Experimental Model Research Articles

Proteomic analysis of the sponge Aggregation Factor implicates an ancient toolkit for allorecognition and adhesion in animals

The discovery that sponges (Porifera) can fully regenerate from aggregates of dissociated cells launched them as one of the earliest experimental models to study the evolution of cell adhesion and allorecognition in animals. This process depends on an extracellular glycoprotein complex called the Aggregation Factor (AF), which is composed of proteins thought to be unique to sponges. We used quantitative proteomics to identify additional AF components and interacting proteins in the classical model, Clathria prolifera, and compared them to proteins involved in cell interactions in Bilateria. Our results confirm MAFp3/p4 proteins as the primary components of the AF but implicate related proteins with calx-beta and wreath domains as additional components. Using AlphaFold, we unveiled close structural similarities of AF components to protein domains in other animals, previously masked by the mutational decay of sequence similarity. The wreath domain, believed to be unique to the AF, was predicted to contain a central beta-sandwich of the same organization as the vWFD domain (also found in extracellular, gel-forming glycoproteins in other animals). Additionally, many copurified proteins share a conserved C-terminus, containing divergent immunoglobulin (Ig) and Fn3 domains predicted to serve as an AF-interaction interface. One of these proteins, MAF-associated protein 1, resembles Ig superfamily cell adhesion molecules and we hypothesize that it may function to link the AF to the surface of cells. Our results highlight the existence of an ancient toolkit of conserved protein domains regulating cell-cell and cell-extracellular matrix protein interactions in all animals, and likely reflect a common origin of cell adhesion and allorecognition.

A new protocol for the induction of chronic mastitis with intramammary infusion of lipopolysaccharide (LPS) in Balb/c mice

Mastitis is inflammation of the mammary tissue and is commonly observed in farm animals. The problem causes severe financial losses in the dairy industry in terms of veterinary costs, milk disposal, and treatment expenses. Bacteria are the main actors in the etiology and cause acute and chronic inflammatory changes in the mammary tissue. Acute inflammatory changes are easily recognized clinically, and treatment is initiated immediately, but subacute inflammation progresses insidiously and leads to chronic inflammation with irreversible fibrotic changes. Standardized experimental models for the induction of acute mastitis in laboratory animals are available. Usually, infusion of bacteria or some bacterial structural components into mammary tissue is easily applied for this purpose. However, there are few studies on the induction of chronic mastitis with fibrotic changes, and the applications are relatively complex. In this study, LPS was infused through the teat duct three times on days 0, 5, and 10 to induce chronic mastitis in mice. Tissues were sampled on days 1, 6, and 15 to evaluate histopathological changes. While severe neutrophil infiltrates, a component of acute inflammation, were observed on day 1, lymphocyte infiltrates increased on day 6, consistent with subacute inflammation. On day 15, lesions representing chronic mastitis, such as fibrosis and lymphocyte infiltration, were observed. A model similar to the lesions in chronic mastitis of dairy cattle was successfully and easily established by LPS infusion in mice.

Involvement of autophagy in germ cells in an experimental model of varicocele in rats before and after varicocelectomy.

Autophagy, a catabolic process enabling cellular organelles and proteins' reuse for energy, has been observed in varicocele models, but the effect of surgical treatment on this process remains unknown. This study aims to assess autophagy in varicocele models undergoing surgical correction. Twenty-one adolescent male rats were induced with varicocele and divided into three groups: sham, varicocele, and varicocele with varicocelectomy. After 21days, testicles were examined histologically for spermatogenesis (Jonhsen's score) and immunohistochemically for autophagy markers (LC3A, Beclin-1, Ambra-1, ULK-1, p62). Positive germ cells were quantitatively evaluated, and data were statistically analyzed (p < 0.05). Histological examination revealed significantly reduced Jonhsen's scores in varicocele compared to sham and varicocelectomy groups (p < 0.05). Expression of autophagy markers (LC3A, Beclin-1, Ambra-1, ULK-1, p62) was significantly higher in varicocele than sham and varicocelectomy groups (p < 0.05), and in varicocelectomy than sham (p < 0.05). Varicocele activates autophagy markers, with p62 potentially modulating autophagy despite being considered an inhibitor. While varicocelectomy improves histology, it doesn't fully inhibit autophagy, suggesting ongoing germ cell dysfunction despite treatment. This underscores varicocele's detrimental effects on germ cell functionality.

