Metabolic-associated fatty liver disease (MAFLD) progresses via a vicious cycle of "lipid dysregulation-ceramide-inflammation-oxidative stress-ferroptosis," with sodium palmitate (PA) as a key mediator of hepatic lipotoxicity. To screen ameliorative natural compounds, we performed high-throughput screening of 236 traditional Chinese medicine-derived compounds using PA-induced AML-12 hepatocytes, identifying biochanin A (BCA)-a major isoflavone in chickpeas-as a potent protector against hepatocyte death. We validated BCA's effects in vitro (PA-induced AML-12 cells) and in vivo (high-fat diet-induced MAFLD mice, 25/50 mg/kg BCA), combined with IRE1α agonist IXA4 rescue experiments and multi-omics analyses. A novel finding is that BCA directly binds IRE1α (via LEU23/CYS91, validated by molecular docking and 100-ns MD simulations) and specifically inhibits the IRE1α-SPT-ceramide axis. This downregulates SPTLC1/SPTLC2 (ceramide synthesis rate-limiting enzymes), normalizes hepatic C16/C24 ceramide levels, suppresses IL-1β/IL-6 production, restores mitochondrial OXPHOS function, reduces ROS/lipid peroxidation, and inhibits ferroptosis. Concurrently, BCA treatment was associated with alterations in gut microbiota composition (enriching Bacilli, reducing pro-inflammatory Coriobacteriia), enhanced intestinal barrier function, and reduced LPS translocation, which may contribute to mitigating hepatic inflammation. Notably, IXA4 completely reversed BCA's protective effects. In conclusion, BCA ameliorates MAFLD by inhibiting the IRE1α-SPT-ceramide axis to block the pathological cascade. In parallel, BCA treatment is associated with alterations in gut microbiota composition, improved intestinal barrier integrity, and reduced systemic inflammation, suggesting that the gut-liver axis may be involved in its protective effects.

Adzuki bean saponin (ABS) has shown therapeutic potential in improving obesity, but its underlying mechanism and active saponin components are still unknown. This research estimated the anti-obesity effect of ABS in obesity using a mouse model. Advanced research methods such as spatial metabolomics, transcriptomics, proteomics, serum pharmacology, and network pharmacology were employed to study the molecular mechanisms of ABS and its bioacitve saponins. ABS achieved anti-obesity effects by improving HFD-induced body weight gain, abnormal lipid profiles and liver dysfunction liver lipid accumulation, and increased adipose tissue. Liver spatial metabolomics suggested a potential involvement of ABS in modulating lipid metabolism via alterations in the arachidonic acid pathway. The results of transcriptomics, proteomics, and immunohistochemistry indicated that ABS could achieve anti-obesity effects through activating the Wnt/β-catenin signaling pathway. Based on our previous study and serum pharmacology analysis revealed that soyasaponin Ba is a key saponin in ABS that achieves anti-obesity effects. Similarly, soyasaponin Ba can achieve anti-obesity effects by activating Wnt/β-catenin signaling. ABS and its active saponin Soyasaponin Ba can achieve anti-obesity effect by up-regulation of the Wnt/β-catenin signaling pathway. This understanding of this mechanism provides valuable insights for developing drugs or health supplements based on adzuki beans to combat obesity.

