Haizao Yuhu Decoction (HYD), a classic Traditional Chinese Medicine (TCM) formula, is used to treat Yingbing, which corresponds to goiter in modern medicine. Although HYD has been extensively studied for goiter, its specific active components and the underlying mechanisms of action remain to be fully elucidated. This study investigated the effects and mechanisms of HYD in goiter using untargeted LC-MS metabolomics (comparing HYD decoction and HYD-containing serum), network pharmacology for target prediction, and in vitro validation with a TSHR human monoclonal antibody M22 (TSHR hMAb M22)-induced thyroid cell proliferation model, followed by Western blotting to verify key pathway proteins. The comparison of metabolomic profiles between HYD decoction and HYD-containing serum revealed that 2091 metabolites (90.05%) were downregulated, while 231 were upregulated in medicated serum. Network pharmacology predicted targets including AKT1, PIK3CA, ESR2, ESR1, and TP53. HYD-containing serum exhibited pro-apoptotic effects on the cell proliferation model. The underlying mechanism may primarily involve the upregulation of the p53/PUMA/p21 pathway, which subsequently affects caspase-3 activity. Furthermore, the action of HYD may involve non-p53 multiple signaling pathways. HYD may exert its effects by modulating multiple signaling pathways, including p53/PUMA/p21, leading to apoptosis in thyroid cells.

BackgroundIn 2023, cardiovascular diseases caused an estimated 19.2 million deaths worldwide, accounting for approximately one-third of all global deaths. Despite this heavy burden, there is limited evidence comparing the effects of various exercise modalities on functional capacity, arterial stiffness, and lipid profiles in patients with atherosclerotic cardiovascular disease (ASCVD). This network meta-analysis evaluated 12 exercise interventions to guide personalized exercise prescriptions.MethodsA systematic search of PubMed, Embase, the Cochrane Library, and Web of Science was conducted from their inception until June 10, 2026, to identify randomized controlled trials assessing exercise interventions in adults with ASCVD. The primary outcomes measured included the 6-minute walk distance (6MWD), pulse wave velocity (PWV), total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C). A network meta-analysis was conducted, and the ranking of interventions was determined using the surface under the cumulative ranking curve (SUCRA). The protocol was registered with PROSPERO (registration No. CRD420261412964).ResultsA comprehensive analysis incorporating 29 RCTs with a cumulative sample size of 2,496 participants was conducted. The network meta-analysis (NMA) indicated that various exercise modalities exhibited unique benefits for specific health outcomes. Resistance Training (RT) was identified as the most effective intervention for enhancing the 6MWD. Both Qigong combined with Dancing (QD) and Aerobic Exercise (AE) were associated with the most pronounced improvements in PWV. In terms of lipid profile modulation, QD was the most efficacious in reducing total cholesterol levels and increasing HDL-C, whereas RT combined with Blood Flow Restriction (BFRT) was superior in improving LDL-C levels.ConclusionDifferent exercise interventions yield specific benefits in managing ASCVD. This study endorses the concept of “goal-oriented exercise prescription” and suggests that exercise regimens should be meticulously tailored to align with patients’ individualized treatment goals, in conjunction with standard pharmacological therapy.Systematic Review Registrationhttps://www.crd.york.ac.uk/PROSPERO/view/CRD420261412964, identifier CRD420261412964.