Isoproterenol mechanisms in inducing myocardial fibrosis and its application as an experimental model for the evaluation of therapeutic potential of phytochemicals and pharmaceuticals.

Cardiac injury initiates repair mechanisms and results in cardiac remodeling and fibrosis, which appears to be a leading cause of cardiovascular diseases. Cardiac fibrosis is characterized by the accumulation of extracellular matrix proteins, mainly collagen in the cardiac interstitium. Many experimental studies have demonstrated that fibrotic injury in the heart is reversible; therefore, it is vital to understand different molecular mechanisms that are involved in the initiation, progression, and resolution of cardiac fibrosis to enable the development of antifibrotic agents. Of the many experimental models, one of the recent models that has gained renewed interest is isoproterenol (ISP)-induced cardiac fibrosis. ISP is a synthetic catecholamine, sympathomimetic, and nonselective β-adrenergic receptor agonist. The overstimulated and sustained activation of β-adrenergic receptors has been reported to induce biochemical and physiological alterations and ultimately result in cardiac remodeling. ISP has been used for decades to induce acute myocardial infarction. However, the use of low doses and chronic administration of ISP have been shown to induce cardiac fibrosis; this practice has increased in recent years. Intraperitoneal or subcutaneous ISP has been widely used in preclinical studies to induce cardiac remodeling manifested by fibrosis and hypertrophy. The induced oxidative stress with subsequent perturbations in cellular signaling cascades through triggering the release of free radicals is considered the initiating mechanism of myocardial fibrosis. ISP is consistently used to induce fibrosis in laboratory animals and in cardiomyocytes isolated from animals. In recent years, numerous phytochemicals and synthetic molecules have been evaluated in ISP-induced cardiac fibrosis. The present review exclusively provides a comprehensive summary of the pathological biochemical, histological, and molecular mechanisms of ISP in inducing cardiac fibrosis and hypertrophy. It also summarizes the application of this experimental model in the therapeutic evaluation of natural as well as synthetic compounds to demonstrate their potential in mitigating myocardial fibrosis and hypertrophy.

Effectiveness of Modified Truth or Dare Game in Improving Toxicology Knowledge of Workers at Tofu Factory X

There are 84,000 tofu producers in Indonesia, ranging from household industries with 5-8 workers to medium-sized industries with 50-100 workers. In a single tofu production process, additional chemicals such as CH3COOH and MgSO4 are used. These conditions pose potential safety and health hazards for workers, requiring control measures. This study employs a quantitative approach, using an experimental model with treatment in the form of a card game. Data was collected using primary data methods, such as questionnaire instruments and direct field observation. The questionnaire was compiled based on priority hazard assessment using HIRADC calculations and then tested for validity and reliability. The questionnaire results were tested using the Wilcoxon Signed Rank Test because the data were not normally distributed. The study results for worker respondents (0.027) and the public (0.000), or P value &lt; 0.05, indicate that H0 is rejected, and Ha is accepted. The modified truth-or-dare card game method was found to have a significant impact on toxicology knowledge among workers and the general public.

Deepening the Role of Pectin in the Tissue Assembly Process During Tomato Grafting.