BACKGROUND:Malignant melanoma remains a major challenge in oncology, largely due to the limited efficacy and significant toxicity associated with treatments such as temozolomide (TMZ). Si-Jun-Zi-Tang (SJZT), a traditional Chinese medicine formula with documented anti-cancer properties, has shown potential in enhancing chemotherapy effectiveness and reducing treatment-related toxicity. PURPOSE:This study investigated the role of SJZT in improving TMZ treatment outcomes in melanoma. METHODS:The effects of SJZT on TMZ treatment outcomes were evaluated in melanoma cell lines and mouse models. Cell proliferation, apoptosis, DNA damage, and cell cycle distribution were assessed using CCK-8 assays, Annexin V/PI staining, γ-H2AX immunofluorescence, and flow cytometry, respectively. In vivo antitumor efficacy and hepatotoxicity were evaluated in melanoma-bearing mice. Mechanistic insights were explored via network pharmacology and RNA sequencing, and validated by RT-qPCR, Western blotting, and rescue experiments. RESULTS:SJZT enhanced TMZ-induced inhibition of cell proliferation, apoptosis, and DNA strand breaks in melanoma cells. SJZT also attenuated TMZ-induced G2/M arrest, promoting mitotic catastrophe. In mouse models, SJZT enhanced the anti-tumor effects of TMZ and mitigated TMZ-induced hepatotoxicity. Mechanistically, network pharmacology and experimental validation revealed that simultaneous inhibition of Chk1 and promotion of MGMT proteasomal degradation mediated the enhanced anti-melanoma effects of TMZ with SJZT. RNA sequencing and experimental validation demonstrated that TMZ upregulated the bile salt transporter Slc10a1/Ntcp and downregulated detoxifying enzymes Ugt1a1 and Slco1b2/Oatp2, disrupting bile acid homeostasis. Notably, SJZT co-administration reversed these changes, restoring bile acid balance. CONCLUSION:These findings reveal that SJZT can enhance TMZ's therapeutic efficacy while reducing hepatotoxicity, supporting its use as a promising adjuvant in melanoma treatment.

Current disease-modifying therapies for multiple sclerosis (MS) primarily target peripheral immune responses but exhibit limited efficacy in mitigating the compartmentalized neuroinflammation driven by central nervous system (CNS)-resident microglia. By integrating clinical sample analysis with experimental autoimmune encephalomyelitis (EAE) model studies, we have demonstrated that the proviral integration site for Moloney murine leukemia virus 1 (PIM1) is significantly upregulated, particularly in microglia, in both MS patients and the spinal cords of EAE mice. This upregulation positively correlates with disease severity and levels of proinflammatory cytokines such as IL-1β, TNF-α, and IL-6. Utilizing a multimodal research approach-including pharmacological inhibition (SMI-4a), genetic knockdown, RNA sequencing, and HIS-SIM super-resolution imaging-we confirmed that PIM1 inhibition effectively attenuates neuroinflammatory responses, improves clinical symptoms in EAE mice, and promotes activation of the mitophagy pathway while suppressing inflammation-related molecules. Mechanistically, PIM1 enhances the phosphorylation of dynamin-related protein 1 (Drp1) at Ser616 while suppressing its phosphorylation at Ser637, which disrupts LC3-mitochondria colocalization and autophagosome-lysosome fusion. This leads to mitophagy dysfunction, loss of mitochondrial membrane potential, and accumulation of reactive oxygen species. Notably, the combined administration of PIM1 and Drp1 inhibitors did not yield synergistic therapeutic effects, suggesting that PIM1 likely functions as an upstream master regulator of Drp1. These findings not only elucidate the molecular mechanism by which PIM1 interacts with Drp1 to regulate microglial activation and mitophagy but also establish PIM1 as a promising CNS-intrinsic therapeutic target for restoring mitochondrial homeostasis in MS.

Non-alcoholic fatty liver disease (NAFLD) accounts for remarkable burden of death and costs worldwide with no recommended pharmacological intervention for the clinical management. This study aimed to investigate the efficacy and underlying mechanisms of rhubarb-derived polysaccharides (RP) in mitigating high-fat diet (HFD)-induced NAFLD and to analyze the primary monosaccharide components of RP. Forty male C57BL/6 mice were subjected to a dietary intervention consisting of either a high fat or chow diet for a duration of 12 weeks. RP (270, 540 mg·kg-1·d-1) was administered to the mice for 4 consecutive weeks from the 9th week. Various assessments were conducted, including histopathological examination, liver transcriptome analysis, non-targeted metabolomics analysis, and evaluation of protein expressions related to lipid and bile acid metabolism. This study found RP demonstrate a protective effect on the livers of NAFLD mice by inhibiting lipid accumulation and reducing hepatocyte inflammatory damage. The metabolomics analysis of multi-tissues revealed that the RP exert a hepatoprotective effect against NAFLD by restoring the altered bile acids (BAs) and fatty acids (FFAs) metabolism through the improvement of BA transporter, nucleus hormone receptor, lipogenesis protein, FFA transporter, and lipolysis proteins. Hence, RP may serve as a potential therapeutic agent for NAFLD.

