Ficus hirta Vahl (F. hirta) is a traditional medicinal plant used for over 300 years in Chinese ethnic medical systems, including the Yao, Zhuang, and Dai. To provide a rigorous and balanced synthesis, this review conducted a systematic literature search across PubMed, Scopus, CNKI, and Web of Science, focusing on original research regarding the plant's phytochemistry and pharmacology. The extract contains diverse bioactive compounds, including flavonoids, phenolic acids, terpenoids, polysaccharides, and phenylpropanoids, which exhibit significant antibacterial and anti-inflammatory activities. This review summarizes their mechanisms of action: antibacterial effects are achieved through cell wall disruption, metabolic inhibition, and biofilm suppression; while anti-inflammatory effects involve modulating inflammatory mediators, enhancing antioxidant capacity, and regulating immune cells. Furthermore, the synergistic network between these properties and clinical effects—such as digestion promotion and cough relief—is explored. These findings provide a theoretical foundation for the medicinal development of F. hirta and highlight its potential in novel drug discovery.

Candida auris, an emerging multidrug-resistant fungal pathogen, poses a severe global public health threat owing to its high nosocomial transmissibility, considerable mortality, and widespread antifungal resistance. Baicalein (BE), a major bioactive constituent of Scutellaria baicalensis Georgi, exhibits notable antifungal potential, yet its specific molecular mechanisms against C. auris remain poorly elucidated. In this study, we determined the antifungal activity of BE against multiple C. auris isolates, assessed its effects on fungal growth, virulence attributes, adhesion, and biofilm development, validated its in vivo protective efficacy in a Galleria mellonella infection model, and explored the underlying mechanisms via transcriptomic sequencing. BE exerted potent and consistent antifungal activity against all tested strains, with a minimum inhibitory concentration of 1 μg/mL, minimum fungicidal concentrations of 4-8 μg/mL, and a sessile minimum inhibitory concentration of 32 μg/mL. It time- and concentration-dependently suppressed fungal growth, virulence factor expression, adhesion to biological and non-biological surfaces, and biofilm formation, while conferring significant in vivo protection against C. auris infection. Transcriptomic analysis identified 11 adhesion- and biofilm-associated target genes, among which SCF1 and ALS1 served as pivotal regulators. Collectively, BE exerts robust anti-C. auris effects by modulating key target gene expression to interfere with multiple virulence-related processes, providing experimental support for its development as a novel antifungal agent for C. auris infection treatment.IMPORTANCEThe emerging multidrug-resistant fungal pathogen Candida auris has become a critical global public health concern. Its pronounced nosocomial transmissibility, high infection-associated mortality, and extensive cross-resistance to mainstream antifungal agents have created substantial unmet needs in clinical treatment and nosocomial infection control. In this study, we systematically validated the in vitro and in vivo antifungal activity of baicalein against C. auris and elucidated the molecular mechanism underlying its modulation of virulence-related genes. Our findings provide a pivotal experimental basis for the development of novel antifungal therapeutics targeting C. auris infections.

Pseudomonas aeruginosa, which is one of the most common opportunistic pathogenic bacteria, poses severe clinical risks for individuals with compromised immune systems, particularly concerning lung infections. Sodium houttuyfonate (SH), an active constituent isolated from Houttuynia cordata, exhibits limited direct antibacterial efficacy in vitro yet demonstrates notable therapeutic effects against bacterial infections in vivo. Nevertheless, the precise mechanisms underlying in vivo antibacterial pharmacological activity of SH remain unclear. Thus, here we investigate the mechanism by which SH alleviates P. aeruginosa-induced acute pulmonary infection, focusing on its influence on macrophage polarization signaling pathways. First, our findings demonstrate that SH effectively alleviated P. aeruginosa-induced acute pulmonary infection in mice, as evidenced by reduced inflammatory infiltration and alveolar damage in vivo. The results indicate that SH significantly modulated the expression of inflammatory mediators (IL-6, TNF-α, IL-1β, IL-10, TGF-β, Arg-1) and key signaling molecules (NF-κB, TLR4, STAT6, p38MAPK). In vitro, 24-h SH treatment decreased NO production and attenuated macrophage phagocytosis, while shifting cytokine profiles from M1 to M2 phenotypes. Immunofluorescence and flow cytometry confirmed decreased CD86 (M1 marker) and increased CD206 (M2 marker) expression, indicating enhanced M2 polarization. Mechanistically, SH suppressed the TLR-4/MyD88/NF-κB pathway while activating the p38MAPK/STAT6 axis. Genetic manipulation further verified that SH regulates TLR-4 and p38MAPK, thereby controlling downstream signaling and inflammatory responses to combat infection. In conclusion, our study suggests that SH promotes macrophage M2 polarization and reduces excessive inflammation in late-stage P. aeruginosa-induced acute pulmonary infection by modulating macrophage polarization through the suppression of pro-inflammatory signaling via the TLR4/MyD88/NF-κB pathway and activation of the p38 MAPK/STAT6 pathway.