BACKGROUND:Skin photoaging is primarily driven by mitochondrial dysfunction. Although natural products have demonstrated protective effects against Ultraviolet (UV)-induced damage, efficient screening strategies for mitochondrial-targeting compounds remain limited. Artificial Intelligence (AI)-assisted high-content screening strategy offers a valuable approach to identify bioactive candidates and elucidate their mechanisms. PURPOSE:This study aimed to employe an AI-assisted high-content screening strategy to identify mitochondrial-enhancing natural compounds that prevent skin photoaging and to evaluate their protective efficacy. METHODS:This study employed an AI-assisted high-content screening strategy to identify mitochondria-targeting compounds that mitigate photodamage. UVB-induced models were established in cells, zebrafish, and mice with UVB lamp irradiation. Mitochondrial morphology and function were quantified using JC-1, TMRM, and MitoTracker probes. Cell viability, SA-β-gal activity, and protein expression were assessed by CCK-8 assay, SA-β-gal staining, and Western blotting, respectively. RESULTS:Using AI-assisted high-content screening, we identified quercetin, spermidine, adenosine, Vitamin K2, and Mirabilis jalapa extract as mitochondrial protective compounds. Two optimized combinations, CC-1 and CC-2, restored ATP production, TCA cycle flux, and mitochondrial morphology in UVB-exposed human dermal fibroblasts (HDFs), reduced SA-β-gal activity, and upregulated COLLAGEN I, LAMIN B1, and SIRT3 expression. Both combinations alleviated the UVB-induced senescence phenotype by upregulating SIRT3. In a UVB-exposed zebrafish tail amputation model, CC-2 markedly promoted fin regeneration. These findings were validated in UVB-irradiated mice, where CC-2 demonstrated superior efficacy. CONCLUSION:Using AI-assisted high-content screening, we formulated two optimized combinations that alleviate UVB-induced skin photoaging, with CC-2 exhibiting superior efficacy, highlighting its potential as cosmetic additive.

ETHNOPHARMACOLOGICAL RELEVANCE:Bawei Chenxiang Wan (BWCX) is a classical Tibetan medicinal formula originating from the canonical Tibetan medical text The Four Tantras. It is traditionally used to activate qi circulation, nourish the heart, tranquilize the mind and enhance intelligence. It has been applied for over a thousand years on the Qinghai-Tibet Plateau to treat cardio-cerebral hypoxic syndromes such as high-altitude (HA) adverse qi flow and mental trance, which are attributed to an imbalance among the three humors (rlung, mkhris-pa, and bad-kan). The traditional applications of BWCX closely align with the clinical manifestations of modern Chronic Mountain Sickness (CMS). However, its effective components and mechanisms of action anti-CMS remain unclear. AIM OF THE STUDY:This study aims to systematically elucidate the material basis of BWCX anti-CMS and its underlying mechanism of cardio-cerebral protection through the regulation of oxidative stress. MATERIALS AND METHODS:Chemical constituents of BWCX were characterized by UHPLC-MS. To identify core therapeutic targets, we employed network pharmacology and analyzed CMS patient transcriptomes (GSE145774, GSE103940) by combining weighted gene co-expression network analysis (WGCNA) with the random forest (RF) algorithm. Molecular docking was used to validate interactions between active components and predicted targets. A physiologically relevant CMS model was established by exposing 60 mice to a natural HA environment (Lhasa, 3650 m) for 8 weeks. Hematological parameters were measured; oxidative stress markers (malondialdehyde [MDA], Catalase [CAT], Total Superoxide Dismutase [T-SOD], glutathione peroxidase [GSH-Px], total antioxidant capacity [T-AOC]) and energy metabolism (adenosine triphosphate [ATP]) in cardiac tissues were assessed by ELISA; and the expression of core targets was analyzed using RT-qPCR and Western blot to verify the regulatory effect of BWCX on the AKT1/FOXO3a/CAT signaling axis, and the binding situation of FOXO3a to the promoter region of CAT was analyzed using ChIP-qPCR to verify the regulatory relationship of FOXO3a on CAT. RESULTS:A total of 78 chemical components were identified, including 18-β-glycyrrhetinic acid, β-asarone and quassin. Integrative bioinformatics analysis pinpointed AKT1, FOXO3 and CAT as core targets of BWCX anti-CMS, with significant enrichment in oxidative stress-related pathways. Molecular docking results showed that the binding energies of costunolide with AKT1, FOXO3a, CAT and ESR2 were -9.5, -8.8, -11.1 and -9.9 kcal/mol, respectively; while those of 18-β-glycyrrhetinic acid with the above targets were -9.1, -8.2, -10.2 and -9.2 kcal/mol, respectively, indicating their strong potential for target binding. Animal experiments demonstrated that BWCX intervention significantly reversed CMS-induced hematological abnormalities [increased red blood cells (RBC), hemoglobin (HGB), hematocrit (HCT) and platelets (PLT)], ameliorated cardiac oxidative stress (reduced MDA; elevated T-AOC, T-SOD, GSH-Px, CAT, ATP), and corrected the dysregulated molecular expression: it suppressed the upregulation of AKT1 mRNA/protein and p-AKT1/AKT1 ratio, while restoring the downregulated mRNA/protein expression of FOXO3a and CAT and the p-FOXO3a/FOXO3a ratio in cardiac and hippocampal tissues (P < 0.05). RT-qPCR analysis of AKT1, FOXO3a, and CAT mRNA expression in vitro revealed trends consistent with those observed in the in vivo experiments. It was also verified that significant enrichment of FOXO3a was detected in the CAT promoter region under hypoxic stress (P < 0.05). FOXO3a binds to the CAT promoter in H9C2 cells, thereby confirming that CAT is a direct transcriptional target of FOXO3a. CONCLUSIONS:This study provides the first evidence that BWCX alleviates CMS-induced cardio-cerebral hypoxic injury by modulating the AKT1/FOXO3a/CAT signaling axis, thereby rebalancing oxidative stress and improving hemorheology. These findings provide a modern pharmacological interpretation of its traditional cardiocerebral protective effect, offer a paradigm for ethnomedicine research modernization, and lay a scientific foundation for its clinical application and quality control.