This study aimed to explore the efficacy and mechanisms of raspberry (Rubus idaeus L. fruit) aqueous extract (RE) in alleviating high-fat diet (HFD)-induced metabolic-associated fatty liver disease (MAFLD). The MAFLD mouse model was established to examine the effects of RE through liver transcriptome and metabolomics analysis. In this study, RE supplementation significantly alleviated HFD-induced liver injury, hepatosteatosis, inflammation, and insulin resistance. Liver transcriptome analysis demonstrated that RE supplementation favorably regulated signaling pathways involved in fatty acid metabolism and inflammation, including the AMPK signaling pathway, PPAR signaling pathway, apoptosis, etc. Furthermore, the injection of compound C, an antagonist of AMPK, notably reversed the hepatoprotective effects of RE, evidenced by increased lipid profile levels, accelerated fatty acid-related gene disorder, and increased positive tunnel staining area. Furthermore, liver metabolomics analysis demonstrated that RE treatment led to substantial enrichment of the liver tissue metabolite umbelliferone (UMB), which has the potential to ameliorate lipid accumulation and hepatocyte injury through the AMPK signaling pathway. In summary, RE intervention mitigated HFD-induced liver dysfunction in mice, with UMB likely being the primary component responsible for its therapeutic efficacy in the liver. In addition, this study provided new insights, suggesting that RE could be used as a promising therapeutic approach for modulating MAFLD via apoptosis and the AMPK/PPARα signaling pathway.

Renal cell carcinoma (RCC) is the most common kidney cancer. Despite advances in treatment, current therapeutic strategies are often limited by side effects, drug resistance, and low response rates, necessitating alternatives for RCC treatment. Deoxyelephantopin (DEO), a sesquiterpene lactone from Elephantopi Herba, has demonstrated anticancer properties in multiple cancer models; however, its effects on RCC remain unknown. This study aimed to investigate the anti-RCC effects of DEO and its underlying molecular mechanisms. Human RCC cell lines (786-O, Caki-1, A498) and a murine RCC cell line (RENCA) were used for in vitro assays. Results revealed that DEO dose-dependently inhibited cell viability and colony formation in 786-O, Caki-1, A498, and RENCA cells, while also inducing apoptosis in 786-O and Caki-1 cells. A RENCA allograft mouse model was used for in vivo assays. DEO significantly suppressed tumor growth without causing notable changes in body weight, organ coefficients, or serum biochemical markers (ALT, AST, BUN, Cr). Network pharmacology analysis predicted the PI3K/AKT signaling pathway as a key mediator of DEO's anti-RCC effects. Western blotting showed that DEO downregulated the expression of EGFR, p-EGFR (Tyr1068), PI3K p110α, p-Akt (Ser473), mTOR, p-mTOR (Ser2448), p-p70S6K (Thr389), 4E-BP1, p-4E-BP1 (Thr37/46), HIF-1α, and Bcl-2. Overactivation of AKT attenuated DEO's inhibitory effects on cell viability in 786-O cells. In conclusion, this study is the first to demonstrate that DEO exerts anti-RCC effects in both cellular and animal models, primarily through inhibition of the PI3K/AKT pathway. These findings suggest that DEO holds promise as a lead compound for RCC management.