ETHNOPHARMACOLOGICAL RELEVANCE:Oropharyngeal candidiasis (OPC), primarily induced by Candida albicans, poses a significant threat to oral health due to the hypoxic microenvironment established by C. albicans biofilms. Coptis chinensis Franch., a traditional Chinese medicine first documented in Shennong's Classic of the materia medica, has been traditionally employed in the treatment of infectious diseases, including OPC. The ethanolic extract of C. chinensis Franch. (EECF), which is a compound obtained through ethanol extraction from the herb, demonstrates significant therapeutic potential against fungal oral mucosal infections, such as those caused by Candida albicans. However, the possible effects of EECF on hypoxic microenvironment which is key to OPC are unclear. Thus, we hypothesized that EECF may alleviate the hypoxic microenvironment induced by C. albicans infection and modulate the host hypoxia-induced inflammatory response, thereby contributing to the treatment of OPC. PURPOSE:We aim to investigate the potential mechanism of action of EECF for the treatment of OPC by exploring the effects of EECF on the hypoxic microenvironment generated by the C. albicans biofilm and the hypoxia inducible factor-1α (HIF-1α) and activation of immune pathways of host. MATERIALS AND METHODS:We investigated EECF's inhibitory effects on C. albicans biofilm formation using Karl Koffler fluorescent whitener staining, XTT assay, and quantitative real-time polymerase chain reaction (qRT-PCR). In a murine OPC model, therapeutic efficacy was evaluated through hematoxylin-eosin staining, Schiff's periodinium staining, and fungal load quantification. Epithelial barrier protection was assessed via Transwell assays measuring KB cell layer penetration. Mechanistic studies examined EECF's impact on HIF-1α/c-Fos/MKP1/IL-17 pathways using qRT-PCR, western blotting, and immunofluorescence. Finally, gene silencing/overexpression experiments verified the effects of HIF-1α modulation by EECF. RESULTS:In vitro studies revealed EECF inhibits C. albicans hyphal growth, biofilm formation, and cyclic adenosine monophosphate (cAMP) production under both normoxic and hypoxic conditions, accompanied by downregulation of CYR1 and EFG1 in the cAMP pathway. In a murine OPC model, EECF treatment reduced oral fungal burden, repaired tongue papillary architecture, protected mucosal integrity, and suppressed hypoxic microenvironment formation in tongue tissues while modulating HIF-1α/c-Fos/MKP1/IL-17 axis-related genes/proteins. In vitro cellular assays demonstrated EECF impaired C. albicans penetration of KB cell barriers and attenuated infection-induced hypoxia through inhibition of HIF-1α/c-Fos/MKP1/IL-17 pathway components. Hypoxic cellular experiments confirmed EECF's inhibitory effects on these pathways were maintained under low-oxygen conditions. Functional gene validation confirmed EECF regulates the HIF-1α/IL-17 axis through modulation of HIF-1α gene expression. CONCLUSION:Our findings demonstrate that EECF suppresses C. albicans biofilm formation by inhibiting fungal cAMP signaling, which attenuates hypoxia-induced HIF-1α/IL-17 activation in oral epithelial cells, leading to reduced fungal burden and inflammatory pathology. Thus, our results support potential of EECF as a potent therapeutic agent for OPC, and warrant further investigation into its clinical applicability as a precision treatment for fungal-host interaction dysregulation.

The escalating challenge of antifungal resistance in Aspergillus fumigatus underscores the critical need for novel therapeutic combinations. This study identifies the natural naphthoquinone shikonin (SHK) as a potent synergist of amphotericin B (AMB) against A. fumigatus. Checkerboard assays revealed a significant synergistic interaction (FICI <= 0.5), reducing the AMB MIC by up to 8-fold. Mechanistic investigations demonstrated that SHK exerts a multi-target effect: it effectively inhibites biofilm formation and disrupts key components of cell wall biosynthesis. Crucially, real-time imaging and propidium iodide uptake assays confirmed rapid and profound perturbation of membrane integrity. Molecular dynamics simulations provided a structural basis for this effect, indicating that SHK interacts directly with membrane ergosterol, promoting its disorganization and compromising membrane homeostasis. In vivo experiments showed that the combination of SHK and AMB could significantly improve the survival rate (65.2%) compared with any single drug. Our findings elucidate a unique multimodal mechanism by which SHK potentiates AMB, positioning it as a promising adjunctive therapy to combat AMB resistance and improve outcomes in invasive fungal infections.