Age-related cataract (ARC) remains the leading cause of blindness worldwide. Sagittaria sagittifolia polysaccharide (SSP) extract, a key component of Sagittaria sagittifolia L., exhibits anti-oxidant and anti-apoptotic effects with potential applications in ARC. This study aimed to explore the therapeutic potential of SSP in ARC and the underlying mechanisms. In sodium selenite-induced cataracts in rats and hydrogen peroxide (H2O2)-induced human lens epithelial B3 (HLEB3) cells, SSP significantly improved lens opacity and pathological changes and alleviated apoptosis and endoplasmic reticulum stress (ERS)-related injury indicators (by inhibiting the intracellular Ca2+ and protein expression of Bcl-2-associated X, cleaved caspase-3, binding immunoglobulin heavy chain protein, protein kinase RNA-like kinase, inositol-requiring enzyme 1α, activating transcription factor 6, C/EBP homology protein, c-Jun N terminal kinase, caspase-12, and calpain-2). In addition, SSP increased the expression of nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1, sarco/endoplasmic reticulum-type calcium transport ATPase 2, and B-cell lymphoma-2. After applying Nrf2 knockdown technology by transferring short interfering RNA in HLEB3 cells, SSP demonstrated its protective role by activating Nrf2 and inhibiting ERS-mediated apoptosis. These findings indicate that SSP may protect against ARC by regulating Nrf2/ERS-mediated apoptosis, providing potential evidence for its use in preventing or delaying ARC.