Cell walls play essential roles in cell recognition, tissue adhesion, and wound response. In particular, pectins as cell-adhesive agents are expected to play a key role in the early stages of grafting. To test this premise, this study focused on examining the dynamics of the accumulation and degree of methyl-esterification of pectic polysaccharides at the graft junctions using tomato autografts as an experimental model. Monosaccharide analysis showed a marked increase in homogalacturonan from 25% to 32 or 34% at the junction zones early after grafting. In addition, a decrease in the degree of homogalacturonan methyl-esterification up to 38% in the scion and 64% in the rootstock was observed in the first few days after grafting, accompanied by an increase in pectin methyl-esterase activity of up to 20-30% in the tissues surrounding the graft junction. These results shed light on the role of homogalacturonan in grafting and reinforce the key function of pectin as one of the most relevant cell wall components during the grafting process.

Colchicine Alleviates Interstitial Lung Disease in an Experimental Autoimmune Myositis Murine Model by Inhibiting the Formation of Neutrophil Extracellular Traps.

Our previous study has shown that neutrophil extracellular traps (NETs) were associated with idiopathic inflammatory myopathy-related interstitial lung disease (IIM-ILD). Colchicine plays an anti-inflammatory role mainly by inhibiting the activity and chemotaxis of neutrophils. This study aims to verify therapeutic effects and mechanism of colchicine in IIM-ILD. 20 experimental autoimmune myositis (EAM) model mice were randomly divided into EAM group, colchicine groups (1, 2mg/kg) and Cl-amidine group (positive control), five mice in the control group received sham modeling procedure. After 5weeks, the mice were sacrificed to evaluate the degree of pulmonary interstitial lesions and the formation of NETs. Human neutrophils were pretreated with colchicine (40nmol/L) and stimulated to form NETs. Human pulmonary microvascular endothelial cells (HPMECs) were pretreated with colchicine and stimulated with NETs, and markers of inflammation and pyroptosis were detected. Pathological staining of lung tissue showed that severity of ILD and NETs infiltration were significantly alleviated in colchicine groups (P < 0.01) and in the Cl-amidine group (P < 0.01), and the serum level of NETs was also significantly decreased in colchicine groups (P < 0.05) and in the Cl-amidine group (P < 0.05). Colchicine intervention significantly attenuated PMA-induced NETs formation in vitro (P < 0.0001). Colchicine intervention significantly reduced marker expressions of inflammasome and pyroptosis in HPMECs stimulated by NETs and in the lung tissue of EAM mice. Colchicine can alleviate ILD in EAM mice by inhibiting NETs formation, inflammasome activation and endothelial cell pyroptosis. These findings provide a basis for targeting NETs and colchicine administration in the treatment of myositis-associated ILD.

PET Granule Replacement for Fine Aggregate in Concrete and FRP-Wrapping Effect: Overview of Experimental Data and Model Development

In this study, polyethylene terephthalate (PET) was substituted for 10%, 20%, and 30% of the sand volume in concrete. Compressive, splitting tensile, and flexural strength tests were applied to the concrete samples and stress–strain graphs were obtained. It was observed that PET substitution caused a decrease in the mechanical properties of the concrete. For this reason, the concrete with the best PET substitution rate (10%) was reinforced by wrapping it with carbon fiber-reinforced polymer (CFRP) and glass fiber-reinforced polymer (GFRP), and the same experiments were repeated. It was observed that a 10% PET substitution reduced the strength of the reference concrete by about 6%. However, wrapping the PET-substituted concrete with CFRP and GFRP increased the strength by about 1.9 and 1.5 times, respectively, surpassing that of the reference sample. In addition, this study provides a comprehensive database by bringing together experimental data from studies in which PET was used as a substitute by volume or weight instead of fine aggregate in concrete. The models proposed in this study, along with previous models, were tested for applicability. Similarly, the model suggestions in the literature for fiber-reinforced polymer (FRP)-confined concrete were tested with the experimental data in this study, and their suitability for PET-substituted concrete was discussed.

Unveiling the Role of Resident Memory T Cells in Psoriasis.

