Background: miR-27b-3p is dysregulated in many cancers, but its expression, clinical role, and mechanism in pancreatic cancer are not fully understood. Objective: This study aimed to evaluate the clinical value of miR-27b-3p in PC and its role in pancreatic cancer (PC) progression by targeting PPARG. Method: qRT-PCR was used to measure miR-27b-3p and PPARG levels in 106 paired PC tissues and adjacent non-tumor tissues, and in PC cell lines (PANC-1, SW1990, BxPC-3, AsPC-1) and HPDE cells. Kaplan-Meier and multivariate Cox analyses assessed miR-27b-3p prognostic value. CCK-8 and Transwell assays evaluated the impact of miR-27b-3p inhibition on PC cell function. Dual-luciferase reporter and Pearson correlation analyses confirmed the targeting of PPARG by miR-27b-3p. A PPARG knockdown rescue experiment verified that miR-27b-3p function depends on PPARG. Results: miR-27b-3p was highly expressed in PC tissues and cell lines and linked to lymph node metastasis, distant metastasis, elevated CA19-9 levels, and poor overall survival. Multivariate Cox analysis showed high miR-27b-3p expression as an independent predictor of poor prognosis. In vitro, miR-27b-3p inhibition suppressed PANC-1 cell proliferation, migration, and invasion. PPARG was a direct target of miR-27b-3p and was downregulated at both mRNA and protein levels. A negative correlation was found between miR-27b-3p and PPARG mRNA in PC tissues. PPARG knockdown reversed the effects of miR-27b-3p inhibition, confirming functional dependence. Conclusion: miR-27b-3p functions as an oncogenic miRNA in PC by targeting PPARG and may be a prognostic biomarker and therapeutic target.

BACKGROUND:Pancreatic ductal adenocarcinoma (PDAC) is an aggressive digestive malignancy with poor prognosis. Although apoptosis has been regarded as a principal mechanism through which chemotherapeutic agents exert their anti-tumor effects, PDAC cells exhibit marked resistance to apoptotic cell death, highlighting an urgent need for developing alternative therapeutic strategies. Emerging evidence suggests that pyroptosis can provoke anti-tumor immunity and suppress tumor growth. Gasdermin E (GSDME) is critical in pyroptosis by converting caspase 3 (CASP3)-mediated apoptosis into pyroptosis, thereby eliciting robust anti-tumor immunity. METHODS:We applied a hGLuc-hGSDME-PCA system for high-throughput screening of compounds that induce GSDME-mediated pyroptosis, aiming to develop novel therapeutic strategies for PDAC. Cell migration was evaluated by wound healing and Transwell assays, and colony formation assay was used to assess cell proliferative. Pyroptosis was detected by flow cytometry and Hoechst 33342/PI staining, while the activation and expression of CASP3 and GSDME were analyzed by western blotting. RESULTS:Based on this system, we screened 12 traditional Chinese herbal medicines and identified that Rabdosiae Rubescentis Herba (RRH) significantly induced pyroptosis. Notably, we demonstrated for the first time that oridonin and ponicidin, the primary active constituents of RRH, potentially triggered pyropotosis in PANC-1 cells. Further mechanistic investigation demonstrated that oridonin and ponicidin induced pyroptotic cell death via CASP3/GSDME signaling pathway. CONCLUSION:Collectively, our study validated the reliability of hGLuc-hGSDME-PCA screening system and provided the first experimental evidence that oridonin and ponicidin could induce pyroptotic cell death. These findings suggested their potential as novel therapeutic agents for the treatment of PDAC.