Background: Lung cancer has the highest morbidity and mortality of all tumors, and the development of TKI drugs targeting EGFR activating mutations has brought lung cancer treatment into the targeted era. In view of their low efficacy and susceptibility to drug resistance, there is an urgent need to find strategies to increase their efficacy and reduce the incidence of drug resistance. Methods: In this study, we examined the distribution and probability of EGFR mutations in non-small cell lung cancer patients in the cBioPortal database and compared the survival prognosis of patients with normal and abnormal EGFR, NSCLC patients treated with and without TKI, and NSCLC patients with different EGFR gene copy numbers. We established a mouse lung cancer model and examined the histomorphological characteristics of lung tissues via hematoxylin and eosin staining. Additionally, changes in the copy number of the EGFR gene and its protein expression levels were detected using RT-qPCR and Western blotting. Furthermore, we quantified the concentration of the EGFR protein using ELISA. Results: We found no significant advantage of EGFR-TKI therapy over first-line chemotherapeutic agents in patients with EGFR-abnormal NSCLC. The reason for this may be related to the abnormal EGFR gene copy number; the higher the copy number increases, the worse the survival prognosis of the patients. In molecular biology experiments, we demonstrated that ginsenoside Rg3 down-regulated the copy number of 18, 19, 20, and 21 exons and protein expression of EGFR in lung adenocarcinoma cells. The results of in vivo pharmacodynamic assays confirmed that sequential administration of ginsenoside Rg3 with TKI drugs could achieve a gainful complementary effect. Conclusions: Ginsenoside Rg3 down-regulates the copy number of EGFR important exons in EGFR-mutant cells of lung adenocarcinoma and reduces EGFR protein expression, thus providing a high gainful complementary effect in combination with EGFR-TKI.

Background:Extreme humidity exacerbates gastrointestinal disorders by disrupting gut microbiota homeostasis, compromising the intestinal barrier and triggering immune dysregulation. Coix seed (Coix lacryma-jobi L.), widely used in traditional Chinese medicine for eliminating dampness, shows promise; however, its mechanisms require further elucidation. Methods:A Coix seed decoction (CD) was administered to rats exposed to 85% relative humidity (8 h/day) for 30 days. Forty rats were randomly divided into four groups: control, high-humidity exposed, Coix seed-treated control and Coix seed-treated high-humidity. The chemical composition of the CD was characterised using ultra-high-performance liquid chromatography-mass spectrometry (LC-MS), and faecal moisture content, body weight and histopathological changes were assessed. Colonic tissues were analysed by haematoxylin-eosin staining and transmission electron microscopy for structural integrity. Gut microbiota were profiled using 16S ribosomal RNA sequencing, followed by bioinformatic analysis of diversity, differential abundance and co-occurrence networks. Untargeted metabolomics was performed using LC-MS to identify metabolic alterations, and the serum inflammatory cytokines (tumour necrosis factor alpha [TNF-α], interleukin [IL]-6, IL-17) were measured by enzyme-linked immunosorbent assay. Results:High humidity disrupts gut homeostasis in rats by inducing intestinal damage, inflammation, dysbiosis and lipid metabolic disorders. Coix seed decoction significantly alleviated these effects, restoring colonic structure, rebalancing the gut microbiota (eg increasing Erysipelatoclostridium spp. Akkermansia spp. Lactobacillus spp. and reducing Escherichia spp. Shigella spp.) and correcting sphingolipid metabolism (eg sphingomyelin, ceramide). Metabolomic and transcriptomic analyses revealed that CD suppressed immune-related pathways (eg T/B cell receptor signalling, Th1/Th2 differentiation), consistent with reduced serum levels of proinflammatory cytokines (TNF-α, IL-6, IL-17). Correlation networks highlighted interactions between key microbes and metabolites, suggesting a regulatory role for Ruminococcus spp., NK4A214_group spp. and Prevotellaceae NK3B31 spp. in the therapeutic effects of CD. Conclusion:Our findings provide experimental evidence that Coix seed mitigates high humidity-induced gut injury through microbiota remodelling, lipid metabolic regulation and immune modulation.