BACKGROUND:Alisol B (AB) has been demonstrated to be a potential lead compound in improving obesity-related metabolic disorders. Nevertheless, the effects and mechanisms of AB on Metabolic dysfunction-associated steatotic liver disease (MASLD) remain unclear. PURPOSE:This study aimed to investigate the improvement of AB on MASLD and explore the intricate mechanism involving gut microbiota and liver metabolism. MATERIALS AND METHODS:The MASLD mice model was established by feeding a high-fat diet and oral treatment with AB. The effects of AB on lipid metabolism in MASLD were initially measured. Subsequently, 16S rRNA gene sequencing, untargeted metabolomics combined with network pharmacology analysis was used to unveil the potential mechanism of AB on MASLD. A series of molecular biology experiments was conducted to confirm the results of the multi-omics analysis and to elucidate the key mechanism. RESULTS:AB attenuated liver steatosis and improved liver injury in MASLD mice. AB treatment improved the diversity of gut microbiota and increased the abundance of Akkermansia, Escherichia-Shigella, and Muribaculu in MASLD mice. Based on correlation analysis between differential intestinal microbiota and metabolites, metabolites involving sodium oleate, helleolate acetate 3-acetate and oxaminate were identified as key metabolites. In addition, integrating metabolomics and network pharmacology showed that AB alleviated MASLD by regulating the purine metabolism pathway and de novo fatty acid biosynthesis. Then, we focused on the role of purine metabolism in the treatment of MASLD by AB. Notably, AB inhibited the urine acid level in serum and liver of MASLD mice and hepatic XO activity and expression. AB markedly reduced the hypoxanthine and allantoin levels, increased the inosine level in the livers of MASLD mice, indicating that AB significantly reversed the dysfunction of hepatic purine metabolism in MASLD. Moreover, Molecular docking and surface plasmon resonance (SPR) results demonstrated that AB directly binds to XO. Overexpressing XO abolished the effect of AB in lipid accumulations in AML-12 cells. AB may alleviate MASLD by directly targeting XO to inhibit purine metabolism disorders in the liver. CONCLUSION:Our results demonstrate that AB treatment attenuates HFD-induced MASLD through dual mechanisms involving gut microbiota modulation and restoration of hepatic metabolic homeostasis. Comprehensive mechanistic analysis revealed that AB ameliorates hepatic steatosis and corrects purine metabolism dysregulation in MASLD pathogenesis through specific inhibition of XO. These findings provide novel mechanistic insights into the hepatoprotective properties of AB and establish its therapeutic potential for MASLD intervention.

Rapid urbanization, medication, and modern dietary patterns are the main challenges leading to impaired fertility, with a lack of effective therapies. Emerging evidence suggests that reproductive disorders may be closely associated with intestinal damage or occasionally worsen by the side effects of medications. Therefore, the development of dietary supplements as alternatives is crucial for intestinal-linked reproductive health. Milk and dairy products are essential in dietary nutrition with great potential functional ingredients. Bifidobacterium animalis subsp. lactis NJ241 (NJ241), a promising probiotic isolated from naturally fermented bovine milk, remains underexplored. This study aims to investigate the molecular mechanisms of NJ241 on colitis and its associated reproductive disorders. The biomarker microbes and their correlated metabolites were further explored by 16S rRNA sequencing and metabolomics. Hematological analysis and histopathological examination were applied for conjoined identification. Results indicated that NJ241 effectively restored the expression levels of Claudin-2 and MLCK1, reducing intestinal permeability. Multi-omics results further revealed that NJ241 may effectively improve gut barrier integrity by increasing the abundance of Akkermansia muciniphila and its metabolite trans-ferulic acid. This effect was accompanied by a reduction in the pro-inflammatory cytokine IL-6 in both serum and testicular tissue, mediated through the TLR4 signaling pathway. Consequently, the restoration of microbiota homeostasis and a systemic reduction in inflammation rescued testicular spermatogenesis, which was impaired by colitis. The current findings consistently elucidated the potential molecular mechanism by which NJ241 ameliorates colitis-linked reproductive disorders through the gut-testis axis. Additionally, NJ241 demonstrates promise as a probiotic supplement for the development of fortified dairy products and provides strong evidence for the potential reproductive health benefits of naturally fermented bovine milk.