Pyroptosis, an inflammatory programmed cell death, is typically initiated by inflammasomes and executed by gasdermin proteins. Gasdermin D (GSDMD)-mediated pyroptotic cell death and downstream inflammation cascades are crucial innate immune mechanisms implicated in infectious and inflammatory diseases, thus GSDMD is regarded as a novel therapeutic target of various diseases. Inhibiting the activation of GSDMD is an attractive strategy to curb pyroptosis and inflammation. To facilitate pharmacological discovery of GSDMD inhibitors, we aimed to develop a novel high-throughput hGLuc-mGSDMD-PCA screening system with coelenterazine as the substrate to analyze the GSDMD N-terminal (GSDMD-N) oligomerization sensitively. The system was based on lipopolysaccharides (LPS) and nigericin (Nig)-induced pyroptosis model, where luciferase signal intensity was correlated with pyroptotic progression and verified by further experiments. Based on this system, 64 natural products were subjected to screening. 17 candidate compounds of them were identified to suppress the cleavage and aggregation of GSDMD-N. Baohuoside I (BI) and andrographolide (AG) possessed the most potent inhibitory effects on pyroptotic cell death. More importantly, in addition to chlorogenic acid, forsythoside A and other active compounds that had been reported to inhibit pyroptotic possess, isochlorogenic acid A (ICGA), epigoitrin (EPI), and calycosin-7-O-glucoside (CG) were also screened out and firstly confirmed to inhibit pyroptosis via regulating NLRP3/ASC/Caspase-1/GSDMD-dependent pyroptotic process in vitro and in vivo. We have established a useful tool to detect GSDMD-mediated pyroptosis, and systematically investigated and optimized usage conditions. The ease of use and applicability in vitro makes the hGLuc-mGSDMD-PCA system an attractive tool to monitor GSDMD-N oligomerization, which is of great significance for the screening of drugs with anti-pyroptotic and inflammatory activity.

This study investigated the mechanism of TGF-β1-induced Nox4 expression in pulmonary fibrosis (PF) and the anti-fibrotic effects of Tanshinone IIA (Tan-IIA). In a bleomycin-induced pulmonary fibrosis mouse model and in TGF-β1-stimulated fibroblasts, Tan-IIA attenuated fibrosis, oxidative stress, and fibroblast activation. Pharmacological inhibition revealed that the JNK/c-Jun and Smad3 pathways cooperatively mediate TGF-β1-induced expression of Nox4 and fibrotic markers (Collagen I/III, α-SMA). Tan-IIA exerted these effects by dually inhibiting the JNK/c-Jun and Smad2/3 pathways, reducing their phosphorylation and nuclear signaling, which consequently suppressed Nox4 transcription and protein expression. The combination of Tan-IIA with JNK or Smad3 inhibitors synergistically enhanced these effects. We identified a tandem c-Jun/Smad binding element in the Nox4 promoter that is critical for TGF-β1 response. Reporter assays and CUT&RUN experiments confirmed that TGF-β1-induced transcriptional activation depends on an intact c-Jun/Smad binding element and recruitment of c-Jun and Smad2/3. Moreover, Tan-IIA inhibited the enrichment of c-Jun and Smad2/3 at the Nox4 promoter. Collectively, our findings demonstrate that a c-Jun/Smad element integrates profibrotic JNK and Smad signaling to drive Nox4 expression. Tan-IIA presents a novel therapeutic strategy for fibrosis by simultaneously targeting these two key pathways, thereby mitigating Nox4-dependent oxidative stress and fibroblast activation.

A manganese porphyrin wrapped DNA dendrimer (Mn-DD) was developed through enzyme-free DNA self-assembly and simple and mild groove binding of porphyrin. The Mn-DD not only possessed plenty of manganese porphyrin to amplify the chemiluminescence (CL) signal, but also can be modified with diverse groups via DNA hybridization. Combined with an immunosensor array, Mn-DD can be utilized for CL immunoassay of multiple mycotoxins as a universal tag. Under optimal conditions, Mn-DD-based CL imaging immunoassay of aflatoxin B1 (AFB1), ochratoxin A (OTA), and zearalenone (ZEN) exhibited broad linear ranges over 4 orders of magnitude and detection limits as low as 0.87, 0.75, and 0.79 pg mL-1, respectively. It was also utilized in the examination of real coix seed samples, yielding reliable results. High sensitivity, as well as simple operation, low reagent dosage, acceptable accuracy and stability showed the tag and the approach broad application prospects in quality control of food and medicine.