Objectives: To investigate the therapeutic effects of Qin Gui Wan (QGW) and its disassembled functional drug groups, Wenyang Zhuhuo (WYZH) and Xinwen Zhuyang (XWZY), on letrozole-induced PCOS rats. Methods: PCOS rat model was established by administering letrozole for 21 days. The rats were divided into control, PCOS, Diane-35, QGW, WYZH and XWZY groups. The changes of body weight, ovarian coefficient, estrous cycle and sex hormone levels were observed. The ovarian histological characteristics and ovulation were observed by HE staining. P450arom, SF-1, and AMPK/PGC-1 alpha pathway mRNA and protein expression were analyzed using qRT-PCR, WB, and IHC. The AMPK inhibitor Compound C (CC) was used to explore the treatment mechanism of QGW in granulosa cells. And UHPLC-MS/MS was used to performed chemical composition analysis. Results: QGW, WYZH, and XWZY can correct the disordered estrous cycle of PCOS rats and improve the serum hormone status of rats to varying degrees. HE results indicated that QGW, WYZH, and XWZY improved ovarian polycystic changes and normalized ovulation. qRT-PCR, WB, and IHC results demonstrated that QGW, WYZH, and XWZY increased PGC-1 alpha, SF-1, and P450arom mRNA and protein expression in the ovaries of PCOS rats. The level of AMPK mRNA in the ovaries of QGW and its disassembled prescriptions increased, while only WYZH and XWZY rats showed increased ovarian AMPK levels. CC attenuated the activation of AMPK, PGC-1 alpha, SF-1, and P450arom mRNA by QGW. Conclusions: This study demonstrates that QGW alleviates abnormal oocyte development in PCOS rats, possibly by enhancing P450arom expression via the AMPK/PGC-1 alpha pathway, thus restoring normal androgen-estrogen balance and follicular development.

ETHNOPHARMACOLOGICAL RELEVANCE:Ulcerative colitis (UC) is a chronic, relapsing inflammatory bowel disease primarily characterised by persistent disruption of the colonic epithelial barrier. Despite the proven clinical efficacy of the traditional Chinese medicine formulation Wumei Wan (WMW), its pharmacological mechanisms remain inadequately understood. AIM OF THE STUDY:This study aims to elucidate the therapeutic mechanisms through which WMW promotes the repair of colonic epithelial barrier damage in UC. METHODS:UC was induced in rats via dextran sulfate sodium (DSS), followed by treatment with low- and high-dose WMW or the positive control 5-ASA. Therapeutic effects of WMW on colonic barrier damage were assessed using conventional indices and serum biomarkers. Mechanistic insights were derived through network pharmacology analysis and transcriptome sequencing (RNA-seq), with experimental validation performed using reverse transcription quantitative PCR, Western blot analysis, and immunofluorescence (IF). RESULTS:Both high- and low-dose WMW improved serum biomarkers associated with intestinal barrier function (endotoxin, diamine oxidase, and D-lactate), alongside conventional metrics such as disease activity index, ZO-1, and MUC2 expression. Network pharmacology and RNA-seq analyses identified key therapeutic mechanisms, including modulation of pyroptosis, regeneration of LGR5+ intestinal stem cells (ISCs), and regulation of inflammatory signalling pathways. Experimental validation confirmed that WMW inhibited pyroptosis-associated proteins (NAIP5, NAIP6, NLRC4, GSDMD-N, caspase-1 p20, IL1β, and IL18) while promoting ISC regeneration via upregulation of LGR5, ASCL2, and IL11RA1, with co-localisation of LGR5 and IL11RA1-a novel finding. Additionally, WMW suppressed phosphorylation of JAK2/STAT3 pathway components (p-JAK2 and p-STAT3) and elements of the STING1/IRF3 pathway (p-STING1, p-IRF3, and p-NF-κB p65), offering moderate complementary regulation of inflammatory pathways in UC. CONCLUSIONS:WMW mitigates UC-associated epithelial barrier dysfunction through a multifaceted mechanism involving inhibition of NAIP5/6-NLRC4 pathway-mediated epithelial pyroptosis, enhancement of ASCL2/IL11RA1-dependent LGR5+ ISC regeneration, and suppression of JAK-STAT and STING1/IRF3 inflammatory signalling. This polypharmacological action, driven by WMW's complex phytochemical composition, underscores therapeutic potential of multi-target herbal medicines in addressing the multifactorial pathogenesis of UC.

