This study was carried out to screen PANoptosis-related biomarker genes and the involved underlying mechanism in clear cell renal cell carcinoma (ccRCC). Differentially expressed (DE) PANoptosis-related genes were identified. Then prognostic biomarker genes were screened, and used to construct a prognostic model. The predictive ability of the model was explored by receiver operating characteristic (ROC) curves. Then the correlation between risk score with clinical features and immune cell infiltration were evaluated. The expression level and prognosis of gelsolin (GSN) were examined in datasets. Furthermore, in vivo and in vitro experiments were employed to investigate underlying mechanisms. ScRNA-seq analysis uncovered that PANoptosis scores varied significantly across 9 cell types in the tumor microenvironment. A total of 26 DE-PANoptosis-related genes were identified by intersecting DEGs and PANoptosis-related genes. Then 3 prognosis biomarker genes were screened and used to build the prognostic model, namely TLR3, GSN and TNFRSF1A. The risk score was associated with tumor grade, stage (I, II, III, IV), pathologic T-Stage, etc, and also significantly correlated with immune cells and expression of immune checkpoint proteins. In addition, GSN was upregulated in ccRCC, and high expression of GSN indicated a favorable OS. In vivo and in vitro experiments revealed that GSN liquid-liquid phase separation (LLPS) directly induces PANoptosis in ccRCC by inhibiting YAP-dependent glycolysis. In conclusion, using TLR3, GSN and TNFRSF1A, the established PANoptosis-related prognostic model was established and showed good predictive performance for predicting prognosis of ccRCC patients. GSN LLPS directly induces PANoptosis in ccRCC by inhibiting YAP-dependent glycolysis.

Ischemic diseases represent a broad and complex group of clinical pathologies caused by acute or chronic tissue perfusion disorders [...].

In this study, in a culture of renal epithelial cells, we identified those expressing nestin, a cytoskeletal protein associated with stem/progenitor/activated/proliferating cell states. A mouse expressing GFP under the nestin promoter was used, followed by cell isolation and culture. It is hypothesized that this can be used to assess the stem/progenitor/activated/proliferating cell level in a mixed kidney cell culture. Both nestin-positive and nestin-negative cells were demonstrated to be present in the culture. After visualization, cells were attached to a glass slide with a grid, fixed, and prepared for electron microscopy analysis, with each cell visually identified by light microscopy being analyzed. Electron microscopy revealed tight interactions between nestin-positive and nestin-negative cells. Significant differences in the ultrastructure of nestin-positive and nestin-negative cells were observed. Nestin-positive cells were distinguished by a high ribosome content, indicating high protein-synthesizing activity. In the nestin-GFP-high (sorted) population examined by electron microscopy, vesicle-containing protrusions were frequently observed. These cells could contain multiple nuclei of varying sizes and had a high content of lysosomes. No significant differences in mitochondrial ultrastructure were observed in nestin-positive and -negative cells, although functional characteristics evaluated by the membrane potential probe differed.

Objectives: Metabolic substrate deficiency is a key factor in many pathologies, with organ vulnerability depending on specialized metabolic profiles. In this study, we aimed to investigate the impact of deprivation stress on mitochondria and cell functions in different cell types and to assess the potential of fumarate, a tricarboxylic acid (TCA) cycle intermediate, to modulate these stress responses. Methods: We assessed the effects of fumarate on cell proliferation and mitochondrial membrane potential under both normal conditions and serum deprivation in vitro in astrocytes, renal epithelial cells, and hepatic stellate cells. Subsequently, we performed bioinformatic analysis of transcriptomic data for brain, kidney, and liver tissues subjected to ischemia to reveal specific patterns of alterations in energy metabolism. Results: Analysis of mitochondria-associated gene expression revealed striking organ-specific differences in transcriptional responses to ischemia, with a significant decrease in expression of genes related to the TCA cycle and electron transport chain in the kidneys and liver, in contrast to the brain. In an in vitro serum deprivation model, fumarate preserved mitochondrial potential in a cell type-specific manner, with optimal concentrations of 12.5 mM for astrocytes (p < 0.05), and 25 mM for renal (p < 0.01) or hepatic cells (p < 0.01). Under normal conditions, fumarate increased mitochondrial potential in astrocytes (p < 0.0001), with weak or no effect on renal and hepatic cells. Estimation of cell number indicates the effects of fumarate on the proliferation of kidney (p < 0.05) and liver (p < 0.0001) cells in normal conditions and cell death after serum deprivation. Conclusion: Bioinformatic analysis demonstrates that ischemic stress induces fundamentally different transcriptional programs in the brain, kidney, and liver, particularly affecting genes involved in mitochondrial bioenergetics. Complementing these findings, our in vitro data identify fumarate as a metabolically active but organ-dependent candidate for tissue protective strategies in substrate deprivation pathologies, like ischemia.