ETHNOPHARMACOLOGICAL RELEVANCE:Infection of vaginal epithelial cells by Candida albicans (C. albicans) can cause vulvovaginal candidiasis (VVC). Pulsatilla decoction has been reported and used to treat VVC, but the specific mechanism of its protective effect on cells is still unclear. AIM OF THE STUDY:In this study, transcriptomic profiling and molecular docking were used to predict the protective effects and regulatory mechanisms of the n-butanol extract of Pulsatilla Decoction (BEPD) in vaginal epithelial cells stimulated with C. albicans hyphae, and these findings were further validated experimentally. MATERIALS AND METHODS:To prepare the n-butanol extract from Pulsatilla decoction, transcriptome analysis was performed to identify significantly enriched genes and signaling pathways in vaginal epithelial cells stimulated with C. albicans hyphae. RT-qPCR was used to detect the expression levels of genes related to the HIF-1α pathway. Molecular docking was employed to verify the interactions between the main active components of BEPD and HIF-1α, as well as glucose metabolism-related proteins within the pathway. Immunofluorescence was used to detect the expression levels of HIF-1α, PFK2, ENO1, PDK1, Glut1, and LDHA. The cellular contents of ATP, lactic acid, and LDH were measured to reflect changes in sugar metabolism-related products. Western blot and ELISA were used to determine the levels of inflammatory factors such as IL-1β and IL-2 in vaginal epithelial cells stimulated by C. albicans hyphae and treated with BEPD. RESULTS:BEPD exhibits a significant protective effect on vaginal epithelial cells stimulated by C. albicans hyphae. Transcriptomic analysis, including GO enrichment and KEGG pathway analysis, indicates that BEPD can regulate HIF-1α and its associated glucose metabolism disturbances in vaginal epithelial cells exposed to C. albicans hyphae. RT-qPCR confirmed the upregulation of HIF-1α gene expression in cells following BEPD treatment, while promoting the downregulation of genes related to anaerobic metabolism and attenuating the downregulation of genes involved in aerobic metabolism. Furthermore, studies on the binding interactions between drug compounds and cellular proteins revealed that the primary active components of BEPD can specifically bind to HIF-1α and proteins related to glucose metabolism. Immunofluorescence and Western blot analyses of HIF-1α and glucose metabolism-related proteins demonstrated that BEPD treatment results in significant upregulation of HIF-1α protein expression, which, in turn, markedly reduces the expression of genes associated with anaerobic metabolism and proteins involved in aerobic metabolism. ELISA assays further showed that BEPD can protect vaginal epithelial cells stimulated by C. albicans hyphae and decrease the expression levels of inflammatory factors in these cells. CONCLUSIONS:This study demonstrates that BEPD exerts a protective effect on vaginal epithelial cells stimulated by C. albicans hyphae. The underlying mechanism may involve the regulation of HIF-1α signaling and glucose metabolism homeostasis, which represents a novel mechanism against C. albicans infection. These findings provide a new theoretical basis for the prevention and treatment of vulvovaginal candidiasis, and further studies in animal models and clinical settings are warranted in the future.

Background: Plasma fibrinogen (FIB) levels exhibit a significant elevation during the acute phase of ischemic stroke (IS), and their dynamic fluctuations serve as important biomarkers for stroke onset, disease progression, and long-term prognosis. Tong-Qiao-Huo-Xue Decoction (TQHXD) is highly effective in treating blood stasis syndromes affecting the head and face. Nevertheless, the association between TQHXD and FIB in the underlying mechanism of treating IS warrants further investigation. Methods: Proteomics analysis predicted the potential therapeutic targets of TQHXD for IS. An in vivo model of middle cerebral artery occlusion followed by reperfusion (MCAO/R) was created in mice. To explore the interaction between FIB and NLRP3, as well as to verify the particular healing outcomes of TQHXD. Results: An increased blood-brain barrier (BBB) permeability was observed after MCAO/R, accompanied by substantial accumulation of FIB in the brain. In vivo experiments demonstrated that FIB triggered the activation of the NLRP3 inflammasome in microglia. Proteomic analysis revealed a significant increase in FIB levels following model induction, which were markedly reduced after treatment with TQHXD; KEGG pathway enrichment analysis indicated that these changes were primarily associated with the NOD-like receptor signaling pathway. Laser speckle contrast imaging showed that TQHXD treatment significantly improved cerebral blood flow and attenuated brain injury in mice. Fluorescence imaging, ELISA, and Western blotting results collectively demonstrated that TQHXD effectively reduced FIB accumulation and suppressed NLRP3 inflammasome activation. MD and pull-down experiments further demonstrated a strong interaction strength between FIB and NLRP3. Conclusions: FIB accumulates in the ischemic penumbra following CIRI, while TQHXD can effectively down-regulate FIB expression and inhibit NLRP3 inflammasome activation to mitigate CIRI. These findings provide a novel theoretical foundation and treatment direction for stroke management in clinical settings.