Curcumin is renowned for anti-inflammatory, antioxidant and hepatoprotective effects, and has been implicated in the amelioration of obesity and diabetes. Notwithstanding its considerable therapeutic potential, the clinical utility of curcumin is hampered by its suboptimal bioavailability, due to poor aqueous solubility and chemical instability. Consequently, the development of strategies to enhance the aqueous solubility, stability, and ultimately, the bioavailability of curcumin has been a focal point of intense research. This study harnessed tetrahedral framework nucleic acids (tFNAs), a relatively simple DNA nanostructure, to encapsulate curcumin. Meanwhile, novel aptamers for liver-specific targeting were acquired by SELEX (Systematic Evolution of Ligands by Exponential Enrichment) method. By capitalizing on the unique properties of aptamers and tFNAs, an aptamer-mediated liver-targeted curcumin delivery system was constructed, with the goal of providing a more efficacious therapeutic approach for non-alcoholic fatty liver disease (NAFLD). This innovative delivery platform has not only markedly improved the solubility and stability of curcumin but has also significantly bolstered its therapeutic efficacy in the context of NAFLD. This research not only offers a novel approach for the delivery of curcumin but also presents a new therapeutic modality for NAFLD. Moreover, the implications of this research extend beyond curcumin, offering a blueprint for the liver-targeted delivery of other drug molecules.

Metabolic dysfunction-associated steatotic liver disease (MASLD) is a prevalent global health issue characterized by excessive fat accumulation in the liver, often linked to obesity and metabolic syndrome. Despite advancements in understanding its pathogenesis, effective therapeutic strategies remain limited. This study investigates the potential of Alisol B, a natural compound from traditional Chinese medicine, in modulating lipid metabolism and autophagy in hepatocytes. We employed a combination of in vivo and in vitro approaches, including mouse models, cell culture assays, and transcriptomic profiling, to evaluate Alisol B's therapeutic efficacy against MASLD and elucidate its underlying mechanisms. Our findings reveal that Alisol B significantly reduces lipid accumulation and enhances fatty acid metabolism by upregulating Ces2a, a key regulator of lipid catabolism, as confirmed by RNA sequencing and Western blot analyses. Additionally, transcriptomic analysis indicates that Alisol B activates critical signaling pathways related to fatty acid metabolism and autophagy, including AMPK signaling. Importantly, in vitro studies demonstrate that Alisol B effectively reduces triglyceride levels in hepatocytes without compromising cell viability. Pharmacological inhibition of Ces2a further underscores its essential role in mediating Alisol B's therapeutic effects. These results suggest that Alisol B holds promise as a novel therapeutic agent for MASLD, warranting further exploration of its clinical applications and potential as a targeted treatment for metabolic disorders.

RNAi therapeutics can potentially address undruggable diseases. However, their full potential is stymied by delivery challenges. An siRNA delivery platform by remodeling the human U4 small nuclear ribonucleoprotein complex (snRNP) is developed. This remodeled protein-siRNA complex (SmiRNP) serves as a biocompatible, modular, and customizable platform for attaching desired functional modules to overcome delivery hurdles. Here, it is demonstrated that the SmiRNP complex, by using siRNA against KRAS as a proof of principle, can deliver the siRNA into cells and protect it from nuclease degradation, allowing the siRNA to knockdown KRAS mRNA and protein levels, reduce cancer cell viability, and suppress tumor growth in vivo. This general strategy allows many more modular combinations and would enable the delivery of a wide spectrum of RNAi therapeutics.

