Peroxisome proliferator-activated receptor γ (PPARγ) plays a crucial role in the differentiation and maturation of preadipocytes. PPARγ promotes adipogenesis by inducing the expression of fatty acid-binding protein 4 (FABP4). Uncoupling protein 1 (UCP1) is involved in non-shivering thermogenesis and adipocyte browning. The present study aimed to examine the effects of luteolin and apigenin on the gene expression levels and protein concentrations of PPARγ and FABP4, which are involved in adipogenesis, and their effect on UCP1, a thermogenic protein, in the 3T3-L1 preadipocyte cell line. Luteolin and apigenin were prepared at concentrations of 10, 20, and 40 µM and applied to 3T3-L1 preadipocytes during differentiation and maturation. Gene expression levels were measured by real-time PCR, and protein concentrations were measured by ELISA. It was found that the doses used did not cause cytotoxicity in the cells. Luteolin treatment during differentiation and maturation resulted in a decrease in PPARγ and FABP4 gene expression, although the protein concentrations remained unchanged. Additionally, while luteolin treatment did not significantly alter UCP1 gene expression or protein levels during differentiation, it led to a decrease in UCP1 protein concentration during maturation. Apigenin treatment also tended to decrease PPARγ and FABP4 gene expression compared to the control, although no statistical difference was observed. These results suggest that luteolin and apigenin may have regulatory effects on adipogenesis by modulating PPARγ, FABP4, and UCP1 gene expression.

Triple Negative Breast Cancers (TNBCs) are heterogeneous and aggressive tumors with a median overall survival of less than two years. Despite the availability of new drugs, the prognosis remains poor, implicating a more aggressive clinical course in the metastatic setting. This study investigated the effects of metronomic treatment (mCHT) with 5-fluorouracil (5-FU) plus vinorelbine (VNR) on spheroids derived from two different TNBC cell lines (BT-549 and MDA-MB-231) and a patient-derived primary cell line (MS-186). mCHT significantly reduced spheroid growth and altered spheroid architecture, with a pronounced effect in second-generation spheroids, enriched in self-renewing cancer stem cells (CSCs). Expression of CSC-related markers (CD44, CD133, NOTCH-1, and MYC) was more significantly altered-both at the mRNA and protein levels-by mCHT than by standard treatment (STD). In MS-186-derived spheroids, mCHT downregulated EZH2 and STAT3, key regulators of CSC maintenance, and reduced H3K27ac, suggesting a global epigenetic reprogramming. Unlike STD, which partially and transiently reduced stemness markers, mCHT achieved sustained suppression, indicating preferential targeting of therapy-resistant CSCs. These results indicate mCHT as a promising strategy for specifically aiming at the CSC-like compartment in TNBC, underscoring a therapeutic approach that reprograms key epigenetic networks and overcomes resistance to treatment.

Lung cancer (LC) remains the leading cause of cancer-related death in the United States despite advances in therapy. Growth hormone (GH) action has been implicated in tumor progression and therapy resistance across multiple cancers, but its role in LC, particularly non-small cell lung cancer (NSCLC), remains poorly defined. In cancer cells, GH promotes chemoresistance through upregulation of drug-efflux pumps, induction of epithelial-to-mesenchymal transition (EMT), and inhibition of apoptosis. Notably, GH receptor (GHR) expression is significantly elevated in NSCLC compared to normal lung tissue, suggesting a potential therapeutic opportunity. In this study, we investigated the impact of GH action on therapy resistance and tumor progression using integrated transcriptomic analyses and in vitro experiments. Analyses of transcriptomic data from NSCLC patients revealed that high tumoral GHR expression correlates with reduced overall survival, and with upregulation of genes involved in distinct therapy refractory pathways. Our in vitro studies demonstrated that GH promotes chemoresistance in NSCLC cell lines through activation of ABC transporters and EMT pathways, whereas GHR antagonism with the GH receptor antagonist, pegvisomant, effectively counteracts these effects and improves chemotherapy efficacy significantly. Together, our findings identify GHR signaling as a contributor to aggressive and therapy-resistant phenotypes in NSCLC in vitro and suggest that GHR antagonism may enhance chemotherapy sensitivity. These results provide a rationale for further in vivo and mechanistic studies to evaluate the therapeutic potential of targeting GHR in NSCLC.