ETHNOPHARMACOLOGICAL RELEVANCE:Qi-Huang Decoction (QHD), a combined prescription of Buzhong Yiqi decoction and Dachengqi decoction, is used for gastrointestinal protection post-operation at present. Previously, we found that QHD could ameliorate Candida colitis through Dectin-1 associated pathway, but was rather weak against Candida albicans in vitro. AIM OF THE STUDY:This study explores the protection mechanism of QHD by which miR-32-5p mediated Dectin-1/NF-κB/IκBζ signaling pathway in candidaemia associated renal injury caused by C. albicans. MATERIALS AND METHODS:Signaling pathways were screened by network pharmacology. A murine systemic candidiasis model was established to investigate the therapeutic effect of QHD at 10, 20 and 40 g/kg on candidaemia. The antifungal and anti-inflammatory activity of QHD drug-containing serum (QHD-DS) at 5 and 20 g/kg was monitored in RAW264.7 and MPC5 cells. Dectin-1 inhibitor laminarin (LAM), miR-32-5p mimic and inhibitor, NF-κB antagonist TPCA, and silenced NFKBIZ were employed to analyze the activation of Dectin-1/NF-κB/IκBζ signaling pathway in the protection of QHD against colonization of and inflammation caused by C. albicans. In addition, the antifungal potential of NFKBIZ in combination with miR-32-5p, Dectin-1 and NF-κB was also examined. RESULTS:The NF-κB pathway is most likely to participate in the treatment of QHD against candidaemia. QHD at 40 g/kg significantly prolonged survival rate, ameliorated severity of kidney injury, reduced tissue fungal burden, and neutralized renal inflammation in mice with candidaemia. Similarly, QHD-DS at 5 and 20 g/kg was effective to inhibit C. albicans growth and increased inflammation induced by fungal β-glucan in RAW264.7 and MPC5 cells. QHD deactivated Dectin-1/NF-κB signaling, but LAM abolished this protection. In contrast to downregulated miR-32-5p, overexpressed miR-32-5p assisted QHD-DS at 5 and 20 g/kg to improve candidaemia. NFKBIZ, miR-32-5p target gene, was positively correlated with Dectin-1 and NF-κB. Silenced NFKBIZ abandoned the antifungal and anti-inflammatory potentials of QHD by downregulating TNF-α and IL-1β. Furthermore, si-NFKBIZ could be synergistic with miR-32-5p inhibitor, LAM and TPCA to weaken the QHD-DS's anti-colonization of C. albicans. NFKBIZ, not NF-κB, might be the principle governor in renal inflammation and fungal propagation during candidaemia. CONCLUSION:QHD can ameliorate candidaemia associated renal inflammation via miR-32-5p mediated Dectin-1/NF-κB/IκBζ signaling pathway. QHD presents promising potential for clinical therapy of candidaemia.

