomics, network pharmacology, and transcriptomic approaches, we deciphered the molecular mechanisms by which MLE exerted its therapeutic effects in the treatment of NAFLD. In detail, our findings revealed that MLE suppressed the TGFβ1/Smad3 and NF-κB signaling pathways to ameliorate fibrosis and inflammation. This study provided novel insights into the correlation between MLE and NAFLD progression, offering a scientific foundation for the prospective use of MLE in the treatment of NAFLD.

Seventeen compounds, including seven undescribed compounds named Zingiberenes A-G (1-7), were isolated from the traditional medicinal plant Zingiber montanum. Their planar structures and absolute configurations were established by comprehensive spectroscopic analysis and ECD calculations. Evaluation of anti-inflammatory activity in LPS/IFN-γ-stimulated RAW264.7 macrophages showed that compounds 1 and 4 significantly inhibited NO production (p < 0.01), with 4 exhibiting the strongest effect. At 10 µM, both compounds also suppressed the secretion of pro-inflammatory cytokines TNF-α and IL-6 by 30%-50% (p < 0.01). Network pharmacology analysis predicted multi-target anti-inflammatory mechanisms involving IL-6, GAPDH, and PTGS2. Molecular docking further supported these findings, demonstrating that compound 4 forms a stable complex with IL-6, which aligns with the experimental NO and cytokine data. This study offers valuable insights for future research on the anti-inflammatory mechanisms of phenylbutene derivatives.

Eczema, a chronic inflammatory skin condition, causes persistent discomfort and impairs patients' quality of life. Cnidium monnieri has been traditionally used for its anti-inflammatory and antipruritic properties. The present study aimed to develop an optimized formulation of Cnidium monnieri volatile oil cream (CMVOC) and evaluate its anti-inflammatory effects in an eczema model. A single-factor and response surface optimization approach was used to determine the optimal cream formulation. The formulation was evaluated based on appearance, physical stability, particle size and moisturizing capacity. An eczema model was induced using 2,4-dinitrochlorobenzene (DNCB) in mice. Serum levels of IL-6 and IL-17 were measured by ELISA, skin pathology was assessed by hematoxylin and eosin and toluidine blue staining and JAK2/STAT3 expression was detected by immunohistochemistry to determine the activation status of the related signaling pathway. The optimized formulation contained 4.45 g octadecanol, 4 g Vaseline, 2.2 g liquid paraffin, 0.81 g isopropyl myristate Estergel®, 1.6 g Cnidium monnieri volatile oil, 0.39 g sodium dodecyl sulfate, 1.16 g glycerol, 0.04 g nibergin ethyl ester and 25.35 g Distilled Water. In vivo, CMVOC markedly improved skin lesions in DNCB-induced mice, reduced serum IL-6 and IL-17 levels and alleviated epidermal thickening, edema and inflammatory infiltration. Immunohistochemistry further demonstrated suppressed activation of the JAK2-STAT3 signaling pathway. CMVOC effectively mitigated eczema-related inflammation by reducing pro-inflammatory cytokines and inhibiting JAK2-STAT3 pathway activation, providing experimental support for its potential as a topical therapy for eczema.

Background Rheumatoid arthritis (RA) is characterized by synovial hyperplasia and a parallel dysbiosis of the gut microbiota. Wedelolactone (WL), a natural bioactive compound, exhibits potent anti-inflammatory and immunomodulatory activities; however, its therapeutic potential in RA is unknown. Purpose This study aimed to evaluate the potential effect of WL on RA and explore its underlying mechanism. Methods The anti-arthritic activity of WL was assessed in wild-type and antibiotic‑treated (ABX) collagen‑induced arthritis (CIA) mice in vivo and in human rheumatoid-arthritis synovial fibroblasts (MH7A) cells and murine rheumatoid-arthritis fibroblast-like synoviocytes (RA-FLS) cells in vitro. Additionally, fecal microbiota transplantation (FMT), 16S rDNA sequencing, intestinal barrier integrity assays, splenic and colonic Th17/Treg analyses and targeted metabolomics of short-chain fatty acids (SCFAs) were performed to clarify its microbiota-directed actions. Finally, RNA-seq coupled with chemical inhibition and genetic knockdown with siRNA were employed to explore its effect on the hyperproliferation of synoviocytes. Results WL attenuated RA symptoms in CIA mice as evidenced by delayed disease onset, decreased joint swelling, less bone invasion and lower cumulative incidence. WL also reduced the Th17/Treg cell ratio in the spleen and colon and improved the intestinal barrier by inhibiting colonic inflammation and up-regulating tight junction proteins ZO-1 and Occludin. Concomitantly, WL reversed the RA-induced dysbiosis of gut microbiota and markedly elevated SCFAs levels, thereby restoring intestinal barrier integrity and re-establishing Th17/Treg homeostasis. FMT significantly ameliorated arthritis, further verifying the role of the gut microbiota-joint axis in the anti-arthritic effect of WL. However, FMT only partially mimicked the efficacy of WL, which was comparable to that of WL in ABX mice, indicating an additional microbiota-independent pathway. Additionally, WL directly inhibited the proliferation and migration of both MH7A and RA-FLS cells and induced G0/G1 cell cycle arrest. Transcriptomic profiling revealed that WL up-regulated p53-pathway genes including MDM2 and CDKN1A. Subsequently, pharmacologic blockade and genetic knockdown of the p53 axis abolished WL-induced decreases in cell viability and EdU positive cells and reversed the up-regulation expression of CDKN1A and HMOX1. Mechanistically, WL directly bound to p53, disrupted the p53-MDM2 interaction, suppressed p53 ubiquitination and proteasomal degradation, thereby stabilizing and upregulating p53 expression. Conclusion WL mitigated CIA in mice by modulating gut microbiota-SCFAs-Th17/Treg axis and inhibiting proliferation of RA-FLS via p53 pathway. The findings provide the pre-clinical foundation for the development of WL as an anti-RA agent.