Aspartame, a widely used artificial sweetener, has been linked to various cancers, including kidney renal papillary cell carcinoma (KIRP). However, the molecular mechanisms underlying this association remain unclear. This study employed network toxicology and molecular docking to investigate potential mechanisms of aspartame-induced KIRP. Differentially expressed genes from TCGA were intersected with aspartame targets and KIRP-related genes, yielding 61 common targets. GO and KEGG analyses revealed enrichment in extracellular matrix degradation, signaling pathways, and immune microenvironment regulation. Univariate Cox regression identified 23 prognostically significant genes, from which multifactorial Cox regression with stepwise selection determined 8 core genes (APLNR, CYP2C19, EDNRA, KLK5, F2R, RAD51, AURKA, and TLR2). A risk model was constructed and validated through VIF analysis, Schoenfeld residual testing, and internal validation using a training-validation split. SHAP analysis identified EDNRA as the primary driver gene. Survival analysis demonstrated that the model effectively stratified KIRP patients, with risk score and tumor stage serving as independent prognostic factors. Molecular docking confirmed stable binding between aspartame and core target proteins. These findings provide mechanistic insights into aspartame-induced KIRP pathogenesis and establish a foundation for future experimental validation.

The Tepetitlán reservoir is affected by untreated domestic wastewater, agricultural runoff, and livestock-related discharges, posing risks to aquatic ecosystems. This study assessed its water quality and evaluated toxic effects on adult zebrafish (Danio rerio). Water samples from six sites (A-F) were analyzed for physicochemical parameters and the presence of metals and pharmaceuticals, quantified at concentrations in the µg L-1 range. While most physicochemical parameters complied with Mexican regulations, true color exceeded established limits, and the presence of contaminants indicated environmental deterioration. Zebrafish were exposed to water from each site for 12, 24, 48, 72, and 96 h. Oxidative stress (OS) biomarkers were measured in the brain, gill, gut, and liver at all time points, and gene expression of antioxidant, detoxification, and apoptosis-related genes was assessed at 96 h in these organs. Significant OS was detected across sites, with increased lipid peroxidation, protein carbonylation, and hydroperoxide levels. Although antioxidant enzymes were activated, their response did not fully counteract oxidative damage. Gene expression analysis revealed upregulation of stress- and apoptosis-related genes. These findings demonstrate that the Tepetitlán reservoir contains pollutant mixtures capable of inducing oxidative and molecular stress responses in zebrafish, underlining potential ecological risks and the need for mitigation efforts.

Background and aims: Low vitamin D3 levels are common in cancer patients, and these patients might benefit from vitamin D3 level normalization in parallel with the conventional oncology treatment. This study aimed to examine the molecular effects of moderate-high-dose vitamin D3 supplementation in vitamin D-deficient cancer patients. Methods: Eight patients under oncological treatment (5 lung cancer, 2 colorectal cancer, and 1 urothelial carcinoma) received 30,000 IU of vitamin D3 per week for two months. Blood samples were collected before and after supplementation, and peripheral blood mononuclear cells (PBMCs) were isolated. With the aim of assessing further potential epigenetic alterations, global DNA methylation level was estimated on the basis of LINE-1 bisulfite-sequencing experiments on cfDNA and PBMC cells. In order to explore the chromatin accessibility alterations after the treatment in PBMCs, an assay for transposase-accessible chromatin with sequencing (ATAC-Seq) was performed using the (10x Genomics, Pleasanton, CA, USA) on a NextSeq 550 instrument using High Output Sequencing kit (Illumina, San Diego, CA, USA). DNA integrity was assessed by the alkaline Comet-assay and telomere qPCR was also performed. Results: After serum 25-hydroxy-vitamin D levels were normalized, DNA integrity in mononuclear cells improved significantly (p = 0.01), while no significant changes were found in granulocytes. Vitamin D3 supplementation also led to significant changes in telomere length in mononuclear cells (p = 0.007). No significant differences were observed in cfDNA levels or DNA methylation in PBMCs and cfDNA after supplementation. ATAC-Seq revealed changes in PBMC composition, including an increased number of NK, pDC cells, and monocytes, especially in patients treated with Pembrolizumab in parallel with vitamin D supplementation. Conclusions: These exploratory findings suggest that the observed immune cell and chromatin changes after vitamin D3 level normalization are compatible with immunomodulatory effects and warrant confirmation in larger, controlled cohorts.