Psoriasis is a chronic inflammatory skin disease characterized by periods of remission and relapse. In this pathology, keratinocytes, dendritic cells, and different subpopulations of T cells are critical to developing psoriatic lesions. Although current treatments can reduce symptoms, they reappear in previously injured areas months after stopping treatment. Evidence has pointed out that besides T helper 17 cells, other T cell subsets may be involved in relapses. This review focuses on the leading evidence linking resident memory T cells and P2X7 receptor to psoriasis' pathogenesis and their role in this pathology. Finally, we discuss some of the most widely used experimental murine models and novel strategies to investigate further the role of resident memory T cells in psoriasis.

Nonlinear spectral dispersion in resonant Auger scattering from SF6 for studying nuclear potentials and dynamics

In this study, we integrate experimental observations and theoretical models to elucidate the complex phenomena observed in the resonant S K-edge Auger scattering spectra of the SF6 molecule. A two-dimensional spectral map, constructed of incident photon energy and kinetic energy of the emitted Auger electron, is shown to be a versatile tool for understanding a character of the core-excited potential energy surface and change of the molecular geometry. Our findings reveal how the distinct dispersion behavior of multiple spectral lines enables mapping of ultrafast dynamics within the short-lived core-excited states. Our results confirm the presence of nuclear dynamics in the S1s−16a1g1 and S1s−16t1u1 core-excited states, while dynamics is absent in the S1s−17t1u1 state. Using a combination of analysis, simulations with Coulomb model potentials, and a simple analytical approximation, we qualitatively demonstrate how the varying characteristics of spectral dispersion—classified as Raman, non-Raman, and anti-Raman—mirror the relative gradients of the intermediate and final states in the resonant x-ray scattering process. This insight allows for the effective mapping of molecular potential energy curves, offering a prospective tool on the underlying mechanisms of resonant Auger scattering and its potential for probing molecular dynamics. Published by the American Physical Society 2024

Enhanced phase prediction of high-entropy alloys through machine learning and data augmentation.

The phase structure information of high-entropy alloys (HEAs) is critical for their design and application, as different phase configurations are associated with distinct chemical and physical properties. However, the broad range of elements in HEAs presents significant challenges for precise experimental design and rational theoretical modeling and simulation. To address these challenges, machine learning (ML) methods have emerged as powerful tools for phase structure prediction. In this study, we use a dataset of 544 HEA configurations to predict phases, including 248 intermetallic, 131 solid solution, and 165 amorphous phases. To mitigate the limitations imposed by the small dataset size, we employ a Generative Adversarial Network (GAN) to augment the existing data. Our results show a significant improvement in model performance with data augmentation, achieving an average accuracy of 94.77% across ten random seeds. Validation on an independent dataset confirms the model's reliability and real-world applicability, achieving 100% prediction accuracy. We also predict FCC and BCC phases for SS HEAs based on elemental composition, achieving a peak accuracy of 98%. Furthermore, feature importance analysis identifies correlations between compositional features and phase formation tendencies, which are consistent with experimental observations. This work proposes an effective strategy to enhance the accuracy and generalizability of machine learning models in phase structure prediction, thus promoting the accelerated design of HEAs for a wide range of applications.

Chromosomal rearrangements and instability caused by the LINE-1 retrotransposon.

LINE-1 (L1) retrotransposition is widespread in many cancers, especially those with a high burden of chromosomal rearrangements. However, whether and to what degree L1 activity directly impacts genome integrity is unclear. Here, we apply whole-genome sequencing to experimental models of L1 expression to comprehensively define the spectrum of genomic changes caused by L1. We provide definitive evidence that L1 expression frequently and directly causes both local and long-range chromosomal rearrangements, small and large segmental copy-number alterations, and subclonal copy-number heterogeneity due to ongoing chromosomal instability. Mechanistically, all these alterations arise from DNA double-strand breaks (DSBs) generated by L1-encoded ORF2p. The processing of ORF2p-generated DSB ends prior to their ligation can produce diverse rearrangements of the target sequences. Ligation between DSB ends generated at distal loci can generate either stable chromosomes or unstable dicentric, acentric, or ring chromosomes that undergo subsequent evolution through breakage-fusion bridge cycles or DNA fragmentation. Together, these findings suggest L1 is a potent mutagenic force capable of driving genome evolution beyond simple insertions.