INTRODUCTION:Frankincense Essential Oil (FREO) has demonstrated curative potential in Ulcerative Colitis (UC) patients. However, the inherent instability of FREO results in its relatively low bioavailability. Therefore, the present study aimed to develop a novel oral O/W type FREO Submicron Emulsion Formulation (FREO-SE). This was achieved by encapsulating FREO within submicron emulsion droplets, with the further objective of elucidating the anti-UC efficacy of FREOSE. METHODS:A single-factor experimental approach was employed to screen the formulation, dosage, and preparation process of FREO-SE. Subsequently, the Box-Behnken Design (BBD) was utilized to optimize the submicron emulsion preparation procedure. The quality of the prepared emulsion was evaluated. Finally, a comparative analysis of the anti-ulcerative colitis efficacies of FREO and FREOSE was conducted using a UC mouse model. The mechanism of action of FREO-SE was further examined through immunohistochemistry, with the ultimate goal of enhancing the stability of FREO and elucidating its therapeutic effects on ulcerative colitis. RESULTS:The optimal formulation and manufacturing process for FREO-SE were established, and the particle size, PDI, and Zeta potential were characterized, with values of 105.09 ± 1.27 nm, 0.30 ± 0.02, and -37.43 ± 0.97 mV, respectively, confirming the successful preparation of FREO-SE. In DSS-induced UC mice, FREO-SE significantly reduced the DAI score compared with the DSS group. The weight loss of the FREO-SE-H group mice was significantly reduced (p < 0.001), and the shortening of colon length was significantly reduced (p < 0.001). Serum TNF-α and IL-6 levels were significantly reduced (p < 0.001), thereby alleviating colonic tissue lesions. The expression of p-ERK and p-P65 in colon tissue was significantly reduced (p < 0.001). In conclusion, FREO-SE inhibited the levels of p-ERK and p-P65 in MAPK and NF-κB signaling, and demonstrated a definite therapeutic effect in a mouse model of ulcerative colitis. DISCUSSION:This study confirmed that the FREO-SE formulation notably potentiates the therapeutic efficacy of FREO against UC, with its mechanism underlying modulation of the MAPK/NF-κB inflammatory signaling pathway. CONCLUSION:The preparation process of FREO-SE is characterized by stability, simplicity, and controllability, endowing it with excellent stability. FREO-SE exhibits a protective effect against DSSinduced UC in mice and demonstrates significant efficacy in the ulcerative colitis mouse model.

