This year marks the fifth installment of the annual review on bioactivation and reactivity. This series evolved from the broader biotransformation and bioactivation review that started a decade ago, reflecting the increasing importance and interest in this area for drug discovery and development. This year's review features 19 articles that reflect the latest findings and advances in bioactivation and reactivity research. This collection of articles is organized into four general themes: (1) mechanistic insights into CYP-mediated bioactivation, enzyme inhibition, and covalent binding, (2) non-CYP-mediated bioactivation, (3) biological drivers of drug-induced toxicity, and (4) advancements in reactive metabolite trapping.
Animal models are commonly used for prediction of human pharmacokinetics (PKs); however, due to species differences, there are often challenges in translation of the data to clinical outcomes. Humanized mice in the form of chimeric and transgenic models are increasingly proposed as better tools for translational preclinical studies during drug development. Therefore, the aim of this systematic review was to assess the utility and predictive value of humanized mice for human drug metabolism and transport. The databases searched were PubMed, Scopus, Web of Science, and Google Scholar. The inclusion criteria were studies using healthy mice humanized with drug metabolizing enzyme and/or drug transporter and/or nuclear receptors; in original research studies undertaking experimental prediction of human drug transport/metabolism published in the English language. A total of 78 studies were identified and data extracted included experimental groups and number of animals; species and sex; method used to genetically modify the mice; PK data; drug-drug interaction (DDI) predictions and correlations. Discussions and conclusions relied on descriptive summary of results. Chimeric models predominated within the dataset, particularly in PK and metabolite profiling investigations, whereas transgenic mice were more associated with DDI studies. Both model types showed good predictive value for PK and DDIs while chimeric mice replicated human metabolic pathways, identifying human-specific metabolites absent in conventional rodent models. Therefore, this systematic review demonstrates humanized mouse models for drug metabolism and drug transport as useful tools to support prediction of clinical outcomes while recognizing variability in predictive utility across model types and study contexts.
On behalf of my co-authors, I am pleased to present our sixth annual review highlighting articles that have been deemed as impactful in the field of drug transporter sciences. Drug transporters influence the absorption, distribution, and elimination of prescribed drugs and, as a result, it is necessary to understand their impact on new drug development programs. The articles highlighted in this review were selected based on their scientific impact, novelty, and relevance to advancing key areas of transporter research, including mechanistic understanding, translational application, and implications for drug development and regulatory science.Similar to our previous reviews, each co-author has selected peer-reviewed articles and provided a brief synopsis of the article and commentary on the article's significance (Chothe et al. 2021,2022,2023,2024; Sandoval et al. 2025). The review is divided into the following sections: (1) transporter function and DDI evaluation, (2) novel in vivo models, (3) endogenous biomarkers, and (4) drug transporter structure and regulation. The following synopses and commentaries are meant to provide high-level highlights of the articles and readers are encouraged to review these articles for further details. While individual studies are discussed within these categories, a key objective of this review is to place these findings within broader emerging trends in transporter sciences. The synopsis and commentaries are therefore intended not only to highlight individual studies but also to provide perspective on how these advances collectively shape the field.
Tuberculosis (TB), especially in its multidrug-resistant form (MDR-TB), remains a persistent threat to global health despite decades of medical advancements. Standard treatment regimens often involve complex drug combinations, which, while effective, pose a significant risk of drug-drug interactions (DDIs). The growing reliance on complementary and alternative medicine (CAM), particularly herbal supplements, further complicates this landscape by introducing herb-drug interactions (HDIs), many of which are poorly understood or undocumented. These interactions can compromise treatment efficacy, increase toxicity, and hinder patient adherence, particularly in settings where traditional medicine is widely used. This review emphasizes current evidence on the pharmacological interactions between anti-TB/MDR-TB drugs and other conventional medications, as well as commonly used herbal supplements. We explore the pharmacokinetic and pharmacodynamic mechanisms underlying these interactions, with a focus on cytochrome P450-mediated metabolism and modulation of transporter proteins. Highlighted are key interactions involving TB/MDR-TB medication. In addition, the review discusses the emerging role of Network Pharmacology in mapping multi-target interactions, offering new understandings into the potential risks and benefits of integrating herbal medicine into TB care. The potential therapeutic value of select herbs with hepatoprotective and immunomodulatory properties is critically examined alongside safety concerns. By collating and evaluating findings from clinical, pharmacological, and systems-based research, this review aims to guide healthcare professionals in optimizing TB treatment strategies. Emphasis is placed on the need for proactive monitoring, patient education, and culturally sensitive communication regarding herbal use. Integrating evidence-based approaches with traditional practices could enhance treatment outcomes and minimize adverse effects in TB and MDR-TB management.