Oxidative stress serves as a driving force for myofibroblast activation in pulmonary fibrosis (PF). As a main enzymatic source of reactive oxygen species (ROS), NADPH oxidase 4 (Nox4) plays a critical role in modulating myofibroblast activation, and has thus emerged as a potential therapeutic target for PF. Tanshinone IIA (Tan-IIA), the most abundant fat-soluble component found in the root and rhizome of Salvia miltiorrhiza Bge., has been demonstrated to suppress ROS-mediated myofibroblast activation by inhibiting Nox4, and thereby ameliorating PF. However, the mechanism through which Tan-IIA regulates Nox4 to prevent myofibroblast activation remains unclear. This study aimed to investigate the protective effects of Tan-IIA against myofibroblast activation in PF, and to elucidate the upstream molecular mechanisms involved in Nox4 regulation. Tan-IIA inhibited myofibroblast activation by reducing extracellular matrix deposition in a mouse model of bleomycin-induced PF. Furthermore, Tan-IIA enhanced the expression of Sestrin2 (Sesn2), while concurrently suppressing Nox4 expression. This effect was verified using an in vitro model of transforming growth factor beta 1 (TGF-β1)-stimulated myofibroblast activation. We further demonstrated that Sesn2 was required for Tan-IIA to act against TGF-β1-induced myofibroblast activation by inhibiting Nox4-mediated oxidative stress. Additionally, both in vitro and in vivo studies revealed that Tan-IIA activates AMP-activated protein kinase (AMPK) and inhibits mammalian target of rapamycin (mTOR) via the upregulation of Sesn2. The findings indicate that Tan-IIA suppresses Nox4 by regulating the Sesn2/AMPK/mTOR signaling pathway, which highlights the crucial effect Sesn2 has in modulating Nox4 expression to prevent myofibroblast activation during PF.

This study aimed to investigate the differences in polysaccharides in Astragali Radix (AR) from different sources, as well as before and after honey processing, through the analysis of enzymatic hydrolysates. The hydrolysis product of astragalus polysaccharides (APS) by β-galactosidase was characterized using hydrophilic interaction ultra-high-performance liquid chromatography coupled with time-of-flight mass spectrometry (HILIC-UHPLC-QTOF/MS). The results indicated that all oligosaccharide fragments were composed of hexoses, with sugar groups linked by 1,6-glycosidic bonds. APS enzymatic hydrolysates fingerprints were established using hydrophilic interaction high-performance liquid chromatography with an evaporative light scattering detector (HILIC-HPLC-ELSD) and comprehensively evaluated based on their similarity. Semi-quantitative analysis of each enzymatic hydrolysate was performed, combined with partial least squares discriminant analysis (PLS-DA) and significance testing to further analyze the main differential sugar fragments of AR in different growth modes and before and after honey processing. Glucose and oligosaccharides with degree of polymerization (DP) of 5 and 6 were identified as indicators to distinguish different growth modes. Glucose can serve as an indicator to distinguish raw AR from honey-processed AR (HAR). This study identified quality evaluation indicators of APS based on saccharide mapping, providing a reference for the development of quality standards for AR.

Nur77, a member of the NR4A nuclear receptor family, plays a critical role in tumorigenesis and cancer progression. Ligands activating its nongenomic functions hold therapeutic promise in oncology. Our previous work identified NB1, a 4,4'-bipyridyl cinnamamide derivative, as a novel Nur77 B-site ligand that induces mitochondrial translocation of Nur77 and triggers Nur77/Bcl-2-mediated apoptosis. Through structure-based optimization, we developed NF1 by replacing the 4-hydroxypiperidine group with an N-methylpentane moiety. This modification enhanced the compound's Nur77-binding affinity and stability. NF1 maintains a similar binding mode to NB1, effectively induced apoptosis via the Nur77-Bcl-2 pathway, and demonstrates significant in vivo antitumor efficacy, acceptable pharmacokinetics, and a high safety profile (LD50 > 500 mg/kg). Collectively, these results establish NF1 as a promising candidate for further development in cancer therapy.

Electroanalytical procedures are often closely bound up the gas molecules. However, the detection limitation of some electroanalytical procedures was largely limited by the lower solubility of gas molecules in liquid. To address this problem, a photoelectrochemical enzymatic biosensor with triphasic interface was designed for antibiotics detection. The hydrophobic porous carbon paper (CP) with atomic layer deposition (ALD) Zinc oxide (ZnO) film and Tungsten disulfide (WS2) sheets were used for fixing laccase (Lac) to form Lac/WS2/ZnO/CP, which contacted with analyst solution on one side and exposed to the gas phase directly on the other. Impressively, the catalytic activity of Lac on WS2/ZnO/CP was promoted by adjusting the oxygen concentration from the gas phase and generated significant electrochemical response for sensitively detecting tetracycline (TC), resulting in minimum detection limit of 1.81 fM in the range of 10-200 μM. Additionally, the recovery rate of environmental samples remained at 96.07 %-104.48 %, confirming the reliability and practicality of the prepared biosensors. This strategy provided a potential method to upgrade the optical properties and enzyme activities of biosensor with eventual applications in bioanalysis.