BACKGROUND:Oncogenic mutations in the BRAF gene, particularly the V600E mutation, are present in roughly half of metastatic cutaneous melanoma cases. BRAF inhibitors (BRAFi) have shown significant clinical efficacy; however, their long-term effectiveness is frequently limited by the development of resistance. Parthenolide, a sesquiterpene lactone derived from medicinal herbs, has previously been reported to exhibit anti-melanoma effects and to disrupt signaling pathways associated with BRAFi resistance. PURPOSE:This study aimed to evaluate the potential of parthenolide to overcome resistance to BRAFi in melanoma. STUDY DESIGN AND METHODS:The effects of parthenolide on BRAFi-resistant melanoma cells were assessed using cell viability, apoptosis, and cell cycle assays. In vivo efficacy and safety were evaluated in a BRAFi-resistant melanoma xenograft mouse model. Target identification and validation were performed using network pharmacology, molecular docking, surface plasmon resonance, and Western blotting. RESULTS:Parthenolide inhibited cell viability and induced apoptosis and S-phase cell cycle arrest in BRAFi-resistant melanoma cells. In vivo, parthenolide significantly suppressed tumor growth and reduced tumor weight in BRAFi-resistant melanoma xenografts without apparent toxicity. Mechanistically, parthenolide stably interacted with the N-terminal ATPase domain of Hsp90α and dose-dependently inhibited its ATPase activity. Hsp90α functions as a chaperone for several oncoproteins involved in signaling pathways driving BRAFi resistance. Parthenolide reduced the levels of several Hsp90α client proteins and inhibited their downstream pathways, including PI3K/Akt, Ras/Raf/MEK, and Src/STAT3, in BRAFi-resistant melanoma cells. Moreover, the Hsp90 activator tamoxifen attenuated the effects of parthenolide in these cells. CONCLUSION:Our findings provide the first evidence that parthenolide can overcome BRAFi resistance in melanoma, highlighting its potential as a lead compound for the development of Hsp90α inhibitors to manage BRAFi-resistant cutaneous melanoma.

Arsenic, a known environmental carcinogen, disrupts intestinal homeostasis, posing a significant threat to human health. Mitigating its toxic effects is crucial, and this study explores the potential of swim bladder sulfated glycosaminoglycan (SBSG) in achieving this. Our previous in vitro studies have shown that SBSG to ameliorate arsenic-induced damage in intestinal epithelial cells, but its in vivo effects remain elusive. The current investigation demonstrates that SBSG exhibits a beneficial prebiotic action in vivo, regulating gut microbiota, metabolites, and intestinal barrier function to counter arsenic's adverse effects. Specifically, SBSG regulates microbiota composition, suppressing pathogenic species like Alistipes and Candidatus_Saccharimonas while promoting beneficial ones such as Ruminococcus and Akkermansia. In the colon, SBSG fermentation enhances the production of short-chain fatty acids (SCFAs), leading to the upregulation of GPR43, GPR109A, and Olfr78 receptors. Additionally, SBSG strengthens the intestinal barrier by increasing the expression of Claudin-1, Occludin, and ZO-1, and enhances mucin gene expression (MUC-1 and MUC-2) to address chemical barrier disruptions. Immunologically, SBSG modulates the RORγt/Foxp3 pathway and the TLR4/MyD88/NF-κB signaling cascade, regulating the immune barrier. These findings suggest that SBSG could be a promising prebiotic candidate for maintaining intestinal health and may serve as a dietary supplement or adjunct in heavy metal detoxification therapies.