The present study aims to analyse the curative efficacy of fucoidan on MSG mediated changes in hepatorenal and reproductive organs.For this, 42 adult male rats were grouped into six (N = 7animals/group), and treated for 30 consecutive days with following dose regime: Control (saline water), MSG (60 mg/kg), Fucoidan-low-dose (FLD 100 mg/kg), Fucoidan-high-dose (FHD 200 mg/kg), MSG+FLD (60 mg/kg + 100mg/kg) and MSG+FHD (60 mg/kg + 200mg/kg). On day 31, animals were sacrificed, organs were collected, and subjected to sperm parameters, antioxidant enzyme activities, DNA damage and histomorphometric analysis.The findings showed that MSG caused significant decline in sperm parameters and antioxidant enzymes activity, while increasing the concentrations of oxidative stress markers. MSG administration also showed significant DNA damage in sperm, liver and kidney cells. Additionally, adverse histomorphometric changes including damage to seminiferous tubules, caput and caudal epididymal ducts were noted with decline in sperm concentration. Moreover, hepatorenal histological alterations were identified by leukocytic infiltration, cytoplasmic vacuolation in liver and shrinkage of glomerulus and tubular atrophy in the kidney. On contrary, fucoidan treatment reversed all MSG instigated alterations in dose dependent manner.These findings revealed that fucoidan mitigated MSG induced oxidative, genotoxic and histological alterations in rats, suggesting further mechanistic validation for translational relevance.
Hypertrophic scars (HS) represent a common complication arising from abnormal wound healing following skin injury. Resveratrol and microRNA (miRNA) have been implicated in the formation of HS. However, the precise mechanism by which resveratrol modulates hypertrophic scarring through miRNA regulation remains unclear.Investigating the potential functional effects and molecular mechanisms of resveratrol in HS.RT-qPCR was employed to detect the levels of miR-1290 and ADAMTS8. The targeted binding relationship between miR-1290 and ADAMTS8 was validated through dual luciferase reporter assays and RIP experiments. Cell proliferation, migration capacity, and inflammatory cytokine (IL-1β, IL-6) secretion levels were assessed using CCK-8 assays, Transwell migration assays, and ELISA.MiR-1290 was overexpressed in HS tissue and HSFB cells, whereas ADAMTS8 levels were markedly downregulated, exhibiting a negative correlation between the two. Resveratrol concentration-dependently inhibited miR-1290 levels and upregulated ADAMTS8 levels. ADAMTS8 may represent a direct target gene of miR-1290. Resveratrol may suppress the proliferation, migration, inflammatory cytokine levels, and fibrosis marker levels in HSFB cells. Overexpression of miR-1290 partially reversed the effects of resveratrol, whilst overexpression of ADAMTS8 suppressed the actions induced by miR-1290.Resveratrol may suppress the malignant phenotype of HSFB cells by regulating the miR-1290/ADAMTS8 axis.
HPLC analysis was employed to identify the chemical compositions in BAE. Delphinidin, cyanidin, peonidin, and malvidin were detected in BAE.After establishment of the T2DM mice model, BAE and metformin were treated. Body weight, fasting blood glucose level and glucose tolerance were monitored, followed by detection of β-TG, PF4, CD62p, mitophagy markers. Both BAE and metformin implement reduced the body weight and fasting blood glucose level, and improved glucose tolerance of T2DM mice. BAE implement significantly reduced the β-TG, PF4 and CD62p levels in T2DM mice. FUNDC1, LC3II/LC3I, Pink1, and Parkin protein levels were both significantly elevated in T2DM mice, while Tomm20 expression was remarkedly reduced. BAE implement effectively reversed the FUNDC1, LC3II/LC3I, Pink1, and other relevant markers in T2DM mice.The effect of AMPK pathway involved in the BAE for T2DM treatment was explored. STZ treatment significantly decreased the phospho-AMPK level, while BAE administration reversed the effects of STZ on phospho-AMPK level in mice. Compound C implement effectively abolished the effects of BAE on the expression of platelet activation and mitophagy markers.This study suggests that BAE ameliorates T2DM through inhibiting platelet activation and FUNDC1-mediated platelet mitophagy via the AMPK pathway.