Abstract Ubiquitin-specific peptidase 22 (USP22), a deubiquitinase and component of the “Death-By-Cancer” gene signature, is overexpressed in many cancers and associated with recurrence, therapy resistance, and poor prognosis. Although USP22 has recently been implicated in tumor immune evasion, its role in KRAS-driven lung cancer and antitumor immunity has remained unclear. Here, we aimed to investigate the role of USP22 in tumor progression and antitumor immunity in KRAS-driven lung cancer. By immunohistochemistry analysis, USP22 was found to be highly expressed in nearly all human KRAS-mutant lung adenocarcinomas and in early-stage KRAS-induced mouse tumors, where its strong correlation with the proliferation marker Ki67 highlights USP22 as a potentially important mediator of KRAS-driven tumor biology and reinforces its promise as a therapeutic target. To define USP22’s role in tumor progression and immune regulation, we generated a conditional Usp22 knockout (Usp22-KO) model in KRASG12D; Tp53-/- (KP) mice. Usp22 loss significantly suppressed tumor growth, prolonged survival, and promoted tumor differentiation, accompanied by reduced KRAS pathway activity and partial restoration of p53 signaling. Spatial transcriptomics, RNA sequencing, and multiplex immunofluorescence revealed that Usp22 deletion reprogrammed the tumor microenvironment by decreasing CD206+ M2 macrophages, reducing TGF-β1 levels, limiting angiogenesis, increasing CD8+ T cell infiltration, and diminishing FOXP3+ regulatory T cells. Mechanistically, Usp22-KO altered gene expression and protein stability, reducing c-Myc, PD-L1, TGF-β1, and secreted protein acidic and rich in cysteine (SPARC); disruption of the TGF-β1-SPARC axis led to a marked depletion of immunosuppressive PD-L1^+ M2 macrophages. Functionally, Usp22 deletion substantially enhanced sensitivity to anti-PD-L1 immune checkpoint blockade in KP lung cancer. Collectively, these findings reveal USP22 as a critical driver of tumor progression and immunosuppression in KRAS-mutant, TP53-deficient lung cancer and demonstrate that targeting USP22 has significant therapeutic potential for overcoming resistance to immune checkpoint inhibitor therapy. Citation Format: Keqiang Zhang, Ching Ouyang, jinhui Wang, Wendong Li, Walter Tsark, Yuanyuan Gao, Mingxiao Yang, Aimin Li, Colt Egelston, Crystal Marconett, Dan Raz. Targeting USP22 reprograms the tumor microenvironment and sensitizes KRAS-mutant, TP53-Null Lung cancer to immunotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 2909.

Cisplatin, a chemotherapeutic drug, produces severe nephrotoxicity and at present, there are no effective drugs to clinically prevent or treat it. In the Dai nationality, BaiYangJie is used to treat poisoning caused by chemicals or drugs. Previous research has demonstrated that the methanolic extract of BaiYangJie (MEAG) can treat cisplatin-induced nephrotoxicity; however, its mechanism of action remains unclear. The aim of the present study was therefore to identify the potential mechanism of action of MEAG in cisplatin nephrotoxicity using a combination of network pharmacology and serum metabolomics. Initially, network pharmacology analysis was used to identify hub targets and signalling pathways involved in the renoprotective effects of MEAG. Subsequently, plasma metabolomics profiling utilising ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry technology revealed key metabolic alterations and pathway modulations associated with MEAG treatment. Finally, integrated analysis uncovered key molecular mechanisms, which were subsequently validated by western blotting and immunohistochemistry. A total of 13 endogenous metabolites were identified in serum metabolomics, primarily involved in phenylalanine metabolism and in the biosynthesis of phenylalanine, tyrosine and tryptophan. The treatment of MEAG in cis-induced acute kidney injury primarily involved regulating the inflammatory response, responses to lipid and chemical stress, the FoxO signalling pathway, arachidonic acid metabolism and the NF-κB signalling pathway. Animal experiments showed that MEAG can inhibit inflammation and the expression of the migration inhibitory factor (MIF)/NF-κB pathway. Through integrated network pharmacological analysis, metabolomic profiling and experimental validation, it was systematically elucidated that MEAG exerted its therapeutic effects through dual regulatory mechanisms: Suppressing inflammatory responses by inhibiting overactivation of the MIF/NF-κB signalling pathway and restoring phenylalanine metabolic homeostasis. These findings thus provide a mechanistic foundation for developing targeted therapeutic strategies against chemotherapy-associated nephrotoxicity.