BACKGROUND:The tri-herb formulation GGC, consisting of Ganoderma (GL, the dried fruiting body of Ganoderma lucidum), Gastrodiae Rhizoma (GR, the dried tuber of Gastrodia elata), and Chrysanthemi Flos (CF, the dried flower head of Chrysanthemum morifolium), has shown clinical potential for managing sleep disorders. However, its pharmacological basis remains underexplored. PURPOSE:This study aimed to investigate the sedative-hypnotic effects and underlying mechanisms of an ethanolic GGC extract (GGCE). METHODS:Mouse models were used to evaluate the sedative-hypnotic effects of GGCE. Mechanistic investigations were performed using RNA sequencing, RT-qPCR, ELISA, and immunoblotting. RESULTS:GGCE administration produced dose-dependent sedative-hypnotic effects, including reduced locomotor activity, shortened sleep latency, and prolonged sleep duration. RT-qPCR analysis revealed that GGCE upregulated Gabrd mRNA, which encodes the delta subunit of the GABAA receptor, in brain tissue. Immunoblotting showed increased protein levels of Gabrd, phospho-PKA C (Thr197), and phospho-CREB (Ser133) following GGCE treatment. Additionally, ELISA results indicated that GGCE elevated levels of GABA (gamma-aminobutyric acid), cAMP (cyclic adenosine monophosphate), and BDNF (brain-derived neurotrophic factor) in brain tissue. CONCLUSIONS:This study demonstrates for the first time that GGCE exerts sedative-hypnotic effects in mice. These effects are associated with activation of the cAMP/PKA/CREB/BDNF signaling pathway and enhanced GABAergic signaling. These findings provide the first pharmacological justification for the use of GGC in treating insomnia and suggest GGCE's potential as a modern therapeutic agent for improving sleep.

Atopic dermatitis (AD) is a prevalent inflammatory skin disorder with limited treatment options, highlighting the need for alternative therapeutic strategies. Traditionally, black soybean oil, prepared from Glycine max through high-temperature dry distillation, has been used in Asian medicine to relieve AD symptoms. However, this preparation method poses safety concerns due to the potential formation of carcinogenic by-products. To address these limitations, we prepared black soybean extract (BSE) using supercritical CO2 extraction and investigated its pharmacological effects and underlying mechanisms in AD-related cell models. Gas chromatography-mass spectrometry analysis identified five major fatty acids in BSE. In LPS-stimulated RAW264.7 macrophages, BSE reduced the production of pro-inflammatory mediators. In TNF-α/IFN-γ-stimulated HaCaT keratinocytes, BSE downregulated the mRNA levels of AD-associated alarmins, decreased apoptosis, and increased junction protein expression, suggesting its skin barrier-protective effects. Integrated network pharmacology and RNA sequencing analyses predicted that TNF signaling and its downstream MAPK and NF-κB pathways play key roles in BSE's effects. RT-qPCR validated that BSE downregulated the mRNA levels of genes involved in pro-inflammatory responses, immune activation, and skin barrier impairment. Western blotting results demonstrated that BSE inactivated MAPK and NF-κB signaling molecules, including ERK, p38, JNK, p65, and IκBα. Collectively, our findings demonstrate, for the first time, that BSE has anti-inflammatory and skin barrier-protective potential, with the inhibition of TNF-MAPK/NF-κB signaling pathways involved in BSE's effects. This study suggests that BSE has the potential to be used for managing AD-related inflammation and skin barrier dysfunction.