Glioblastoma multiforme (GBM) is an aggressive brain tumor characterized by metabolic plasticity and resistance to therapy. Understanding the mechanisms underlying GBM's adaptability to metabolic stress is crucial for developing effective treatments. This study investigates the role of Brain Protein I3 (BRI3) in regulating lipid metabolism and autophagy in GBM, and its potential as a therapeutic target. We performed integrative bioinformatics analysis using TCGA-GBM and CGGA datasets to identify lipophagy-related gene signatures. BRI3's function was examined through in vitro studies using GBM cell lines and patient-derived samples. Lipid metabolism and autophagy were assessed under normal and oxygen-glucose deprivation (OGD) conditions in BRI3-knockdown and control GBM cells. Bioinformatics analysis revealed a lipophagy-related gene signature associated with poor prognosis in GBM. BRI3 emerged as a key upregulated gene in GBM, correlating with altered lipid homeostasis and enhanced autophagy. In vitro studies demonstrated that BRI3 knockdown led to lipid accumulation, impaired autophagy, reduced proliferation, and increased apoptosis in GBM cells. Under OGD conditions mimicking the tumor microenvironment, BRI3-depleted cells showed compromised lipid mobilization, autophagy induction, and cell survival compared to controls. Our findings suggest BRI3 as a critical regulator of lipophagy in GBM, enhancing tumor cell resilience to metabolic stress. This study provides insights into GBM's metabolic adaptability and identifies BRI3 as a potential therapeutic target for modulating tumor cell survival in the challenging glioblastoma microenvironment.

INTRODUCTION:Glutamine metabolic reprogramming, mediated by glutaminase (GLS), is an important signal during pulmonary fibrosis (PF) progression. Tanshinone IIA (Tan IIA) is a naturally lipophilic diterpene with antioxidant and antifibrotic properties. However, the potential mechanisms of Tan IIA for regulating glutamine metabolic reprogramming are not yet clear. OBJECTIVES:This study aimed was to evaluate the role of Tan IIA in intervening in glutamine metabolic reprogramming to exert anti-PF and to explore the potential new mechanisms of metabolic regulation. METHODS:Fibrotic characteristics was detected via immunofluorescence and western blotting analysis. Cell proliferation was examined with EdU Assay. Cell metabolites were labeled by using stable isotope [U-13C5]-glutamine. By utilizing 100% 13C glutamine tracers and employing network analysis to investigate the activation of metabolic pathways in fibroblasts, as well as evaluating the impact of Tan IIA on these pathways, we accurately quantified the absolute flux of glutaminolysis, proline synthesis, and the TCA cycle pathway using isotopomer network compartmental analysis (INCA), a user-friendly software tool for 13C metabolic flux analysis (13C-MFA). Molecular docking was used for identifying the binding of Tan IIA with target protein. RESULTS:Tan IIA ameliorate TGF-β1-induced myofibroblast proliferation, reduce collagen I and III and α-SMA protein expression in MRC-5 and NIH-3T3 cells. Furthermore, Tan IIA regulate mitochondrial energy metabolism by modulating TGF-β1-stimulated glutamine metabolic reprogramming in NIH-3T3 cells and inhibiting GLS1 expression, which reduced the metabolic flux of glutamine into mitochondria in myofibroblasts, and also targeted inhibited the expression of Δ1-pyrroline-5-carboxylate synthase (P5CS), P5C reductase 1 (PYCR1), and phosphoserine aminotransferase 1 (PSAT1), and reduced proline hydroxylation and blocked the collagen synthesis pathway. CONCLUSION:Tan IIA reverses glutamine metabolic reprogramming, reduces mitochondrial energy expenditure, and inhibits collagen matrix synthesis by modulating potential targets in glutamine metabolism. This novel perspective sheds light on the essential role of glutamine metabolic reprogramming in PF.