Activation of the NOD-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome and oxidative stress are key pathological hallmarks of inflammatory bowel disease (IBD) and represent novel targets for therapeutic intervention. The therapeutic potential of the naturally occurring flavonoid kaempferol (KAE) in IBD, along with its underlying molecular mechanisms, remains incompletely understood. We established dextran sulfate sodium (DSS)-induced colitis and lipopolysaccharide (LPS)-induced systemic inflammation mouse models. KAE was administered as an intervention. In vitro, peritoneal macrophages (PMs) were stimulated to activate the NLRP3 inflammasome. We comprehensively evaluated the effects of KAE on interleukin-1β (IL-1β) release, NLRP3 inflammasome assembly, and mitochondrial reactive oxygen species (mt-ROS) production. Additionally, we assessed its impact on the Keap1-Nrf2 pathway. Molecular docking and ubiquitin-dependent degradation assays were conducted to confirm Nrf2 as a direct target of KAE. KAE significantly attenuated colitis development, marked by reduced NLRP3 expression and enhanced Nrf2 activation. It inhibited both the priming and assembly phases of NLRP3 inflammasome activation. Notably, Nrf2 inhibition completely abolished KAE-mediated removal of mt-ROS and downstream suppression of NLRP3 inflammasome activation. Mechanistically, KAE prevented Nrf2 proteolysis by directly binding to Arg415 of Keap1. Our findings demonstrate that KAE acts as an mt-ROS scavenger to protect against inflammatory pyroptosis, offering novel mechanistic insights and a promising therapeutic strategy for colitis treatment.

Rhodiola crenulata(Hook.F.et Thoms.)H.Ohba(Hongjingtian,RC)thrives mainly in high-altitude regions including but not limited to the Himalayan region,Tibetan areas in China,northwest Yunnan,and Western Sichuan.(1,2)The dried root and rhizome of RC have a long history of medicinal use,primarily valued for its adaptogenic effects and significant potential for further development.(3,4)Meanwhile,RC is documented in the Chinese Pharmacopoeia and classical texts on Chinese medicine(CM),such as Sheng Nong's Herbal Classic.

Over the past five decades, anthropogenic activities have precipitated a sustained rise in global temperatures. This warming trend enhances the atmosphere’s water-holding capacity, thereby elevating atmospheric humidity levels. Prolonged exposure to high humidity (HH) adversely impacts nearly all organ systems are increasingly being recognized. As the respiratory system interfaces directly with atmospheric conditions, HH exposure exerts immediate pathological effects on pulmonary structures and functions. The past decades in environmental medicine research have been characterised by a greater understanding of HH to respiratory disease. This Seminar reviews the impact of HH on a spectrum of prevalent respiratory disorders, with an emphasis on respiratory infectious diseases, chronic ventilatory dysfunction, allergic airway diseases, and particulate matter-induced lung injury.

Cigarette smoke (CS), an indoor environmental pollution, is an environmental risk factor for diverse neurological disorders. However, the neurotoxicological effects and mechanisms of CS on Alzheimer's disease (AD) progression remain unclear. We found that CS accelerated the progression of AD, including increasing β-amyloid (Aβ) plaque deposition and exacerbating cognitive decline. Mechanistically, CS exposure increased the levels of NOD-like receptor protein 3 (NLRP3), which impaired autophagic flux in microglia by activating the mammalian target of rapamycin (mTOR) signal. Metabolomics analysis revealed an upregulation of lactate levels and an increase in global protein lysine lactylation in the brain tissue of CS-exposed AD-transgenic mice. Immunoprecipitation-Mass Spectrometry and chromatin immunoprecipitation assays demonstrated that CS elevates H4K12 lactylation (H4K12la) levels, which accumulate at the promoter region of NLRP3, leading to the activation of its transcription. Via inhibiting lactate or NLRP3 activation, oxamate and MCC950 alleviates these CS-induced effects. Therefore, our data suggest that the CS-induced increase in lactate levels triggers NLRP3 transcriptional activation through H4K12la, which subsequently leads to mTOR-mediated autophagy dysfunction in microglia, promoting microglial activation and resulting in Aβ plaque accumulation in AD-transgenic mice. This provides a new mechanism and potential therapeutic target for AD associated with environmental factors.