This study compares the ability of the cytokinin (CK) trans-zeatin (tZ) and the CK sugar conjugate 6-(3-methoxybenzylamino)purine-9-arabinoside (BAPA) to induce resistance against the bacterial pathogen Pseudomonas syringae in Arabidopsis thaliana. Treatment with either tZ or BAPA significantly reduced bacterial growth after a later infection. This chemically induced resistance (IR) required the CK receptor AHK3, highlighting its critical role in mediating resistance by tZ and BAPA. This is remarkable as these compounds show either high or no affinity for this CK receptor, respectively. Surprisingly, tZ, but not BAPA, induced the expression of CK response genes, including ARR5, suggesting divergent mechanisms of action. Resistance caused by both compounds was abolished in the npr1 mutant, underpinning the functional relevance of the salicylic acid (SA) signalling pathway. Transcriptomic analysis showed that both BAPA and tZ triggered the expression of distinct sets of genes associated with SA and reactive oxygen species (ROS) but not with jasmonic acid (JA) signalling. BAPA and, to a lesser extent, also tZ activated pattern-triggered immunity (PTI) signalling genes, including genes responsible for PTI signal amplification (PREPIP2) and pathogen-associated molecular pattern (PAMP) signalling (PH1, IDL6). This supported the hypothesis that the PTI pathway mediates the protective effect. Similarities and differences of chemically triggered IR by tZ and BAPA, as well as their potential for application, are discussed.

Fetal development is a critical period that establishes reproductive efficiency and herd performance depending on in-utero epigenetic modifications. Dietary restrictions may affect fetal testis development and the offspring fertility. Several studies have connected genetic instability to circadian cycle disruptions, including epigenetic modifications to melatonin, a key regulator. On day 160 of gestation, 17 male-bearing Brangus heifers were assigned to one of four groups in a 2 × 2 factorial treatment arrangement: adequately fed (ADQ; 100% NRC recommendation, n = 3), nutrient restricted (RES; 60% NRC recommendation, n = 5), or ADQ or RES supplemented with 20 mg/d melatonin (ADQ-MEL, n = 5; RES-MEL, n = 4). On day 240 of gestation, heifers underwent Cesarean sections to collect fetuses and testicular tissues. The fetal testicular tissue was processed and analyzed using the Methyl-MiniSeq Service: Genome-wide bisulfite sequencing (Methyl MiniSeq-GWBS). Sequence reads from Methyl Mini-Seq libraries were identified using standard Illumina platform calling software for methylome profile. RNA-Seq libraries were then sequenced on the Illumina platform for transcriptome profile. The common genes between differentially methylated regions (DMRs) and differentially expressed genes (DEGs) across different treatment groups were identified by an overlap analysis using bedtools v2.31.1. There were 413 DMRs in RES-CON and ADQ-CON testicular tissues, without differential gene expression. Compared with the ADQ-CON group, the ADQ-MEL group showed 411 DMRs and a higher KYAT1 gene expression (P-adj <0.05) without methylation changes. Comparing RES-MEL with RES-CON showed that 9 genes (DAAM1, COL28A1, RPL10, TRPM3, SLIT, ARHGEF40, SYT1, TMEM35B, CSPG4B) were expressed more in the former (P-adj <0.05). The only hypomethylated gene was DAAM1 located on chromosome 10. However, 13 genes (PTPRU, snRNP-E, TMEM59L, MUC5B, ANAPC15, FAM221A, SHCBPiL, PAQR5, PPP4R3C, DTNB, LncRNA, SHANK2, RIC3) showed increased expression in RES-CON vs. RES-MEL without differential methylation alterations, yet there were 370 DMRs. Five genes showed increased expression in RES-MEL compared with ADQ-MEL (P-adj <0.05), including histone H2B on chromosome 23. Two genes (PTPRU, TDRD10) showed increased expression in ADQ-MEL compared with RES-MEL (P-adj <0.05) without affecting methylation and 344 DMRs. In conclusion, dietary melatonin supplementation to nutrient restricted dams may influence fetal development as epigenomic and transcriptomic regulators are altered.