BACKGROUND:Atopic dermatitis (AD) is characterized by both IgE- and non-IgE-mediated immune responses, as well as skin barrier dysfunction. Ginsenoside Rg1, tetrandrine, and icariin each exhibit distinct properties that may contribute to the management of AD. Ginsenoside Rg1 has demonstrated efficacy in mitigating IgE-mediated allergic rhinitis, while tetrandrine is known to suppress abnormal T-cell activation. Icariin has been shown to improve intestinal barrier function, which is crucial in conditions like AD. However, the potential effectiveness of the combined formula of these compounds, referred to as GTI, in treating AD remains unexplored. PURPOSE:This study aimed to investigate the anti-AD effects and mechanisms of GTI in a mouse model. METHODS:A calcipotriol (MC903)-induced AD-like dermatitis mouse model was used to evaluate the anti-AD effects of GTI. Dermatitis scores and mouse ear thickness were recorded to assess disease severity. Ear tissues, ear-draining lymph nodes, spleens and sera were collected for use in the investigation of the effects and mechanisms of action of GTI. RESULTS:Topical application of GTI significantly alleviated AD-like dermatitis in mice, as evidenced by decreased dermatitis scores, reduced ear thickening, and diminished epidermal and dermal thickness, along with lower levels of the inflammatory cytokines IL-1β and IL-4 in ear tissues. Unlike the positive dexamethasone, GTI had no significant toxicity in the model mice. Topical GTI lowered serum IgE levels and diminished the accumulation of eosinophils and mast cells in ear tissues of model mice, suggesting that GTI mitigates IgE-mediated allergic reactions. GTI significantly decreased the numbers of CD4+ T cells in ear tissues, ear-draining lymph nodes and the spleen, demonstrating its suppressive effect on hyperactive immune responses. The protein levels of ZO-1 and claudin-1, two tight junction proteins, were elevated in the ear tissues of mice treated with GTI, indicating a beneficial effect of this formula on skin barrier function. Additionally, GTI inhibited the activation of mitogen-activated protein kinases (MAPKs), as indicated by the downregulation of phospho-p38 (Thr180/182), phospho-ERK (Thr202/Tyr204), and phospho-JNK (Thr183/185) protein levels in mouse ear tissues. CONCLUSION:This study, for the first time, demonstrated that the topical application of GTI alleviates symptoms of AD without overt toxicity in a calcipotriol-induced AD mouse model. The anti-AD effects of GTI are associated with the suppression of allergic reactions, reduction of hyperactive immune responses, improvement of skin barrier function, and inhibition of MAPK activation. These findings suggest that GTI has the potential to be developed into a safe and effective treatment for AD.

Purpose: Atractylodes macrocephala Koidz is a widely used classical traditional Chinese herbal medicine, that has shown remarkable efficacy in cancers. Colorectal cancer (CRC) is the most common malignant tumor globally. Interferon (IFN)- gamma , a prominent cytokine involved in anti-tumor immunity that has cytostatic, pro-apoptotic, and immune-stimulatory properties for the detection and removal of transformed cells. Atractylenolides-II (AT-II) belongs to the lactone compound that is derived from Atractylodes macrocephala Koidz with anti-cancer activity. However, whether AT-II combined with IFN- gamma modulates CRC progression and the underlying mechanisms remain unclear. The present study aimed to elucidate the efficacy and pharmaceutical mechanism of action of AT-II combined with IFN- gamma synergistically against CRC by regulating the NF-kB p65/PD-L1 signaling pathway. Methods: HT29 and HCT15 cells were treated with AT-II and IFN- gamma alone or in combination and cell viability, migration, and invasion were then analyzed using Cell Counting Kit-8 (CCK-8) and Transwell assays, respectively. Furthermore, the underlying mechanism was investigated through western blot assay. The role of AT-II combined with IFN- gamma on tumor growth and lung metastases was estimated in vivo . Finally, the population of lymphocytes in tumor tissues of lung metastatic C57BL/6 mice and the plasma cytokine levels were confirmed by flow cytometry and enzyme-linked immunosorbent assay (ELISA). Results: AT-II or the combination IFN- gamma significantly inhibited the growth and migration abilities of CRC cells in vitro and in vivo . The biological mechanisms behind the beneficial effects of AT-II combined with IFN- gamma were also measured and inhibition of p38 MAPK, FAK, Wnt/ beta -catenin, Smad, and NF-kB p65/PD-L1 pathways was observed. Moreover, AT-II combined with IFN- gamma significantly inhibited HCT15 xenograft tumor growth and lung metastases in C57BL/6 mice, which was accompanied by lymphocyte infiltration into the tumor tissues and inflammatory response inactivation. Conclusions: The results showed that the AT-II in combination with IFN- gamma could be used as a potential strategy for tumor immunotherapy in CRC. More importantly, the mechanism by which AT-II suppressed CRC progressions was by inhibiting the NF-kB p65/PD-L1 signal pathway.