Background/Objectives: Inflammation and oxidative stress are key factors contributing to the initiation and progression of liver fibrosis in chronic obstructive cholestasis. Pantothenic acid (PA) and some of its derivatives have been reported to exhibit moderate anti-inflammatory, antioxidant, and regenerative effects. This study aimed to evaluate the redox-modulating effects of PA derivatives—panthenol (PL), pantethine (PT), and hopantenic acid (HPA) in a rat model of chronic obstructive cholestasis induced by common bile duct ligation (BDL). Methods: Macroscopic, histological, and ultrastructural alterations in the liver were assessed, along with molecular markers of oxidative stress, inflammation, and parameters of the glutathione (GSH) system. Results: BDL-induced liver injury was associated with enhanced lipid peroxidation, mitochondrial structural alterations, depletion of GSH, increased levels of protein S-glutathionylation (PSSG), and elevated thiobarbituric acid-reactive substances in mitochondria. Treatment with PL and, to a lesser extent, PT was associated with attenuation of hepatocellular ultrastructural damage, reduced bile duct hyperplasia, decreased inflammatory and necrotic changes, and moderate improvement in fibrosis-related parameters. In contrast, HPA (a PA antagonist) did not demonstrate hepatoprotective effects and it was associated with more pronounced liver injury. Conclusions: Chronic BDL is accompanied by suppression of glutathione redox capacity and enhanced oxidative stress. PL and PT, but not HPA, were associated with reduced levels of protein S-glutathionylation and partial restoration of redox balance. The protective effects of PL and PT may contribute to their antifibrotic activity, potentially through direct antioxidant capacity or redox-modulating mechanisms associated with the GSH system.

Drug-induced nephrotoxicity is a leading cause of acute kidney injury (AKI) and subsequent chronic kidney disease. Nephrotoxicity often develops as a consequence of treatment with commonly prescribed aminoglycoside antibiotics, and remains a significant clinical challenge. One approach to treating AKI and its associated complications is caloric restriction or its pharmacological mimetics. This study aimed to evaluate the effects of caloric restriction mimetic hydroxycitrate (HC) in gentamicin-induced nephrotoxicity, with particular focus on the influence of treatment duration and the underlying molecular mechanisms. In vitro renal tubular epithelial cells models were used to assess HC’s effects on viability, proliferation, and autophagy activation. For in vivo validation, rats with gentamicin-induced AKI received HC treatment via two distinct regimens (3-week and 7-week administration). Experiments on renal tubule cells showed that HC significantly increased cell viability and proliferation and led to the activation of autophagy. In the rat model, only the 7-week administration of HC demonstrated significantly attenuated renal dysfunction in gentamicin-induced AKI. Moreover, it reduced macrophage infiltration, increased renal cell tolerance to apoptosis, activated autophagy, and reduced oxidative stress. Thus, our results indicate that 7-week HC administration could be used as a prophylactic strategy against antibiotic nephrotoxicity, exerting its effects by promoting autophagy, resisting apoptosis, and attenuating oxidative damage.

We explored the possibility of antioxidant and antifibrotic effects of panthenol (PL) associated with modulation of coenzyme A (CoA) biosynthesis in the liver in a rat model of chronic obstructive cholestasis induced by bile duct ligation (BDL). We found that PL increased alcohol dehydrogenase (ADH) activity in the liver of BDL rats. PL and its analog pantethine increased pantothenate kinase (PANK) activity, restored hepatic CoA levels reduced by BDL, lowered protein-bound CoA, and normalized impaired mitochondrial functions associated with induced oxidative stress after BDL. These effects were accompanied by decreased collagen deposition and improved morphological features of hepatocytes. In contrast, PANK inhibitor, hopantenic acid (HPA), reduced hepatic CoA levels, aggravated hepatocellular damage, and promoted fibrosis. In the human hepatic stellate cell line LX-2, PL exhibited no cytotoxicity over a wide concentration range, increased intracellular CoA levels, decreased reactive oxygen species (ROS) production, and attenuated collagen accumulation associated with oxidative stress in vitro. Importantly, inhibition of ADH by 4-methylpyrazole completely abolished the protective effects of panthenol, indicating that its activity depends on metabolic pathways involving CoA. Notably, PL did not directly reduce H2O2 or superoxide anion radical production in cell-free systems but significantly suppressed lipid peroxidation in liposomes and red blood cells in vitro. Ultimately, these findings indicate that the antioxidant and antifibrotic effects of PL are associated with modulation of CoA metabolism and enhanced resistance of biological membranes to oxidative damage.