Understanding and Using Animal Models of Hepatotoxicity.

Hepatotoxicity is a critical health hazard, primarily contributing to the increased incidence of deaths globally. The liver is one of the major and extremely vital organs of the human body. Autoimmune diseases, viruses, exposure to toxicants such as carcinogens, and changes in eating habits can all cause liver problems, among other things. Free radical generation, together with raised enzyme levels including SGOT, SGPT, and total bilirubin, are among the pathological changes set off by liver injury. Hepatotoxicity causes changes in cells, such as eosinophilic cytoplasm, nuclear pyknosis, fatty degeneration, too many liver lesions, and hepatic centrilobular necrosis due to lipid peroxidation. Researchers have used animal models to investigate liver diseases and toxicities. Drugs such as azathioprine, alcoholism, paracetamol intoxication, and anti-tuberculosis drugs are some of the most common causes of liver toxicity. These toxins cause calcium ions (Ca2+), reactive oxygen species (ROS), and inflammatory mediators to be released inside cells. This activates immune cells like NK cells, NKT cells, and Kupffer cells. These signaling pathways also play roles in hepatotoxicity. Due to its pathogenesis, no effective drug is currently available for hepatotoxicity due to a lack of understanding related to the signaling factors involved in it. The paper primarily examines different experimental models of hepatotoxicity, including non-invasive and invasive methods, as well as genetic models. As such, these models are crucial tools in advancing our understanding of hepatotoxicity, thus paving the way for new therapeutic interventions.

Dietary fiber pectin alters the gut microbiota and diminishes the inflammatory immune responses in an experimental peach allergy mouse model.

Since therapeutic options are limited the utilization of prebiotics is suggested to prevent food allergies (FAs). Using an experimental peach allergy model we explored the effect of dietary fiber pectin, a high-methoxyl heteropolysaccharide, on the manifestation of FA. CBA/J mice were sensitized, subsequently orally boosted and provoked with peach peel extract. For dietary intervention, mice were fed a pectin containing diet before (primary-preventive) or after (secondary-preventive) sensitization. Non-treated allergic and sham-treated mice were fed a diet containing 20% cellulose. Fecal microbiota, humoral and intestinal immune cell responses were analyzed. Pectin remarkably affected the gut microbiota composition and diversity, promoting mainly the growth of Bacteroides. The frequency of mast cells, macrophages, and CD3+T cells in the lamina propria of the small intestine was reduced, whereas the frequency of B cells and CD4+T cell subpopulation was enhanced. Pectin intervention in the primary-preventive stetting significantly triggered serum IgA levels, whereas production of IgE and mMCPT-1 was reduced. Remarkably, in both settings peach allergen-specific IgG1/IgG2a ratio and specific IgE were significantly reduced to baseline. The data suggest, that dietary supplementation of pectin in both intervention approaches can diminish inflammatory responses and signs of allergic immune responses, accompanied by alteration of the gut microbiota composition.

Effect of vitamin K1 supplementationon coronary calcifications in hemodialysispatients: a randomized controlled trial.

Chronic kidney disease (CKD) is associated with several adversecardiovascular outcomes, including coronary heart disease, heart failure, and arrhythmias. The severityofarterial calcifications predicts the risk of coronary heart disease and increases the risk of premature cardiovascular death. In experimental models, vitamin K1 supplementationappears to reduce coronary artery calcifications. In this single-center clinical trial (NCT04247087 on 07/09/2019), we randomized 60 Mexican patients onchronic hemodialysis and a coronary calcification score> 10 Agatston units to receive 10mg intravenous vitamin K1 or placebo at the end of the hemodialysis sessionthrice weekly for 12months. The primary outcome was the progression of coronary artery calcifications as assessed by the absolute change in Agatston and coronary calcium volume scores. The baseline coronary calcium score was112.50 (14-2027) Agatston units in the vitamin K1 group and 177 (10-2843); Agatston units in the placebo group (p = 0.71), and after 12months, the coronary calcium score in the vitamin K1 group was78.50 (10-1915) Agatston units in the vitamin K1 group versus 344 (10-3323); Agatston units (p = 0.05) in the placebo group.Progression of coronary calcification was 20.8% in the vitamin K1 group versus 44% in the placebo group, with a relative risk (RR) of 0.45 (CI 95% 0.18-1.15). In the Mexican hemodialysis cohort enrolled in this studyintravenous vitamin K1 supplementationreduced the progression of coronary artery calcifications by 55% compared with placebo over a 12-month follow-up period.