BACKGROUND:Activation of myofibroblasts, linked to oxidative stress, emerges as a pivotal role in the progression of pulmonary fibrosis (PF). Our prior research has underscored the therapeutic promise of tanshinone IIA (Tan-IIA) in mitigating PF by enhancing nuclear factor-erythroid 2-related factor 2 (Nrf2) activity. Nevertheless, the molecular basis through which Tan-IIA influences Nrf2 activity has yet to be fully elucidated.METHODS:The influence of Tan-IIA on PF was assessed in vivo and in vitro models. Inhibitors, overexpression plasmids, and small interfering RNA (siRNA) were utilized to probe its underlying mechanism of action in vitro.RESULTS:We demonstrate that Tan-IIA effectively activates the kelch-like ECH-associated protein 1 (Keap1)-Nrf2 antioxidant pathway, which in turn inhibits myofibroblast activation and ameliorates PF. Notably, the stability and nucleo-cytoplasmic shuttling of Nrf2 is shown to be dependent on augmented autophagic flux, which is in alignment with the observation that Tan-IIA induces autophagy. Inhibition of autophagy, conversely, fosters the activation of extracellular matrix (ECM)-producing myofibroblasts. Further, Tan-IIA initiates an autophagy program through the sestrin 2 (Sesn2)-sequestosome 1 (Sqstm1) signaling axis, crucial for protecting Nrf2 from Keap1-mediated degradation. Meanwhile, these findings were corroborated in a murine model of PF.CONCLUSION:Collectively, we observed for the first time that the Sqstm1-Sesn2 axis-mediated autophagic degradation of Keap1 effectively prevents myofibroblast activation and reduces the synthesis of ECM. This autophagy-dependent degradation of Keap1 can be initiated by the Tan-IIA treatment, which solidifies its potential as an Nrf2-modulating agent for PF treatment.

ETHNOPHARMACOLOGICAL RELEVANCE:Based on ancient classics, Danzhi Tiaozhi Decoction has been successfully used to treat nonalcoholic fatty liver disease for decades. However, its therapeutic mechanisms remain unclear.AIM OF THE STUDY:This study aimed to investigate the effects of Danzhi Tiaozhi Decoction (DZTZD) on metabolic-associated fatty liver disease (MAFLD).MATERIALS AND METHODS:First, we identified the active ingredients of DZTZD and their potential targets in the Traditional Chinese Medicine System Pharmacology database. Using the overlapped genes, we selected the key MAFLD-associated genes, then conducted GO and KEGG pathway enrichment analyses. Furthermore, DZTZD was administered orally to rats, and their serum and liver tissues were examined for absorbed compounds using pharmacochemistry. UPLC-Q-Exactive Orbitrap/MS was used to determine the main compounds. Then, we validated the binding association of the key targets with their active compounds with AutoDock Tools and other software. Finally, the predicted hub targets were experimentally validated.RESULTS:We found 254 active compounds in DZTZD corresponding to 208 targets. Sixteen key genes were identified, and the enrichment analysis revealed multiple signaling pathways, including the AGE-RAGE pathway in diabetic complications and the lipid and atherosclerosis signaling pathway. Next, 160 absorbed components and metabolites were characterized in vivo, and 53 absorbed components and metabolites were characterized in liver tissue. Thirteen parent compounds were identified, including coptisine, quercetin, luteolin, and aloe-emodin. The molecular docking data demonstrated the strongest binding between the active compounds and the core proteins. Moreover, the animal experiments showed that DZTZD decreased body weight, liver weight, lipid accumulation, and ALT, AST, CRP, FFA, IL-6, PEPCK, G6P, TG, TC, and LDL-c serum levels, and increased serum HDL-c levels compared to high-fat induced rats. Besides, the RT-PCR and Western blot showed that DZTZD inhibited the SREBP1c and FAS and increased hyperlipidemia-induced CPT-1A levels. In the high-fat group, JNK phosphorylation increased, and AKT protein phosphorylation decreased, while DZTZD reversed these effects.CONCLUSION:Based on the pharmacological network analysis, pharmacochemistry, and experimental validation, DZTZD can potentially improve MAFLD via the JNK/AKT pathway.