Facioscapulohumeral muscular dystrophy (FSHD) is a progressive and debilitating muscle disease for which no cure currently exists. In the majority of cases, FSHD is associated with the contraction of the D4Z4 macrosatellite repeat array at the 4q35 locus, leading to the inappropriate activation of DUX4, normally expressed during early embryogenesis. In FSHD, the genetic contraction is accompanied by hypomethylation of the D4Z4 array. Although a connection between DNA hypomethylation and DUX4 expression has been suggested, the precise mechanisms that regulate DUX4 transcription remain incompletely defined. The post-translational modification by SUMO was shown previously to repress the expression of Dux, the DUX4 homolog, in mouse embryonic stem cells. Based on these findings, we explored here the contribution of SUMOylation in the regulation of DUX4 in human muscle cells. We demonstrate that TAK‑981 (subasumstat), a selective SUMOylation inhibitor, promotes transcriptional reprogramming of the 4q35 locus and induces DUX4 expression. Importantly, this activation occurs independently of changes in DNA methylation or SMCHD1 ATPase activity. Our findings identify SUMOylation inhibition as a novel regulatory process driving DUX4 expression. This work uncovers the importance of SUMOylation in the epigenetic control of the 4q35 locus and DUX4 transcription, providing a potential therapeutic strategy to modulate DUX4 expression in FSHD.

Concurrent chemotherapy is the standard treatment strategy for advanced-stage nasopharyngeal carcinoma (NPC). However, chemoresistance inevitable develops and the underlying mechanism remains poorly understood. In this study, we identify the arginine methyltransferase PRMT5 as a key gene associated with chemoresistance to paclitaxel in NPC. We demonstrate that PRMT5 facilitated paclitaxel resistance by inducing KCNMB4 expression in nasopharyngeal carcinoma cells. Mechanistically, PRMT5 is recruited to the promoter region of KCNMB4, where it catalyzes H3R2me2s and enhances KCNMB4 expression. Furthermore, elevated levels of PRMT5 or KCNMB4 correlated with poorer survival and higher recurrence rates in NPC patients. Notably, genetic or pharmacological inhibition of PRMT5 significantly sensitized NPC cells to paclitaxel, both in vitro and in vivo. Collectively, these results suggest that the PRMT5-KCNMB4 axis plays a crucial role in mediating chemoresistance in NPC and targeting this axis may provide a promising therapeutic strategy for late-stage NPC patients.

Paediatric acute-onset neuropsychiatric syndrome (PANS) is a syndrome of infection-provoked abrupt-onset obsessive-compulsive disorder (OCD) or eating restriction. Based on the hypothesis that PANS is an epigenetic disorder of immune and brain function, a full-spectrum medicinal cannabinoid-rich low-THC cannabis (NTI164) was selected for its known epigenetic and immunomodulatory properties. This open-label trial of 14 children with chronic-relapsing PANS (mean age 12·1 years; range 4-17; 71 % male) investigated the safety and efficacy of 20 mg/kg/day NTI164 over 12 weeks. Clinical outcomes were assessed using gold standard tools. To define the biological effects of NTI164, blood samples were collected pre- and post-treatment for bulk and single-cell transcriptomics, proteomics, phosphoproteomics, and DNA methylation. NTI164 was well-tolerated, and 12 weeks of treatment decreased the mean Clinical Global Impression-Severity (CGI-S) score from 4·8 to 3·3 (p = 0·002). Significant improvements were observed in emotional regulation (RCADS-P, p < 0·0001), obsessive-compulsive disorder (CYBOCS-II, p = 0·0001), tics (YGTSS, p < 0·0001), attention-deficit hyperactivity disorder (Conner's, p = 0·028), and overall quality of life (EQ-5D-Y, p = 0·011). At baseline, the multi-omic approach revealed that leucocytes from patients with PANS had dysregulated epigenetic (chromatin structure, DNA methylation, histone modifications, transcription factors), ribosomal, mRNA processing, immune, and signalling pathways. These pathways were significantly modulated by NTI164 treatment. NTI164 shows promise as a disease-modifying therapeutic for PANS. Multi-omics reveal broad epigenetic and immune dysregulation in patients, which was modified by NTI164, presenting epigenetic machinery as a therapeutic target in PANS.