Abstract Background Psoriasis is an immune-mediated chronic inflammatory skin disease. Existing therapies have limitations, necessitating the development of new treatment approaches. Compound Baixianpi Formula (FFBXP) is a clinically effective topical Chinese herbal formula, but its material basis and mechanism of action remain unclear. Materials and methods FFBXP components were identified using UHPLC-Q-Orbitrap HRMS. Core targets and pathways were screened by integrating network pharmacology and transcriptomics analyses. The effects of FFBXP on skin lesions, histopathology, oxidative stress, and inflammatory mediators (IL-17A, IL-23, TNF-α) were evaluated using a IMQ mouse psoriasis model and a TNF-α-stimulated HaCaT cell model. Key mechanisms were validated through molecular docking, qRT-PCR, Western blot, and immunofluorescence techniques. Results Forty-two active components were identified in FFBXP. In vivo experiments demonstrated that FFBXP significantly improved erythema and skin lesion infiltration in psoriatic mice, while reducing levels of inflammatory cytokines (IL-17A, IL-23, TNF-α) and oxidative stress markers (MDA). In vitro experiments confirmed that FFBXP dose-dependently inhibited TNF-α-induced proliferation in HaCaT cells, reduced inflammatory cytokine levels, mitigated oxidative stress, and promoted apoptosis. Network pharmacology and transcriptomics analysis indicated its mechanism involves IL-17, PI3K-AKT, and JAK2-STAT3 signaling pathways. Molecular docking showed that six core active ingredients (berberine, resveratrol, quercetin, catechin, kaempferol, and osthol) had good binding activity with key target proteins. Further mechanism validation revealed FFBXP significantly decrease IL-17A expression and inhibited phosphorylation of downstream PI3K, AKT, JAK2, and STAT3 proteins. Conclusion This study employs a combined strategy of network pharmacology, transcriptomics, and experimental validation to elucidate for the first time that FFBXP exerts its anti-psoriasis effects by targeting IL-17A and collaboratively inhibiting two key signaling pathways: PI3K-AKT and JAK2-STAT3.

BACKGROUND:Amomum kravanh Pierre ex Gagnep. (BDK) is a Zingiberaceae plant traditionally widely used as a sweet fragrance, and commonly also utilized in minority medicine for various kidney diseases, especially chronic kidney disease (CKD) in Tibetan and Mongolian medicine. However, the underlying mechanisms by which it confers renal protection remain to be fully clarified. PURPOSE:To investigate the renal protective mechanism of which BDK's essential oil exerts in rats with CKD induced by adenine and 5/6 nephrectomy. METHODS:Rat models of adenine and 5/6 nephrectomy chronic nephropathy were established, and the therapeutic effects were evaluated by detecting the blood biochemical levels and H&E-/Masson staining and fiber-related factors. Then, the chemical composition of BDK's essential oil and blood components were analyzed using GC-MS. The efficacy of eucalyptol was evaluated by adenine and 5/6 nephrectomy CKD model, with mechanistic studies conducted using RNA-seq, western blot, and metabolomic approaches. RESULTS:The blood biochemical levels and histopathological analyses (H&E-/Masson's staining) revealed that the BDK's essential oil significantly enhanced renal function and ameliorated kidney tissue fibrosis. Furthermore, GC-MS analysis identified 33 components in the essential oil of BDK, with eucalyptol being the predominant chemical component at 74.07 %. Eucalyptol is capable of entering the bloodstream in its prototypical form. Then, the efficacy and mechanism of eucalyptol were confirmed by adenine/5/6 nephrectomy CKD models, and based on RNA-seq analysis, we found that eucalyptol could significantly improve kidney function and fibrosis of kidney tissues by blocking TGF-β/smad and NF-κB pathways and inhibit ferroptosis through the Nrf2/HO-1 signaling pathway. CONCLUSION:Both BDK's essential oil and its main constituent, eucalyptol, exhibited protective effects against CKD. They both ameliorated oxidative stress, inflammation, and fibrosis in adenine/5/6 nephrectomy rats. Eucalyptol is implicated in ferroptosis and regulation of renal fibrosis via the Nrf2/HO-1 pathway.

Liver cancer, a malignancy with high global incidence and mortality, currently relies on surgical resection, radiotherapy, and chemotherapy, all of which face significant limitations, necessitating novel therapeutic strategies. Mentha (ME), a medicinal and edible herb, has demonstrated antioxidant, anti-inflammatory, and broad-spectrum anticancer activities, yet its molecular mechanisms against liver cancer remain unclear. This study will comprehensively explore the anti-liver cancer mechanisms of ME and its key bioactive constituent, diosmetin (Dio). A multi-disciplinary approach, which incorporates network pharmacology, molecular docking, and molecular dynamics simulations, was adopted in this study to thoroughly explore the bioactive components of ME and the mechanisms through which they exert anti-liver cancer effects. Functional validation was conducted through CCK-8 viability assays, clonogenic survival assays, scratch wound healing, Transwell migration assays, Western blotting, immunofluorescence, and TUNEL apoptosis assays in human liver cancer cell lines (HepG2 and HuH-7). ME exhibited potent anti-Liver cancer activity, significantly suppressing cell viability, proliferation (CCK-8/clonogenic assays), and migration (scratch/Transwell assays, P < 0.01), while downregulating metastasis-related proteins MMP2/MMP9 (Western blot/immunofluorescence, P < 0.01). Network pharmacology identified TP53 (p53), TNF, CASP3 (caspase3), IL6, and IL1B as core targets. Based on the results of molecular docking (ΔG < - 4 kcal/mol) and molecular dynamics simulations (maximum ΔTotal), Dio was prioritized for subsequent experimental validation. Further validation demonstrated Dio's multi-modal efficacy: GO/KEGG analysis revealed its dual action via p38/MAPK signaling and apoptosis pathways, corroborated by upregulated pro-apoptotic markers (p53, caspase3, Bax, p38) and downregulated Bcl2 (P < 0.01), alongside TUNEL-confirmed apoptosis induction (P < 0.01). This study is the first to demonstrate that ME and its active compound Dio inhibit liver cancer progression via multi-target regulation of the p38/MAPK pathway, providing a theoretical foundation for developing ME-based natural therapeutics against liver cancer.