OBJECTIVES:To examine the protective effects of cerebrospinal fluid from Tongqiao Huoxue Decoction-treated rats (TQHXD-CSF) against oxygen and glucose deprivation and reoxygenation (OGD/R)-induced injury in murine BV-2 microglial cells and co-cultured HT22 cells. METHODS:In a Transwell co-culture system of BV-2 and HT22 cells, OGD/R+fibrinogen (FIB) injury was induced in BV-2 cells followed by treatment with TQHXD-CSF intervention, and HT22 cells in the lower chamber were cultured under normal conditions. The cells were observed for changes in cell morphology, viability, intracellular ROS level, apoptosis, M1/M2 polarization, and FIB-containing extracellular vesicles (EVs). The cellular expressions of NLRP3, ASC, caspase-1, GSDMD, IL-1β, and IL-18 were quantified using Western blotting, and FIB-NLRP3 binding was confirmed by pull-down assay. RESULTS:OGD/R+FIB injury caused polarization of BV-2 cells to the pro-inflammatory M1 phenotype, increased CD86 expression and release of FIB-containing EVs, and activated the NLRP3 inflammasome. The co-cultured HT22 cells showed reduced cell viability, elevated ROS, and increased cells apoptosis. Treatment with TQHXD-CSF promoted M2 polarization in BV-2 cells, upregulated CD206 expression, suppressed FIB+ EVs secretion, and inhibited NLRP3 inflammasome activation in HT22 cells, which showed significantly lowered expressions of NLRP3, ASC, caspase-1, GSDMD, IL-1β, and IL-18 proteins and hence reduced inflammatory injury and cell apoptosis. Pull-down assay confirmed direct FIB and NLRP3 binding. CONCLUSIONS:In the co-culture system of BV-2 cells and HT22 cells, TQHXD-CSF treatment protects HT22 cells against OGD/R+FIB-induced injury by inhibiting FIB-containing EVs release from BV-2 cells and suppressing NLRP3 inflammasome activation.

Background/Objectives: Xu Chunfu’s Modified Xianglian Pill (XXLP) has been used for centuries in Chinese medicine to treat “diarrhea” and “dysentery,” conditions analogous to modern ulcerative colitis (UC). However, the scientific basis for its efficacy and mechanisms remains unclear. Methods: The chemical composition of XXLP was analyzed via UPLC-ESI-MS/MS. A colitis mouse model was established using DSS, and the therapeutic effects were assessed based on body weight, disease activity index (DAI), colon length, and histopathology. Inflammatory cytokines were measured using ELISA. Proteomic analysis and molecular docking identified key targets, which were validated using LPS-induced HT-29 cells via Western blot (WB), qRT-PCR, immunofluorescence (IF), and transmission electron microscopy (TEM). Gut microbiota composition was analyzed using 16S rRNA gene sequencing. Results: Analysis of XXLP led to the detection of 373 compounds. XXLP significantly improved colitis symptoms, including weight loss and colon shortening, and reduced the concentrations of inflammatory markers IL-1β, IL-18, TNF-α, and IL-6. Proteomics and molecular docking identified NADPH oxidase 2 (NOX2) as a key target of XXLP intervention in mice with colitis. qRT-PCR, WB, IF, and TEM results further confirmed that XXLP effectively suppressed the expression of NOX2 and its associated protein levels. Sequencing analysis of 16S rRNA showed that XXLP significantly increased the relative abundance of beneficial bacterial genera (Muribaculaceae and Ruminococcaceae) while markedly reducing the levels of harmful bacteria (Enterobacteriaceae). Correlation analysis revealed that specific microorganisms were correlated with NOX2-related protein expression and severity of colonic inflammation. Conclusions: XXLP effectively alleviates colitis by suppressing inflammatory responses. Its mechanism involves regulating the NOX2/ROS/mitochondria/NLRP3 axis and altering gut microbiota composition, providing novel insights for colitis treatment.

Sepsis is a life-threatening multi-organ involved diseases due to malfunctional immune response to pathogenic infections. Gut is usually deemed as a motor driver of sepsis. Yi-Qi-Tong-Fu Decoction (YQTFD) is specialized in treating gastrointestinal dysfunction. This study intends to illuminate the therapeutic mechanism of YQTFD by which IL-17 mediated PI3K/AKT signaling pathway regulates sepsis induced intestinal failure via epithelial apoptosis. The potential target signaling pathway was screened with network pharmacology and in silico analysis. The effects of YQTFD at low, medium and high doses (i.e. 5.18, 10.36 and 20.72 g/kg) on murine survival, intestinal pathological change, the gene and protein expressions related to tight junction, inflammation, apoptosis, and the activity of IL-17 mediated PI3K/AKT signaling pathway were assessed in a cecal ligation and puncture (CLP) induced sepsis model. The therapeutic effects of YQTFD drug-containing serum (YQTFD-DS) were also monitored in primary intestinal epithelial and Caco2 cells In silico analysis showed that IL-17 mediated PI3K/AKT signaling pathway might be a potential hub. In CLP mice, YQTFD could significantly extend life span, improve intestinal integrity, and reduce intestinal pathology, inflammation and apoptosis. In Caco2 cells, YQTFD-DS protected against LPS, CM and rIL-17 A induced cellular injuries and apoptosis in a dose-dependent manner. Silenced IL-17RA and PI3K remarkably blocked the therapeutic effect of YQTFD-DS via reactivating IL-17 mediated PI3K/AKT signaling pathway and apoptosis. Z-VAD-FMK also significantly negated the amelioration of YQTFD-DS against apoptosis. YQTFD has satisfactory therapy against sepsis induced intestinal injury via IL-17 mediated PI3K/AKT signaling pathway mediated epithelial apoptosis.