The extensive use of industrial chemicals increases the likelihood of accidental or deliberate environmental release, potentially occurring in occupational, public, or military settings, with skin contact being an important route of exposure. The effectiveness of subsequent skin decontamination depends on the physicochemical properties and absorption profiles of the chemicals involved, as well as the decontamination method applied. This study assessed three solution-based decontamination approaches, hard water, 2% soapy water, and Dahlgren Decon Skin Soap against three representative compounds: chloroacetone, chloroacetonitrile, and crotonaldehyde. For all chemicals tested, statistically significant differences (p < 0.05) were more commonly observed between decontaminated and untreated samples than among the decontamination treatments themselves. These findings indicate that decontamination reliably reduces dermal permeation, even though the specific formulations did not consistently produce distinguishable differences in efficacy. These results support the practical value of prompt decontamination, highlighting that readily accessible solutions can meaningfully mitigate dermal exposure risks in real‑world chemical release scenarios.
Daprodustat is a hypoxia-inducible prolyl hydroxylase inhibitor to treat anaemia in patients with Chronic Kidney Disease (CKD). A stability-indicating RP-HPLC method was developed and validated as per ICH Q2 guidelines. Chromatographic separation was performed on Zorbax SB C18 column (150 mm × 4.6 mm, 3.5 μ) detected at 235 nmusing a PDA detector. A mixture of ethanol and formic acid in a 20:80 (v/v) ratio was used as the mobile phase, with a flow rate of 1 mL/min. Forced degradation studies are conducted according to the International Council of Harmonisation (ICH) guideline Q1A (R2). The HPLC system was coupled to a SCIEX QTRAP 5500 mass spectrometer equipped with an electrospray ionisation interface to identify the plausible structures and fragmentation pattern of all the degradation products.Three degradation products were obtained from forced degradation. Based on the m/z values and molecular formula from the obtained MS spectra, the probable structures of degradation products were elucidated. Plausible degradation mechanism behind the formation of degradation productsThis method can clearly depict the stability of daprodustat under the influence of various environmental factors in less time with low cost. Further, this study also helps in impurity profiling studies.
Semaglutide (SEM) is a GLP-1 analogue, administered subcutaneously or orally. Due to its large molecular structure, it has poor oral absorption and bioavailability. The reported oral bioavailability is 0.4% to 1% in the fasting state. Sodium N-[8-{2-hydroxybenzoyl} amino] caprylate (SNAC) is used as a permeation enhancer to improve gastric permeability and oral bioavailability.This study developed a semi-mechanistic pharmacokinetic (PK) model to predict the steady-state pharmacokinetics of oral SEM, primarily using published clinical data and literature-derived parameters. The present study investigated the impact of SNAC on the gastric absorption of SEM.The semi-mechanistic PK model was developed for the intravenous (IV) and oral formulations. The oral absorption model was developed for SEM at different single doses with varying amounts of SNAC. The dose, SNAC concentration, gastrointestinal permeability, and intestinal first-pass effect impact the PK of the oral SEM. Steady-state PK studies were used to validate the single-dose oral PK model. IV, single-dose, and multiple-dose oral PK models were developed and validated.The developed semi-mechanistic model could be useful for further development of mechanistic, physiologically based pharmacokinetic (PBPK) models for formulation development, drug interactions, and the influence of pharmacokinetics in special populations.
Notoginsenoside R1 (NGR1) shows therapeutic potential in intervertebral disc degeneration (IDD), but its mechanism remains unclear.An in vivo rat tail puncture model and in vitro erastin-induced nucleus pulposus (NP) cells were employed to evaluate the therapeutic effects and underlying mechanisms of NGR1 on IDD. Network pharmacology was performed to predict potential NGR1 targets, which were validated using qRT-PCR. 3-MA (an autophagy inhibitor) and colivelin (a STAT3 activator) were used for mechanistic validation.NGR1 improved disc structure and significantly increased aggrecan and collagen II levels in NP tissues. NGR1 inhibited ferroptosis, as evidenced by increased GPX4 and decreased ACSL4, Fe2+, and ROS levels. NGR1 also promoted autophagy, as indicated by increased Beclin-1 and LC3 II/I levels in NP tissues. Moreover, serum IL-1β and IL-6 were reduced by NGR1 in IDD rats. NGR1 significantly reduced the levels of predicted targets STAT3, IL1B, and MAPK3. In erastin-induced NP cells, NGR1 significantly restored cell viability, enhanced autophagy, and suppressed ferroptosis as well as STAT3, IL1B, and MAPK3 expression, all of which were partially reversed by 3-MA or colivelin.NGR1 alleviates IDD by inhibiting ferroptosis and promoting autophagy, potentially via STAT3 signalling, thereby preserving disc integrity and delaying disease progression.