The orphan nuclear receptor Nur77 is emerging as an attractive target for cancer therapy, and activating Nur77’s non-genotypic anticancer function has demonstrated strong therapeutic potential. However, few Nur77 site B ligands have been identified as excellent anticancer compounds. There are no co-crystal structures of effective anticancer agents at Nur77 site B, which greatly limits the development of novel Nur77 site B ligands. Moreover, the lack of pharmaceutical ligands restricts Nur77’s therapeutic proof of concept. Herein, we developed a first-in-class Nur77 site B ligand (NB1) that significantly inhibited cancer cells by mediating the Nur77/Bcl-2-related apoptotic effect at mitochondria. The X-ray crystallography suggests that NB1 is bound to the Nur77 site B with a distinct binding mode. Importantly, NB1 showed favorable pharmacokinetic profiles and safety, as evidenced by its good oral bioavailability in rats and lack of mortality, bodyweight loss, and pathological damage at the 512.0 mg/kg dose in mice. Furthermore, oral administration of NB1 demonstrated remarkable in vivo anticancer efficacy in an MDA-MB-231 xenograft model. Together, our work discovers NB1 as a new generation Nur77 ligand that activates the Nur77/Bcl-2 apoptotic pathway with a safe and effective cancer therapeutic potency.

Pancreatic cancer is highly aggressive and lethal, and treatment options for it are limited. Gasdermin E (GSDME) is highly expressed in pancreatic cancer and can induce pyroptosis. In this type of programmed cell death, cells swell and emit large gas bubbles through their plasma membranes. Hence, GSDME induction is potentially an efficacious therapeutic approach against pancreatic cancer. In the present study, we found that the steroidal saponins polyphyllin I (PPI), collettiside III (CCRIS), and paris saponin V (PSV) significantly inhibited PANC-1, AsPC-1, and BxPC-3 cell proliferation. PPI/CCRIS/PSV altered the morphology of PANC-1 cells and induced the release of lactate dehydrogenase (LDH) from them. Therefore, these three constituents caused PANC-1 cells to undergo pyroptosis. This conclusion was confirmed by propidium iodide (PI) staining and flow cytometry assays. The present work also revealed that PPI/CCRIS/PSV induced pyroptosis via GSDME rather than gasdermin D (GSDMD). Whereas PPI/CCRIS/PSV induced caspase-3 to cleave GSDME, it had no such effect on GSDMD. We also established a PANC-1 xenograft tumor model in BALB/c nude mice and administered CCRIS to them as this compound demonstrated the most substantial pyroptotic effect in the in vitro experiments. This treatment significantly inhibited tumor growth in the mice by activating GSDME-dependent pyroptosis. This research demonstrates demonstrate that pyroptosis induction by PPI/CCRIS/PSV has important implications in basic science and clinical medicine. Future investigations should endeavor to determine the benefits and risks associated with the administration of these steroidal saponins as anti-PDAC therapy.

It is discovered that activated caspase-3 tends to induce apoptosis in gasdermin E (GSDME)-deficient cells, but pyroptosis in GSDME-sufficient cells. The high GSDME expression and apoptosis resistance of pancreatic ductal adenocarcinoma (PDAC) cells shed light on another attractive strategy for PDAC treatment by promoting pyroptosis. Here we report a hGLuc-hGSDME-PCA system for high-throughput screening of potential GSDME activators against PDAC. This screening system neatly quantifies the oligomerization of GSDME-N to characterize whether pyroptosis occurs under the stimulation of chemotherapy drugs. Based on this system, ponatinib and perifosine are screened out from the FDA-approved anti-cancer drug library containing 106 compounds. Concretely, they exhibit the most potent luminescent activity and cause drastic pyroptosis in PDAC cells. Further, we demonstrate that perifosine suppresses pancreatic cancer by promoting pyroptosis via caspase-3/GSDME pathway both in vitro and in vivo. Collectively, this study reveals the great significance of hGLuc-hGSDME-PCA in identifying compounds triggering GSDME-dependent pyroptosis and developing promising therapeutic agents for PDAC.