BACKGROUND:Ulcerative colitis (UC) is a chronic inflammatory disease of the gastrointestinal tract. Candida albicans, a common commensal fungus in the human gut, has been increasingly implicated in UC pathogenesis. Qi-Huang decoction (QHD), a traditional Chinese herbal formula known for its spleen-invigorating and purgative properties, is commonly used to restore gastrointestinal function. PURPOSE:This study investigates the therapeutic potential of QHD in treating colitis exacerbated by C. albicans and explores the underlying mechanisms of action. METHODS:A mouse model of colitis was induced using dextran sulfate sodium combined with gavage of C. albicans. Following QHD treatment, colitis severity was evaluated by measuring survival rate, body weight, disease activity index, colon length, and fungal burden, and through histopathological analysis using hematoxylin-eosin staining. The expression of proinflammatory genes IL-1β and TNF-α was quantified, alongside protein levels of key molecules involved in Dectin-1 signaling, including Syk, CARD-9, NLRP-3, Raf-1, and NF-κB. Barrier integrity markers, such as Occludin and Claudin-1, were also examined. To further elucidate QHD's mechanisms, Dectin-1 was inhibited using laminarin. In vitro experiments assessed QHD's antifungal activity against three Candida strains through microdilution, spot assays, and time-kill tests. RAW 264.7 macrophages were employed to study the exposure of fungal cell wall β-glucan and subsequent phagocytosis. Molecular docking simulations predicted interactions between QHD's active compounds and the Dectin-1 receptor. RESULTS:QHD significantly mitigated colitis severity and reduced fungal burden in vivo. QHD enhanced β-glucan exposure on the fungal cell wall, thereby stimulating phagocytosis by RAW264.7 macrophages. QHD effectively activated Dectin-1-mediated signaling pathways and increased proinflammatory levels in RAW 264.7 cells. In colitis mice, QHD treatment markedly reduced inflammation and Dectin-1 signaling following fungal clearance. However, Dectin-1 inhibition with LAM neutralized QHD's therapeutic effects, highlighting the pathway's importance in mediating QHD's efficacy. Interestingly, QHD alone elevated Dectin-1, NF-κB, TGF-β, and IL-10 levels, whereas reduced IL-1β and TNF-α expression, suggesting a dual modulatory role in inflammation. Molecular docking confirmed a potential direct interaction between QHD's bioactive components and the Dectin-1 receptor. CONCLUSION:QHD demonstrates promising therapeutic potential for managing Candida colitis by modulating immune responses and targeting Dectin-1 signaling pathways in clinical settings.

Berberine (BER), a natural isoquinoline alkaloid, exhibits broad-spectrum antifungal activity, yet its mechanism against Aspergillus fumigatus─a leading cause of invasive fungal infections─remains poorly understood. Here, we aim to unveil the mechanism of BER against the pathogenicity of A. fumigatus through mitochondrial dynamics and related pathways. In vitro assays revealed that berberine treatment triggered mitochondrial fragmentation, resulting in reactive oxygen species (ROS) overaccumulation. Subsequent proteomic analyses identified Hog1-MAPK as the central signaling hub activated by ROS stress. Upon activation, Hog1 localizes to the nucleus. ROS scavenging (N-acetylcysteine (NAC) treatment) abolished BER's antifungal effects, confirming the ROS-Hog1-cell cycle axis. Crucially, in a murine invasive aspergillosis model, BER reduced the fungal burden in lungs and improved survival rates. Thus, we demonstrate that berberine suppresses A. fumigatus growth by disrupting mitochondrial dynamics, elevating reactive ROS, and activating the Hog1-MAPK signaling cascade, ultimately inducing cell cycle arrest. Our findings unveil a previously unrecognized mechanism linking mitochondrial morphology dysregulation to cell cycle control in fungi and establish BER as a promising therapeutic agent targeting mitochondrial-ROS-Hog1 signaling in A. fumigatus infections.