This study aimed to prepare and evaluate berberine micelles for improving the oral bioavailability and therapeutic effect against skin injury in hyperglycaemic mice. Berberine micelles were fabricated using the film dispersion method with PLGA-PEG-PLGA and glycyrrhizic acid monoamine salt, followed by orthogonal optimisation.The optimised berberine micelles exhibited a particle size of 112.55 ± 2.48 nm, polydispersity index of 0.217 ± 0.005, zeta potential of -31.60 ± 0.23 mV, encapsulation efficiency of 95.57 ± 1.36% (w/w), and drug loading capacity of 7.06 ± 0.25% (w/w). In vitro release of berberine from micelles was higher than that of free berberine across all tested media.Pharmacokinetic studies in rats showed that berberine micelles prolonged the half-life from 5.77 h to 12.19 h and extended the mean retention time from 20.74 h to 143.31 h, with a relative bioavailability 2.64‑fold greater than that of free berberine.In a streptozotocin‑induced hyperglycaemic mouse skin injury model, the high‑dose berberine micelle group significantly accelerated wound closure compared to the free drug group on days 6 and 9, and histopathological analysis of skin, liver, spleen, and pancreas revealed superior tissue recovery. These findings suggest that berberine micelles effectively enhance the anti‑hyperglycaemic and skin‑repairing efficacy of berberine.
Histone hypoacetylation is a well-documented phenomenon associated with nickel (Ni) induced toxic effects, including neurotoxicity. However, the mechanism for histone hypoacetylation in Ni exposure remains ambiguous. Recently, acetyl-CoA availability, especially in the nuclear compartment, for histone acetylation dynamics, is emphasized. In the present study, histone hypoacetylation was evaluated in Ni neurotoxicity cell models, where Ni exposure caused neurite outgrowth impairment. The effects of Ni on histone deacetylases (HDACs) and histone acetyltransferases (HATs), combining with the results of energy metabolites analysis, revealed that the distortion on acetyl-CoA availability was involved in Ni-induced histone hypoacetylation. The function of three nuclear acetyl-CoA synthetases was evaluated, while acetyl-CoA synthetase 2 (ACSS2) nuclear translocation impediment was found in Ni exposure. Promoting ACSS2 nuclear translocation restored Ni-induced H3K9 hypoacetylation and neurites outgrowth impairment. These results indicated that Ni inhibited ACSS2 nuclear translocation to compromise the acetyl-CoA supply for histone acetylation. The impact of Ni on ACSS2 nuclear translocation may provide a new sight into the histone acetylation regulation, suggesting Ni-associated health adverse effects.
The dermal exposure to persistent nerve agents such as VX may lead to skin penetration and uptake in the circulatory system. So far, the efficacy of decontamination lotions to remove VX from the skin has been investigated in vitro using methods for transdermal drug delivery systems such as the vertical diffusion cell, which suffer from several disadvantages. We assessed a full thickness model of human skin placed in inserts of 6-well plates to investigate the efficacy of decontaminants to remove VX. Six different decontamination lotions (reactive skin decontamination lotion (RSDL), the potassium salt of the acetohydroxamic acid (AHAK), sodium hypochlorite (NaOCl) 2% and 10%, soapy water 2% and tap water) were evaluated by calculating the amount of VX that penetrated over 300 min. In addition, the impact of the initiation of decontamination (5 min versus 30 min) was investigated. Early decontamination of VX with AHAK was very effective resulting in a complete decontamination. RSDL and NaOCl 2% showed a substantial decontamination efficacy and the penetrated amount of VX was reduced by 88% (RSDL) and 84% (NaOCl 2%). In contrast, the decontamination efficacy of soapy water and tap water was significantly lower. The delayed decontamination was less effective but still the amount of penetrated VX was significantly reduced. In conclusion, the results underline that the full thickness model of human skin is a suitable tool for investigating the decontamination efficacy of various decontaminants. In addition, the model benefits from low technical requirements.