Novacetimonas hansenii is a model organism for bacterial cellulose (BC) production, a biopolymer with broad industrial applications. To better understand its carbon source-dependent metabolism, we conducted the first time-resolved transcriptomic analysis of N. hansenii ATCC 23769 grown on five carbon sources (glucose, fructose, mannitol, galactose, and sucrose) across five time points (24-120 h). Glucose, fructose, and mannitol supported growth and efficient BC production, whereas galactose and sucrose resulted in poor yields, likely due to limited transport and enzymatic capacity. Solid-state NMR revealed that cellulose crystallinity and polymorphism were carbon source-dependent, with reduced bcsD expression correlating with lower crystallinity. Transcriptomic profiling showed distinct metabolic responses. Cultures containing glucose and mannitol activated core metabolic pathways, fructose induced a delayed adaptive shift, and galactose and sucrose triggered stress-related gene expression. These results highlight the metabolic plasticity of N. hansenii and offer insights for enhancing BC production through carbon source optomization.

Methyl jasmonate (MeJA) regulates plant development and reproductive processes and significantly influences metabolism. Pears are important economic fruits, but there is still limited research on the changes in primary metabolites in pears after MeJA treatment, and their molecular mechanisms are not yet clear.

The United States is currently facing a drug overdose epidemic. The nucleus accumbens core (NAcore), a brain region critical for reward and aversion behaviors, undergoes structural and functional synaptic adaptations in response to chronic drug exposure. However, the molecular mechanisms underlying these adaptations remain poorly understood. In this study, we investigate the role of dual-specificity phosphatase 5 (DUSP5), a phosphatase known to deactivate extracellular signal-regulated kinase (ERK), in cocaine-induced neuroplasticity. While prior research has linked other DUSP family members to various drugs of abuse, the specific role of DUSP5 in cocaine addiction remains unexplored. We hypothesized that lack of DUSP5 contributes to cocaine-induced maladaptive synaptic plasticity in NAcore. To test this, we employed a rat cocaine self-administration model and molecular analyses and mined publicly available single-cell RNA sequencing data from cocaine-treated NAcore. Our findings reveal a role for DUSP5 in cocaine-related synaptic and behavioral adaptations, highlighting DUSP5 and DUSP5-associated signaling pathways as potential mechanisms underlying substance use disorders and as candidates for therapeutic intervention.

Cisplatin is one of the most effective chemotherapeutic agents used in the treatment of ovarian cancer. However, the frequent development of cisplatin resistance remains a significant limitation, leading to therapeutic failure and poor patient outcomes. Cisplatin cytotoxicity is attributed to the generation of toxic DNA lesions, which can be recognized and processed by a variety of proteins, including the high mobility group box 1 (HMGB1) protein. HMGB1 is a multifunctional protein, which is involved in chromatin remodeling and multiple DNA damage repair pathways. In this study, we investigated the role of HMGB1 in modulating cisplatin sensitivity in human ovarian cancer cells. Using cisplatin-sensitive and cisplatin-resistant human ovarian cancer cell lines, we employed siRNA-mediated HMGB1 knockdown to assess its impact on the cellular responses to cisplatin treatment. In clonogenic survival assays, HMGB1 depletion resulted in a significant reduction in colony formation in cisplatin-resistant cells upon cisplatin exposure, compared with nontargeting siRNA treated cells. Additionally, HMGB1 inhibition significantly enhanced cisplatin-induced apoptosis in the cisplatin-resistant cells. Mechanistically, HMGB1-depleted cells exhibited altered DNA damage responses via modulation of ATM/CHK2 and ATR/CHK1 activity following cisplatin treatment. Notably, DNA immunoblot and modified alkaline comet assay results demonstrated that HMGB1 depletion stimulated cisplatin-DNA adduct formation and impaired the removal of cisplatin-DNA adducts, particularly in the cisplatin-resistant cells. Collectively, these findings uncover novel functions of HMGB1 in mediating cisplatin sensitivity, emphasizing its potential as a therapeutic target to overcome cisplatin resistance in ovarian cancer.

Jasmonic acid (JA) is a critical signal controlling ripening and trait development in non-climacteric (NC) fruit. However, the mechanisms governing the JA biosynthesis remain unclear. Here, the signaling mechanisms for the JA biosynthesis are explored in strawberry (Fragaria vesca), a model NC fruit. The JA biosynthesis is demonstrated to be tightly coupled with the signaling of ABA, a pivotal signal controlling NC fruit ripening. When overexpressed or knocked out by CRISPR/Cas9 editing, FvSnRK2.6, a gene encoding a component of ABA signaling, promotes or inhibits JA production and aroma production, respectively. Moreover, FvSnRK2.6 phosphorylates FvJAZ12, a jasmonate ZIM-domain repressor, at the S142 residue, thereby promoting its degradation. Transforming the FvJAZ12 knockout mutant with FvJAZ12S142A inhibits the production of ABA-induced aroma and JA. Furthermore, our current study reveals that FvMYC2, a transcription factor directly repressed by FvJAZ12, binds to cis-acting elements in the promoters of FvAOC3, FvAOS, FvLOX3, and FvOPR3, thus directly regulating JA biosynthesis. Thus, this study reveals an ABA signaling cascade that leads to JA biosynthesis, thereby elucidating the signaling mechanism governing the JA production during strawberry fruit ripening.