ETHNOPHARMACOLOGICAL RELEVANCE:Non-alcoholic fatty liver disease (NAFLD) is one of the most prevalent liver diseases worldwide, with an estimated global prevalence of 30 %. An imbalance in lipid metabolism leads to the accumulation of lipotoxic lipids, inducing cellular stress, activating the NLRP3 inflammasome, and triggering apoptotic cell death. This cascade stimulates inflammation, driving NAFLD progression. Morus nigra L. is the only black mulberry species native to China. In traditional Uygur medicine, its fruit is valued for its hepatoprotective and lipid-lowering effects. AIM OF THE STUDY:The mechanism by which mulberry extract (ME) alleviates NAFLD remains unclear. This study aimed to investigate the hepatoprotective and anti-inflammatory effects of ME. MATERIALS AND METHODS:Network pharmacology was employed to predict the active components of ME and their potential target genes. Liver function markers (ALT, AST) and lipid profiles (TG, TC) were assessed using commercial assay kits. The therapeutic mechanism of ME against NAFLD was elucidated through an integrated approach combining transcriptomic and metabolomic analyses in a mouse NAFLD model. RESULTS:UPLC-QTOF-MS analysis identified 131 active ingredients in ME. Network pharmacological analysis identified Akt1, Pparg, and Pparα as core targets. High-dose ME treatment markedly improved liver function, reducing ALT levels by 50 % and AST levels by 44 %, while also lowering hepatic TG by 22 % and TC by 15 %. Transcriptome analysis revealed that ME ameliorated NAFLD through AMPK/PPAR-γ/NF-κB axis. Metabolomic analysis demonstrated the involvement of unsaturated fatty acid and steroid hormone biosynthesis in NAFLD metabolism. CONCLUSIONS:Our study demonstrates that ME alleviates NAFLD progression by regulating the AMPK/PPAR-γ/NF-κB signaling axis. However, these findings are based on preclinical animal studies, and further investigation is required to determine the clinical applicability of these effects in human patients.

Although lavandula angustifolia essential oil (LEO) has demonstrated anti-inflammatory properties, its mechanism in alleviating chronic prostatitis (CP) remains unclear. This study aimed to explore the pharmacological basis and molecular mechanisms of LEO's effect on CP. The chemical composition of LEO was analyzed using Gas Chromatography-Mass Spectrometry (GC-MS), identifying 15 active ingredients. A carrageenan-induced rat CP model was used to evaluate the impact of LEO on prostate index, inflammatory markers, and histopathological changes. The results showed that LEO significantly reduced the prostate index, inhibited inflammatory markers, and improved tissue damage. Multi-omics analysis, including transcriptome sequencing, Weighted Gene Co-expression Network Analysis (WGCNA), network pharmacology, and metabolomics, revealed that LEO modulates key pathways such as phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) and nuclear factor kappa-B (NF-κB). Molecular distillation experiments and in vitro screening identified linalool and linalyl acetate as the main anti-inflammatory components of LEO. Molecular docking (MD) and molecular dynamics simulations (MDS) confirmed strong binding affinities of these compounds to AKT1 and Nuclear Factor Kappa B subunit P65 (P65), which were further validated by Surface Plasmon Resonance (SPR). Immunohistochemistry (IHC) and Western blot (WB) analyses confirmed the inhibition of cyclooxygenase-2 (COX-2), phosphatidylinositol 3-kinases (p-PI3K), phosphorylated protein kinase-B (p-AKT), and phosphorylated Nuclear Factor Kappa B subunit P65 (p-P65) expression. In conclusion, this study demonstrates that LEO alleviates CP through the PI3K/AKT/NF-κB pathway, primarily mediated by linalool and linalyl acetate, suggesting its potential as a therapeutic agent for CP.