Experimental and pilot model research of spiral cyclone with curvilinear channels

The advanced type of cyclone was applied to separate wood particulate matter from the air. The cyclone was designed in the laboratory of Vilnius TECH. A series of experimental studies was conducted both in the lab and under industrial conditions. These studies aimed to specify the air velocity and aerodynamic resistance in the experimental and pilot six-channel cyclone with spiral casings and curvilinear semi-rings. Air treatment efficiency was also determined. The highest air treatment efficiency achieved using experimental cyclone was 91.4%, while the pilot cyclone achieved an efficiency of 94. 5%. The industrial and experimental cyclones were found to have similar air treatment efficiency

Chemerin and Polycystic Ovary Syndrome: A Comprehensive Review of Its Role as a Biomarker and Therapeutic Target.

Background: Chemerin, an adipokine implicated in inflammatory, metabolic, and adipogenic processes, has been detected in high serum concentration in women with polycystic ovary syndrome (PCOS) and seems to play a role in PCOS pathogenesis. Moreover, at present, no comprehensive and critical document is available in the literature on this topic. The aim of the current study was to comprehensively review the latest available data to confirm the evidence about the association between chemerin and PCOS, highlighting its potential role as an upcoming biomarker and therapeutic target. Methods: A search in the literature of studies published between 2019 and 2024 was conducted using PubMed, Cochrane Library, and Web of Science, focusing on research related to chemerin, PCOS, and PCOS-related features, comorbidities, and complications. A qualitative structured synthesis of key findings was performed according to the specific thematic areas selected, including and discussing clinical data on women with PCOS and experimental studies in humans and animal models of PCOS. Results: Available data confirm increased serum levels of chemerin in women with PCOS compared with controls, independent of obesity and body mass index. Chemerin is associated with insulin resistance, hyperandrogenism, and ovarian dysfunction in PCOS individuals, inhibiting folliculogenesis and steroidogenesis. Experimental animal models underscore chemerin's regulatory roles through its receptors within the hypothalamic-pituitary-ovarian axis and peripheral tissues. High systemic levels of chemerin in PCOS may also be related to the increased risk of pregnancy complications, especially gestational diabetes mellitus and preeclampsia. Conclusions: The current review study highlights the role of chemerin in PCOS pathophysiology, severity, and associated comorbidities and complications, assessing its value as a future biomarker and foreshadowing its potential as a therapeutic target.

Aged Garlic Extract (AGE) and Its Constituent S-Allyl-Cysteine (SAC) Inhibit the Expression of Pro-Inflammatory Genes Induced in Bronchial Epithelial IB3-1 Cells by Exposure to the SARS-CoV-2 Spike Protein and the BNT162b2 Vaccine.