Hypertension is a polygenic, complex disease that impacts men and women differently; whilst the incidence of high blood pressure (BP) is roughly equal over a lifetime, men typically are at higher risk of developing the disease earlier in life, before 50 years of age. There is adequate evidence that the brain is critical for the BP setpoint. The paraventricular nucleus (PVN) of the hypothalamus is an integrative structure that can influence not only neurohumoral responses to blood pressure changes, but also sympathetic drive. Here we manipulate the androgenic status of both male and female spontaneously hypertensive rats (SHRs) to determine how this changes gene expression within the PVN of these animals.

Phosphorus (P) is an essential nutrient for plant growth and development. Phosphate transporter 1 (PHT1) is a transmembrane protein that mediates the uptake and translocation of inorganic phosphate (Pi) in plants. Despite extensive research on the PHT1 family across various species, the PHT1 genes in Eucalyptus grandis are poorly documented and not identified comprehensively.

Nanotechnology and transcriptomics are revolutionizing agriculture by improving sustainability and efficiency. Nanoparticles facilitate the slow release of nutrients for better crop absorption, while transcriptomic analysis uncovers gene expression changes. In a comparative study of zinc-responsive groundnut using bulk ZnSO4 and zinc oxide nanoparticles (nano ZnO), zinc nanoparticles were synthesized from ZnSO4 via a precipitation method. Characterization included particle size analysis (67.5 nm), zeta potential, scanning electron microscopy, energy-dispersive X-ray spectroscopy, and fourier-transform infrared spectroscopy. RNA was isolated from leaf samples treated with either bulk ZnSO4 or nano ZnO after seed treatment and two foliar sprays over 73 days. Transcriptome sequencing with the Oxford Nanopore Sequencer enabled de novo annotation and differential gene expression analysis. In the bulk ZnSO4 versus control comparison, 971 contigs were identified, revealing 792 significant differentially expressed genes (DEGs). For nano ZnO, 971 contigs were also found, with 851 significant DEGs. Pathway network analysis highlighted isoprene biosynthesis as crucial for promoting plant growth and enhancing yield by altering photosynthesis and secondary metabolites. Key genes, DXR and DBR, involved in the isoprene biosynthetic pathway, were identified as vital for isoprene production and zinc metabolism. DBR facilitates redox reactions crucial for producing isoprene precursors and maintaining zinc homeostasis by supporting the function of zinc-containing proteins. Meanwhile, DXR, as a key enzyme in the MEP pathway, links isoprene production to zinc-dependent metabolic processes. Overall, this study suggests that nano-form zinc application enhances the isoprene biosynthetic pathway, leading to improved plant growth and resilience.

Cisplatin is a broad-spectrum anticancer agent. Its main side effect - ototoxicity - may impact the quality of patient's life. Eleutheroside E (EE), the main active component of Acanthopanax, exhibits antioxidant and anti-inflammatory properties. This study investigates the protective effects of EE against cisplatin-induced ototoxicity and its underlying mechanisms. We use C57BL/6 J mice, the House Ear Institute-Organ of Corti 1 (HEI-OC1) cells, and cultured cochlear basement membranes in our experiments. We employ network pharmacology and 4D-FastDIA quantitative proteomic analysis. Our results demonstrate that Cisplatin significantly impairs auditory function in mice. However, EE co-treatment preserves auditory function across most measured frequencies, correlating with reduced damage to cochlear hair cells and spiral ganglion neurons(SGNs). Here, we show that EE attenuates cisplatin-induced pro-inflammatory responses and cellular pyroptosis, possibly via downregulation of the MAPK/NF-κB/NLRP3 signaling pathway. In conclusion, EE may offer a promising strategy for reducing Cisplatin's ototoxicity without affecting its antitumor efficacy.