Candida auris is a rapidly spreading multidrug-resistant fungus that causes fatal infections under certain global conditions. Sodium houttuyfonate (SH) and sodium new houttuyfonate (SNH) are stable derivatives of houttuynin (decyl aldehyde) extracted from Houttuynia cordata, both possessing antifungal and antibacterial pharmacological activities. However, the inhibitory effects of SH and SNH on C. auris remain unclear. Therefore, this study aims to evaluate the potential activity and possible mechanisms of SH and SNH as antifungal agents against C. auris. First, our results showed that SH and SNH exhibit significantly inhibitory activity against fluconazole-resistant C. auris strains, but do not possess effective fungicidal activity. In addition, transcriptome and RT-qPCR studies revealed that SH and SNH can repress the expression of genes related to adhesion, aggregation, and biofilm formation. Next, we observed that SH and SNH can disrupt the adhesion and aggregation of early-stage C. auris. Furthermore, using the XTT assay, crystal violet staining, and confocal laser scanning microscopy, we found that the biofilm formation ability of C. auris was disrupted by SH and SNH. We also found that SH and SNH can potentially increase chitin content and expose β-1,3-glucan in the cell wall. Finally, infection models using Galleria mellonella larvae and mice with systemic candidiasis demonstrated that SH and SNH significantly inhibited the colonization and pathological damage of C. auris in vivo. Therefore, our presented results suggest that SH and SNH can effectively inhibit the growth, adhesion, aggregation, and biofilm formation to treat its colonization and pathological damage to the host of C. auris. IMPORTANCE:Recently, the annual proportion of non-C. albicans infections has been rising. The most notable characteristic of C. auris is its resistance to drugs, including multidrug resistance, which results in treatment failures and poses significant challenges in controlling its spread. Sodium houttuyfonate (SH) and sodium new houttuyfonate (SNH) are effective and stable derivatives of houttuynin (decyl aldehyde) extracted from traditional Chinese herbal medicine Houttuynia cordata, both possessing antifungal and antibacterial pharmacological activities. However, the inhibitory effects of SH and SNH on C. auris remain unclear. Through in vitro and in vivo approaches, we have demonstrated that SH and SNH can effectively inhibit the growth, adhesion, aggregation, and biofilm formation to treat its colonization and pathological damage to the host of C. auris. Thus, our findings provide new insights into possible options for clinical applications in the anti-C. auris.

The mitochondrial anchoring protein Num1 directly affects mitochondrial redox function, cell division, and growth in unicellular fungi. However, the functional characterization of Num11, its Candida albicans homolog, remains elusive. Our investigation revealed that Num11 deletion in C. albicans caused profound cellular defects: (1) Disrupted cell cycle progression and mitochondrial dysfunction manifesting as mitochondrial morphological aggregation, ATP depletion, membrane potential collapse, and ROS overproduction; (2) Hypersensitivity to cell wall-perturbing agents accompanied by thicker cell walls and increased surface exposure of β-glucan/chitin; (3) Enhanced macrophage phagocytosis and proinflammatory cytokine release. These cellular alterations translated to significantly attenuated virulence in both Galleria mellonella and systematic mice infection models. Mechanistically, transcriptome profiling and protein interaction analyses demonstrated Num11 deficiency hyperactivates the Cdc42-Cek1 MAPK cascade (phospho-Cek1 increased), driving cell wall remodeling. Our findings establish Num11's dual closely connected regulatory roles in C. albicans pathogenesis: as a mitochondrial scaffold maintaining bioenergetic homeostasis to attenuate growth and as a negative regulator of the Cdc42-Cek1 axis controlling cell wall architecture through affection on mitochondria. These coordinated actions collectively underscore Num11's critical role in mediating host-pathogen interactions during invasive candidiasis.