Bisphenol A (BPA) is a recognised endocrine disruptor resulting in regulatory restrictions and a search for safer alternatives. Bisphenol AP (BPAP), introduced as a BPA substitute, has shown endocrine-disrupting potential and is linked to genotoxicity and recurrent miscarriages. Toxicokinetic evaluation is essential for health risk assessment, yet data on BPAP are lacking. This study characterizes the in vitro and in vivo TK profile of BPAP to facilitate its comparative risk assessment with BPA. A sensitive LC-MS/MS method was developed and validated for quantifying BPAP in rat plasma. In vitro assessments revealed high plasma protein binding in rat and human plasma and indicated metabolism through glucuronidation. Predicted hepatic clearance using the well-stirred model suggested BPAP as a high and intermediate extraction chemical in rats and humans, respectively. In vivo studies showed rapid oral absorption and high clearance (4.24 L/h/kg) after intravenous dosing. BPAP exhibited 18% oral bioavailability. BPAP excretion studies showed feces as major elimination route, with negligible urinary excretion. These findings provide insights into BPAP's disposition profile and highlight that BPAP may not be a safer replacement analogue. This study emphasize the need for further toxicity studies and risk evaluation before considering BPAP a safer BPA alternative.
Epithelial-mesenchymal transition (EMT) is characterized by enhanced fibroblast activation and excessive extracellular matrix deposition, ultimately leading to fibrosis. However, its specific role in silicosis and the underlying mechanisms remain poorly understood. In this study, we investigated the EMT process using a murine model of silicosis and epithelial cell lines (MLE12, A549, and BEAS-2B) exposed to SiO2. Notably, HECTD1 (HECT domain E3 ubiquitin protein ligase 1) was found to be significantly upregulated in epithelial cells during silicosis progression. HECTD1 induced EMT by upregulating mesenchymal markers (α-SMA, Vimentin, and Collagen I) and downregulating the epithelial marker E-cadherin, thereby promoting cell migration and proliferation and ultimately contributing to pulmonary fibrosis. Furthermore, HECTD1 facilitated fibroblast activation through promoting the release of inflammatory cytokines from epithelial cells, thereby exacerbating silicosis. Collectively, our findings establish a link between HECTD1-induced EMT and pulmonary fibrosis, providing new insights into HECTD1 as a potential target for the development of novel therapeutic strategies for silicosis.
Metal release after implantation of medical metal implants can induce serious symptoms of hypersensitivity. A variety of immunologic reactions to metal particles has been demonstrated by a number of studies. To analyze the effect of metal abrasion in the thoracic cavity we aimed at developing a suitable mouse model. However, during the course of the study, high baseline concentrations of both chromium and nickel in conventional husbandry systems were observed. This finding necessitated modifications to the husbandry conditions. Groups of male C57BL/6 J mice were transferred into metal-free cages and fed a diet with reduced nickel and chromium content. After eight weeks, urinary concentrations of chromium and nickel were reduced, which was statistically significant in most groups for chromium and in one group for nickel. Due to their immunological impacts, we report the need to understand potential metal contaminations under conventional husbandry conditions in experimental set ups.
Iron doping has been reported to significantly modify the properties of metal oxide nanoparticles (NPs), including their interaction with cells and therefore toxicity. The present study explores the ability of Fe³ ⁺-doped CeO2 NPs with the varying content of Fe3+ ions (3, 5, and 10 at%) to stimulate eryptosis, a controllable cell death pathway of mature erythrocytes, as an attempt to shed light on hemocompatibility of iron-doped CeO2 NPs. Overall erythrotoxicity of iron-doped CeO2 NPs was evaluated by investigating their ability to trigger spontaneous hemolysis and affect osmotic fragility of rat erythrocytes. Eryptosis of erythrocytes exposed to iron-doped and non-doped CeO2 NPs for 24 h was evaluated by the state-of-the-art flow cytometry-based annexin V staining. Mechanisms involved in iron-doped CeO2 NP-induced eryptosis were evaluated by 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA) and caspase-3 assays, as well as the fluorescent O1O (2-(2'-hydroxy-phenyl)-5-phenyl-1,3-oxazole), NR12S, Fluo-3 AM, BODIPY™ 581/591 C11, and BioTracker Far-red Fe2+ Live Cell imaging probes. Dose-dependent effects of iron-doped CeO2 NPs on hemolysis, osmotic fragility, and eryptosis were revealed. Eryptosis triggered by iron-doped CeO2 NPs was found to be oxidative stress-mediated, cation channel-driven, and caspase-dependent. Oxidative stress and alterations of lipid membranes demonstrated by the tested NPs could be attributable to their direct •OH generation and peroxidase-like activity, as well as Fe2+-mediated Fenton reaction at the surface of Fe³⁺-doped CeO₂ NPs (presumably due to Fe³⁺/Fe²⁺ redox cycling). Importantly, non-doped CeO2 NPs did not promote eryptosis. Nor they stimulated generation of ROS and Ca2+ influx. The presence of iron was found to mediate phosphatidylserine externalization, caspase activation, and changes in lipid membranes of erythrocytes. Notably, Fe³⁺-doped CeO2 NP-induced eryptosis was independent of p38 MAPK and CK1α. Iron-doped CeO2 NPs trigger eryptosis, an effect mediated by iron. Iron doping is a promising modification of CeO2 NPs, which can modulate their toxicity and widen their pharmaceutical profile.