ETHNOPHARMACOLOGICAL RELEVANCE:Aging is a time-dependent decline in physiological function that reduces quality of life. In traditional Chinese medicine (TCM), aging is viewed as "deficiency" complicated by blood stasis and qi stagnation. "Kang Shuai Lao Pian" (KSLP) is a Ming Dynasty herbal formula widely used to repenish qi, nourish yin, calm the heart, and tranquilize the mind. AIM OF THE STUDY:We aimed to evaluate the systemic anti-aging activity of KSLP and to determine its mode of action in naturally senescent mice. MATERIALS AND METHODS:Male 16-18-month-old mice received daily intragastric gavage of KSLP (450 mg/kg) for 8 weeks. Skin structure, adipose distribution, neurotransmitters, adipose-derived stem cells (ADSCs) morphology and differentiation, 16S rRNA gene sequencing and untargeted metabolomics analysis of cecal feces were assessed. RNA sequencing (RNA-Seq) was performed on senescent ADSCs; metabolic-microbe correlations were analyzed by Pearson correlation. RESULTS:KSLP restored skin structure and dermal collagen density, normalized dopamine, norepinephrine, acetylcholine, and 5-hydroxytryptamine levels, and improved ADSCs osteogenic/adipogenic capacity- changes mechanistically linked to PI3K-Akt and cytokine-cytokine receptor interaction signaling pathways. In gut, KSLP reversed age-associated shifts in Firmicutes, Epsilonbacteraeota, Ruminococcaceae, and Helicobacter and modulated 57 metabolites particularly within amino acid, fatty acid, and nucleotide metabolic pathways. Correlation analysis identified Ruminococcaceae_UCG_014, Candidatus_Saccharimonas, Helicobacter, Rikenellaceae_RC9, and Ileibacterium as keystone taxa linked to KSLP-responsive metabolites. CONCLUSION:KSLP systemically improves skin architecture, ADSCs function, gut-microbiome balance, and metabolic profiles in naturally aged mice, supporting its clinical potential against age-related decline. Bioactivity-guided identification of active compounds and in vivo mechanistic validation are warranted.

Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive fibrotic disease with an unclear etiology and no effective treatment. This study aims to elucidate the pathogenic mechanism networks involving multiple targets and pathways in IPF. Extracts and metabolites of Astragali Radix (AR) and Angelicae Sinensis Radix (ASR), two well-known traditional Chinese medicines, have demonstrated therapeutic effects on IPF. However, the underlying mechanisms of AR and ASR remain unclear. Utilizing network pharmacology analysis, the disease targets associated with IPF were obtained from the GeneCards database and Online Mendelian Inheritance in Man (OMIM) database. Targets of AR and ASR were identified using the TCM Systems Pharmacology Database and Analysis Platform and Swiss Target Prediction. A proteinprotein interaction (PPI) network was subsequently constructed and analyzed using the STRING database and Cytoscape software. Gene ontology enrichment (GO) analysis and kyoto encyclopedia of genes and genomes (KEGG) analysis were conducted using DAVID. Additionally, a component-target-pathway network were employed to identify the main active components, and molecular docking was performed between these components and proteins encoded by key targets. Finally, in vitro studies were conducted based on network pharmacology. A total of 260 common targets between IPF and drug targets were identified and included in the PPI network, in which TNF, IL-6, STAT3, AKT1, VEGFA, SRC, EGFR, INS, JUN, and IL1B were predicted as key targets. These 260 targets were enriched in the PI3K-AKT signaling pathway, HIF-1 signaling pathway, TNF signaling pathway, MAPK signaling pathway, FOXO signaling pathway, and Pathways in cancer. Docking scores ranged from – 4.1 to – 9.5 kcal/mol, indicating a strong binding affinity between the main active compounds and key targets. In vitro studies have indeed shown that Quercetin and Magnolol can alleviate the expression of epithelial-mesenchymal transition in the A549 cells caused by IL-6. The treatment with AR and ASR resulted in a reduction of mRNA levels for key targets HIF-1α and α-SAM. Additionally, the protein expression levels of P-JAK2/ JAK2, P-STAT3/ STAT3, and α-SMA/ β-Actin were also reduced. These results support the therapeutic potential of AR and ASR in ameliorating pulmonary fibrosis and provide insight into the molecular mechanisms involved in their therapeutic effects.