Evidence indicates that metabolic reprogramming characterized by the changes in cellular metabolic patterns contributes to the pathogenesis of pulmonary fibrosis (PF). It is considered as a promising therapeutic target anti-PF. The well-documented against PF properties of Tanshinone IIA (Tan IIA) have been primarily attributed to its antioxidant and anti-inflammatory potency. Emerging evidence suggests that Tan IIA may target energy metabolism pathways, including glycolysis and tricarboxylic acid (TCA) cycle. However, the detailed and advanced mechanisms underlying the anti-PF activities remain obscure. In this study, we applied [U-13C]-glucose metabolic flux analysis (MFA) to examine metabolism flux disruption and modulation nodes of Tan IIA in PF. We identified that Tan IIA inhibited the glycolysis and TCA flux, thereby suppressing the production of transforming growth factor-β1 (TGF-β1)-dependent extracellular matrix and the differentiation and proliferation of myofibroblasts in vitro. We further revealed that Tan IIA inhibited the expression of key metabolic enzyme hexokinase 2 (HK2) by inhibiting phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR)/hypoxia-inducible factor 1α (HIF-1α) pathway activities, which decreased the accumulation of abnormal metabolites. Notably, we demonstrated that Tan IIA inhibited ATP citrate lyase (ACLY) activity, which reduced the collagen synthesis pathway caused by cytosol citrate consumption. Further, these results were validated in a mouse model of bleomycin-induced PF. This study was novel in exploring the mechanism of the occurrence and development of Tan IIA in treating PF using 13C-MFA technology. It provided a novel understanding of the mechanism of Tan IIA against PF from the perspective of metabolic reprogramming.

Background and objectives: Asiaticoside reduces inflammatory reactions and oxidative stress, the primary causes of photoaging.The authors speculated that asiaticoside might contain therapeutic potential for photoaging.Methods: Network pharmacology and molecular docking were used to explore the mechanisms of asiaticoside in treating photoaging.After achieving the targets of asiaticoside using PharmMapper, SwissTargetPrediction, CTD and BATMAN databases, as well as, targets of photoaging using GeneCards database, the co-targets interaction network was formed and a network of asiaticoside, photoaging, and common targets were constructed by Cytoscape.Next, the common targets were analyzed using GO and KEGG enrichment. Results:The analysis highlighted 202 core targets of asiaticoside were involved in the pathogenesis of photoaging.KEGG indicated asiaticoside performed an anti-photoaging effect through inflammation-and apoptosis-related signalling pathways, especially the PI3K-AKT and NF-κB pathways.Furthermore, the anti-photoaging effect of asiaticoside was verified by human dermal fibroblasts with UVA irradiation in vitro. Conclusion:Asiaticoside may alleviate UVA-induced cell proliferation inhibition, reverse the abnormal gene expressions involved in the PI3K-AKT and NF-κB pathways, and have a high affinity with those core targets.

Pancreatic cancer is highly aggressive and lethal, and treatment options for it are limited. Gasdermin E (GSDME) is highly expressed in pancreatic cancer and can induce pyroptosis. In this type of programmed cell death, cells swell and emit large gas bubbles through their plasma membranes. Hence, GSDME induction is potentially an efficacious therapeutic approach against pancreatic cancer.In the present study, we found that the steroidal saponins polyphyllin I (PPI), collettiside III (CCRIS), and paris saponin V (PSV) significantly inhibited PANC-1, AsPC-1, and BxPC-3 cell proliferation. PPI/ CCRIS/PSV altered the morphology of PANC-1 cells and induced the release of lactate dehydrogenase (LDH) from them. Therefore, these three constituents caused PANC-1 cells to undergo pyroptosis. This conclusion was confirmed by propidium iodide (PI) staining and flow cytometry assays. The present work also revealed that PPI/CCRIS/PSV induced pyroptosis via GSDME rather than gasdermin D (GSDMD). Whereas PPI/CCRIS/PSV induced caspase-3 to cleave GSDME, it had no such effect on GSDMD. We also established a PANC-1 xenograft tumor model in BALB/c nude mice and administered CCRIS to them as this compound demonstrated the most substantial pyroptotic effect in the in vitro experiments.This treatment significantly inhibited tumor growth in the mice by activating GSDME-dependent pyroptosis. This research demonstrates demonstrate that pyroptosis induction by PPI/CCRIS/PSV has important implications in basic science and clinical medicine. Future investigations should endeavor to determine the benefits and risks associated with the administration of these steroidal saponins as anti-PDAC therapy.& COPY; 2023 Elsevier Inc. All rights reserved.