Domestic cats (Felis catus) are among the most popular pets in the world, with the global domestic cat population generally estimated to exceed 600 million and potentially approach 1 billion when feral populations are included. As hypercarnivores, cats exhibit unique metabolic deficiencies, particularly in phase II conjugation enzymes (e.g., glucuronidation, glycine conjugation), which impair elimination of phenolic xenobiotics including pharmaceuticals, feed additives, and contaminants. Consequently, the European Food Safety Authority (EFSA) Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) recommends an additional default uncertainty factor (UF) of 5 for such compounds. Physiologically based kinetic (PBK) modelling offers a mechanistic approach to refine these default factors using chemical-specific kinetic data and such models for the domestic cat are not currently available to the scientific and risk assessment community. Hence, this manuscript focuses on the development and validation of a generic PBK model for the species Felis catus according to the six-step process from the template of the Organisation for Economic Cooperation and Development (OECD) guidance document on characterisation, validation and reporting of PBK models for regulatory purposes. The model integrates meta-analysed physiological parameters from the peer-reviewed literature and 11 perfusion limited compartments. The model has been validated using chemical-specific inputs for 15 pharmaceuticals using in vitro and in vivo clearances to compare in vivo to in vivo and in vitro to in vivo predictions with the available experimental data for plasma maximum concentration (Cmax) and area-under-the-curve (AUC) values in blood after oral and intravenous exposure. Impact of bioavailability on model performance has also been assessed using conservative default values and reported or estimated values. In addition, global sensitivity analysis using the Sobol method identified the muscle:blood partition coefficient as the dominant parameter influencing model output variance. Overall, the generic PBK cat model performed well and most predictions accounting for bioavailability using in vitro derived clearance yielded 86% of Cmax predictions and 64% of AUC predictions were within 2-3-fold of the experimental data as recommended by the OECD. Future applications and refinements of the model with regard to NGRA of food and feed chemicals are highlighted.
Astragaloside IV (AS-IV) represented a promising therapeutic candidate for acute ischaemic stroke. However, its definitive molecular targets and mode of action await full elucidation.The study investigated the protective efficacy of AS-IV and delineated its underlying mechanism in regulating human brain microvascular endothelial cells (BMECs) following oxygen-glucose deprivation/reoxygenation (OGD/R) conditions.Human BMECs were exposed to OGD conditions for 4 h and AS-IV (0-100 µM) was added to the medium immediately for reoxygenating treatment. MiR-320b levels were manipulated 24 h after OGD/R by transient transfection with its inhibitor or mimic. Cell viability and apoptosis were assessed by corresponding kits. The expression level of miR-320b or PTEN was achieved by RT-qPCR. Pro-inflammatory cytokines were detected by ELISA.AS-IV rescued cells under OGD/R condition by enhancing cell viability, curbing apoptosis and pro-inflammatory signalling. This aligned with the pro-survival signature of miR-320b upregulation in OGD/R cells. Conversely, miR-320b knockdown abrogated the AS-IV efficacy. RIP and dual-luciferase reporter assay confirmed PTEN as the direct downstream target of miR-320b.AS-IV protected cells under OGD/R conditions by improving viability and inhibiting inflammation as well as apoptosis via miR-320b/PTEN axis.