Garlic (Allium sativum L.) is a species of the onion family (Alliaceae) widely used as a food and a folk medicine. The objective of this study was to determine the effects of AGE (aged garlic extract) on pro-inflammatory genes relevant to COVID-19. To this aim, we treated bronchial epithelial IB3-1 cells with SARS-CoV-2 spike protein (S-protein) or with the COVID-19 BNT162b2 vaccine in the absence or in the presence of AGE. The results obtained demonstrated that AGE is a potent inhibitor of the S-protein-induced expression of the IL-1β, IL-6 and IL-8 genes. Bio-Plex analysis demonstrated that AGE reduced release of IL-6 and IL-8, which were highly induced by S-protein. No inhibition of cells' growth, toxicity and pro-apoptotic effects were found in AGE-treated cells. The effects of one of the major AGE constituents (S-allyl cysteine, SAC) were studied on the same experimental model systems. SAC was able to inhibit the S-protein-induced expression of IL-1β, IL-6 and IL-8 genes and extracellular release of IL-6 and IL-8, confirming that S-allyl-cysteine is one of the constituents of AGE that is responsible for inhibiting S-protein-induced pro-inflammatory genes. Docking experiments suggest that a possible mechanism of action of SAC is an interference with the activity of Toll-like receptors (TLRs), particularly TLR4, thereby inhibiting NF-κB- and NF-κB-regulated genes, such as IL-1β, IL-6 and IL-8 genes. These results suggest that both AGE and SAC deserve further experimental efforts to verify their effects on pro-inflammatory genes in SARS-CoV-2-infected cells.

Inter-organ communication: pathways and targets to cardioprotection and neuro-protection. A report from the 12th Hatter Cardiovascular Institute workshop.

A long-standing aim in the setting of various pathologies including acute myocardial infarction, chronic kidney disease (CKD), and ischaemic stroke, has been to identify successful approaches to augment cellular and organ protection. Although the continual evolution and refinement of ideas over the past few decades has allowed the field to progress, we are yet to realise successful clinical translation of this concept. The 12th Hatter Cardiovascular Workshop identified a number of important points and key questions for future research relating to cardio- and neuro-protection and interorgan communication. Specific topics that were discussed include the 'cardio-metabolic-renal' axis of organ protection, the parasympathetic signalling hypothesis, the role of the coronary microvasculature in myocardial infarction, the RISK pathway of cardioprotection, extracellular vesicles and the way forward, the future for clinical studies of remote ischaemic conditioning, and new experimental models for cardio-oncology investigations.

Repurposing Dapagliflozin for Mitigation of the Kidney Injury Triggered by Cadmium in Rats: Role of Autophagy, Apoptosis, and the SIRT1/Nrf2/HO-1 Pathway.

Background/Objectives: The antioxidant/antiapoptotic features of dapagliflozin (DPG) have mediated its beneficial actions against several experimental models. However, no studies have been conducted to determine whether DPG mitigates the renal injury triggered by cadmium (Cd). Herein, DPG was studied for its potential to attenuate kidney damage in Cd-intoxicated rats, as well as to unravel the mechanisms involving oxidative events, autophagy, and apoptosis. Methods: Histopathological analysis, immunohistochemical staining, and ELISA were conducted on kidney tissue samples. Results: Cd administration (5 mg/kg/day; p.o.) prompted significant renal damage, as evidenced by histopathological changes, elevated kidney injury molecule-1 (KIM-1) expression, and increased serum creatinine and urea. Interestingly, DPG (1 mg/kg/day; p.o.) significantly mitigated these harmful effects without affecting renal Cd metal accumulation. Mechanistically, DPG curbed Cd-induced renal pro-oxidant response and stimulated the antioxidant sirtuin 1 (SIRT1)/nuclear factor (erythroid-derived 2)-like 2 (Nrf2)/heme oxygenase 1 (HO-1) axis. Moreover, DPG restored autophagy by decreasing sequestosome-1/protein 62 (SQSTM-1/p62) accumulation and stimulating the AMP-activated protein kinase (AMPK)/mechanistic target of rapamycin (mTOR) pathway. In tandem, DPG suppressed Cd-induced apoptosis by lowering renal Bcl-2 associated-x protein (Bax) and cytochrome C (Cyt C) levels and caspase 3 activity. Conclusions: These findings indicate that DPG attenuates Cd-induced nephrotoxicity by enhancing the SIRT1/Nrf2/HO-1 antioxidant pathway, promoting AMPK/mTOR-directed autophagy, and inhibiting apoptotic cell death.