Objective: To explore and validate the potential targets of Paeoniae Radix Alba (P. Radix, Bai Shao) in protecting against chemical liver injury through network pharmacology, molecular docking technology, and in vitro cell experiments. Methods: Network pharmacology was used to identify the common potential targets of P. Radix and chemical liver injury. Molecular docking was used to fit the components, which were subsequently verified in vitro. A cell model of hepatic fibrosis was established by activating hepatic stellate cell (HSC)-LX2 cells with 10 ng/mL transforming growth factor-β1. The cells were exposed to different concentrations of total glucosides of paeony (TGP), the active substance of P. Radix, and then evaluated using the cell counting kit-8 assay, enzyme-linked immunosorbent assay, and western blot. Results: Analysis through network pharmacology revealed 13 key compounds of P. Radix, and the potential targets for preventing chemical liver injury were IL-6, AKT serine/threonine kinase 1, jun proto-oncogene, heat shock protein 90 alpha family class A member 1 (HSP90AA1), peroxisome proliferator activated receptor gamma (PPARG), PTGS2, and CASP3. Gene Ontology (GO) enrichment analysis indicated the involvement of response to drugs, membrane rafts, and peptide binding. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis revealed that the main pathways involved lipid and atherosclerosis and chemical carcinogenesis-receptor activation. Paeoniflorin and albiflorin exhibited strong affinity for HSP90AA1, PTGS2, PPARG, and CASP3. Different concentrations of TGP can inhibit the expression of COL-Ⅰ, COL-Ⅲ, IL-6, TNF-α, IL-1β, HSP-90α, and PTGS2 while increasing the expression of PPAR-γ and CASP3 in activated HSC-LX2 cells. Conclusion: P. Radix primarily can regulate targets such as HSP90AA1, PTGS2, PPARG, CASP3. TGP, the main active compound of P. Radix, protects against chemical liver injury by reducing the inflammatory response, activating apoptotic proteins, and promoting the apoptosis of activated HSCs.

Meroterpenoids in the traditional Chinese medicine Arnebia euchroma are thought to be key components in its anti-inflammatory activity. In the present study, 17 meroterpenoids including four types of structural skeletons (1-17), together with a monoterpenoid (18), were isolated from the roots of A. euchroma. HRESIMS, 1D and 2D NMR, electronic circular dichroism, and quantum computing-assisted methods were employed to determine the structures of four previously undescribed compounds (1-3, and 14). Zicaomeroterin B (2) feature with a novel benzo[b]oxepin moiety was confirmed by X-ray analysis based on the crystalline mate method. Meroterpenoids 2, 5, 6, 13, and 17 exhibited significant inhibition against the production of NO, IL-6, and TNF-α in LPS-stimulated macrophages without obvious cytotoxic effects. Furthermore, compounds 2, 5, and 6 significantly inhibited the phosphorylation activation of NF-κB p65 and its nuclear translocation in luciferase reporter test, immunoblotting, and immunofluorescence imaging, which in turn inhibited the NF-κB pathway and exerted anti-inflammatory effects. These findings suggested that meroterpenoids 2, 5, and 6 in A. euchroma are the potential lead compounds for anti-inflammatory agents based on NF-κB signaling pathway.