Caloric restriction (CR) is known to activate a broad spectrum of cytoprotective signaling pathways and enhance tissue tolerance to various stressors, including those associated with the cytotoxic effects of pharmaceutical agents. Nephrotoxic drugs, such as aminoglycoside antibiotics, remain a major clinical concern due to their frequent use and potential to cause acute kidney injury (AKI), for which effective preventive strategies are still limited. In this study, we investigated whether CR applied for 5 weeks (4-week pretreatment + 1-week concurrent with AKI induction) can alleviate AKI triggered by the antibiotic gentamicin, with a focus on evaluating changes in antioxidant-related parameters and autophagy-associated signaling during CR-mediated nephroprotection. CR’s nephroprotective effects were evaluated using diagnostic assays, Western blotting, and histological analysis. Additionally, oxidative stress markers and mitochondrial integrity were assessed to analyze the impact of CR on antioxidant-related pathways. CR significantly improved renal function and structure, with reduced kidney injury markers (KIM-1, NGAL) and alleviated histological damage. Critically, CR mitigated oxidative stress, evidenced by decreased thiobarbituric acid reactive substances (TBARS) and protein carbonylation, as well as increased levels of the reduced form of glutathione and activity of glutathione peroxidase (GPx). A lowered Bcl-XL/XS ratio was consistent with reduced apoptotic signaling, while reduced leukocyte infiltration reflected attenuated renal inflammation. Additionally, a reduction in mitochondrial DNA (mtDNA) lesions suggested that CR was associated with modulation of mitochondrial and metabolism-related pathways, with concurrent improvements in mitochondrial stability. Our findings demonstrate that CR attenuated gentamicin-induced AKI and was associated with changes in antioxidant-related parameters, reduced mtDNA damage, a decrease in inflammatory cell infiltration, and modulation of autophagy-related signaling.

Diabetes and obesity are associated with poorer outcomes after ischemic stroke; however, it remains unclear whether this results from increased neuronal susceptibility to injury or from vascular dysfunction induced by metabolic syndrome. To minimize the contribution of vascular factors, we used a model of photoinduced thrombosis (PT) in cortical vessels, which generates lesions of reproducible size and is less dependent on collateral blood flow. PT was induced in wild-type (WT) mice, as well as in ob/ob (leptin-deficient) and db/db (leptin receptor-deficient) mice. Magnetic resonance imaging (MRI) revealed that PT produced comparable infarct volumes in all mouse groups. Several genes associated with inflammation and activation of microglia and macroglia in the peri-infarct area (Cst7, Ccl3, Tlr2, Gfap) exhibited similar expression patterns across all three mouse strains, while transcriptional response to cerebral ischemia of Tnfa, Cxcl9, Il6, Cox2, Mmp3, and Bdnf genes depended on the genotype. Overall, despite individual differences in the expression profiles of certain genes, disruption of leptin signaling (whether due to leptin deficiency or leptin receptor deficiency) caused no genotype-specific exacerbation of stroke-induced injury. Assessment of post-stroke neurological deficits revealed substantial differences in absolute scores between WT and ob/ob or db/db mice, attributable to baseline disparities in body weight and motor activity. In db/db mice, normalization of post-stroke neurological status scores to pre-injury values revealed a more pronounced relative functional decline compared to ob/ob mice, suggesting impairments in early compensatory mechanisms and an important role of leptin signaling in neuroplasticity rather than in the extent of acute neuronal damage. Thus, under conditions that minimize vascular complications, neither leptin deficiency nor leptin receptor deficiency exacerbated acute ischemic brain damage or neuroinflammation.

Acute kidney injury (AKI) is a life-threatening clinical syndrome marked by a rapid decline in renal function, contributing to substantial morbidity and mortality. Emerging evidence suggests that modulating macrophage polarization represents a promising therapeutic strategy for AKI. However, achieving precise pharmacological regulation remains a challenge. To address this, we developed a novel macrophage-targeting nanoplatform (GMO@GO@PEG@MAN) by loading germacrone onto mannose-functionalized graphene oxide nanosheets. This engineered system enhances renal accumulation and enables macrophage-specific delivery in AKI models, while significantly suppressing pro-inflammatory M1 macrophage polarization. It provides multi-mechanistic renal protection by attenuating oxidative stress, promoting NRF2 nuclear translocation, inhibiting ferroptosis, and exerting potent broad-spectrum antimicrobial activity against both extracellular and intracellular pathogens. Our findings establish this nanoplatform as an effective multi-targeted therapy for AKI, demonstrating robust efficacy across both in vitro and in vivo models. This synergistic therapeutic strategy-combining macrophage reprogramming, oxidative stress mitigation, and antimicrobial activity-marks a significant advancement in AKI treatment paradigms.