Ethnopharmacological relevance Vulvovaginal candidiasis (VVC) is a relatively common fungal infectious disease in the female reproductive tract. The pathogenesis of VVC not only involves Candida albicans (C. albicans) infection, but also the improper immune response of the vaginal mucosal immune system to the fungus. As a classical formula, Pulsatilla decoction has been proven to exert protective effect in both clinical and experimental research in VVC. However, the specific mechanism of Pulsatilla decoction in VVC remains elusive. This study investigated the mechanism of n-butanol extract of Pulsatilla decoction (BEPD) in treating VVC from the perspective of type I interferon signaling and related mitochondrial function. Materials and methods A VVC-rat model was developed using an estrogen-based method to evaluate the effectiveness of BEPD in treating VVC, and the therapeutic efficacy of BEPD against VVC was comprehensively evaluated by fungal morphology and burden, neutrophil numbers, histopathology, pro-inflammatory and anti-inflammatory cytokines production, and LDH level. Immunohistochemistry (IHC), immune fluorescence (IF), Western Blot (WB) and RT-qPCR assays were conducted to assess type I interferon signaling and mitochondria functions. Candidalysin-induced vaginal epithelial inflammation in vitro, as cellular models, was employed to detect the changes in type I interferon signaling and mitochondria function before and after BEPD-containing serum intervention. Results BEPD could significantly improve the inflammation, reduce fungal loads and inhibit fungal growth, balance pro-inflammatory and anti-inflammatory cytokine levels in VVC model rats. The findings from IHC, IF, WB and RT-qPCR revealed that BEPD could promote type I interferon signaling and alleviate mitochondrial functional damage in VVC model rats, and BEPD-containing serum could play the same role in vitro. Conclusion The study findings generally demonstrated that BEPD could improve the inflammation and correct the immune imbalance in VVC through regulation of type I interferon signaling and mitochondrial function.

ETHNOPHARMACOLOGICAL RELEVANCE:The combination of Sophorae Flavescentis Radix (Ku Shen) and Cnidii Fructus (She Chuang Zi) has been widely documented in traditional Chinese medicine (TCM) formulations, demonstrating notable antibacterial, anti-inflammatory, and immunomodulatory properties. AIM OF THE STUDY:This study aims to investigate the therapeutic efficacy and mechanistic of the extract of Sophorae Flavescentis Radix-Cnidii Fructus couplet medicines (ESCC) against vulvovaginal candidiasis (VVC) induced by Candida albicans. MATERIALS AND METHODS:A murine model of VVC was established to evaluate vaginal fungal burden and inflammatory cytokine levels, while autophagy-related protein expression and RNA transcriptional profiles were systematically analyzed. A431 cells were used to assess autophagy responses after infection with C. albicans SC5314. Immunofluorescence, quantitative real-time PCR (qRT-PCR), and Western blotting (WB) were utilized to quantify autophagy-related protein expression and transcriptional activity in both murine tissues and A431 cells. Network pharmacology analysis was conducted to predict potential therapeutic targets, followed by in vitro validation of key protein expression levels in the PI3K/AKT/mTOR signaling pathway. RESULTS:In vivo, 120 mg/kg ESCC treatment restored vulvar symptoms to physiological levels in murine models, with the ESCC group exhibiting a significant reduction in vaginal fungal burden. ESCC upregulated LC3B, ATG5, and p62/SQSTM1 expression in vivo and in vitro, reversing the autophagy inhibition caused by C. albicans. Network pharmacology and KEGG enrichment analyses identified the PI3K/AKT/mTOR pathway as a pivotal mechanism, which was further verified by transcriptional and protein-level assays showing that ESCC promoted autophagic flux through pathway inhibition. CONCLUSION:ESCC alleviates C. albicans-induced VVC by activating PI3K/AKT/mTOR-mediated autophagy in vaginal epithelial cells, providing a mechanistic basis for its antifungal and supporting its potential as a plant-derived therapeutic.

The emergence of the "super fungus" Candida auris poses a significant threat to human health, given its multidrug resistance and high mortality rates. Therefore, developing a new antifungal strategy is necessary. Our previous research showed that Baicalein (BE), a key bioactive compound from the dried root of the perennial herb Scutellaria baicalensis Georgi, has strong fungistatic properties against C. auris. Nevertheless, the antifungal activity of BE against C. auris and its mechanism of action requires further investigation. In this study, we explored how BE affects this fungus using various techniques, including scanning electron microscopy (SEM), Annexin V-FITC apoptosis detection, CaspACE FITC-VAD-FMK In Situ Marker, reactive oxygen species (ROS) assay, singlet oxygen sensor green (SOSG) fluorescent probe, enhanced mitochondrial membrane potential (MMP) assay with JC-1, DAPI staining, TUNEL assay and reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Our findings revealed that BE induced several apoptotic features, including phosphatidylserine (PS) externalization, metacaspase activation, nuclear condensation and DNA fragmentation. BE also increased intracellular ROS levels and altered mitochondrial functions. Additionally, transcriptomic analysis and RT-qPCR validation indicated that BE may induce apoptosis in C. auris by affecting ribosome-related pathways, suggesting that ribosomes could be new targets for antifungal agents, in addition to cell walls, membranes, and DNA. This study emphasizes the antifungal activity and mechanism of BE against C. auris, offering a promising treatment strategy for C. auris infection.