Ischemia–reperfusion (I/R) injury is a complex pathological process underlying numerous acute organ failures and is a significant cause of morbidity and mortality in diseases such as myocardial infarction, stroke, thrombosis, and organ transplantation. Mesenchymal stem cell (MSC)-derived extracellular vesicles (EVs) have demonstrated considerable therapeutic potential, but their broad tropism and general repair signaling may limit their efficacy. This review addresses the emerging paradigm of using organ-specific EVs for the treatment of I/R injury in the respective organs. We summarize the existing studies performed on experimental animals showing that these native EVs could possess tissue tropism and carry a specialized cargo of proteins, miRNAs, and lipids tailored to the unique regenerative needs of their organ of origin, enabling them to precisely modulate key processes, including inflammation, apoptosis, oxidative stress, and angiogenesis. However, their clinical translation faces challenges related to scalable production, standardization, and the dualistic nature of their effects, which can be either protective or detrimental, depending on the cellular source and pathophysiological context. Future developments need to focus on overcoming these obstacles through rigorous isolation protocols, engineering strategies such as cargo enrichment and hybrid vesicle creation, and validation in large-animal models. Overall, organ-specific EVs offer a novel, cell-free therapeutic strategy with the potential to significantly improve outcomes in I/R injury.

Background: Mitochondrial dysfunction plays a critical role in the pathogenesis of Alzheimer's disease (AD). Resveratrol is a promising compound for the treatment of various neurodegenerative diseases, including AD. Aims: To investigate mitochondrial damage and the effects of resveratrol on inflammation, cognitive function, and mitochondrial quality control in APP/PS1 mice. Methods: Comparative analysis of mitochondrial DNA (mtDNA) damage was conducted between 10-month-old APP/PS1 mice and age-matched C57BL/6 mice. Assessments included measurement of amyloid-beta levels, inflammatory markers, swimming distance in the Morris water maze, and gut microbiome composition. Resveratrol's effects on cytokine expression, mtDNA levels in plasma, and activation of Nuclear factor erythroid 2-related factor 2/Antioxidant response element (Nrf2/ARE) and phosphoinositide 3-kinase/protein kinase B (also known as Akt)/mechanistic target of rapamycin complex 1 (PI3K/Akt/mTORC1) signaling pathways were also evaluated. Results: APP/PS1 mice exhibited significantly increased mtDNA damage in the prefrontal cortex, midbrain, and cerebellum, alongside higher amyloid-beta levels and inflammatory markers. Resveratrol treatment led to reduced expression of pro-inflammatory cytokines, a decrease in Proteobacteria levels, and lower cellfree mtDNA in plasma. Partial improvement in long-term spatial memory was observed in APP/PS1 mice following resveratrol treatment, likely due to its anti-inflammatory properties. Activation of the Nrf2/ARE signaling pathway and markers of PI3K/Akt/mTORC1 axis activation were noted, with the latter regulating long-term potentiation. Conclusion: Resveratrol demonstrates potential in mitigating inflammation and improving mitochondrial quality control in APP/PS1 mice, but it does not reduce amyloid-beta levels, highlighting the complexity of AD pathology and the need for further research.

The sarcomeric giant protein titin affects the passive elasticity of the heart muscle and is crucial for proper cardiac function, including diastolic relaxation of the left ventricle. A useful common method for studying titin is electrophoretic analysis which can be used to examine the distribution of its isoforms in the heart. There are 5 titin parameters that can be analyzed: the N2BA/N2B isoforms ratio, the T2/T1 bands ratio, Cronos isoform content, NT isoform content, the total titin-to-myosin heavy chain (TT/MHC) ratio. These parameters can only be assessed through electrophoresis of giant proteins. It is known that these parameters are related to various biomolecular processes in muscle cells, such as providing of elastic properties, turnover, contraction, and maintaining a highly ordered sarcomere structure. In this review, we discuss the diagnostic potential of electrophoretic visualization of cardiac titin changes in various human heart diseases and animal models of physiological adaptations or pathologies.

Traumatic brain injuries (TBIs) are a serious problem affecting individuals of all ages. Mitochondrial dysfunctions represent a significant form of secondary injury and may serve as a promising target for therapeutic intervention. Our research demonstrated that craniotomy, which precedes the experimental induction of trauma in mice, can cause considerable damage to mitochondrial DNA (mtDNA), disrupt the regulatory expression of angiogenesis, and increase inflammation. However, the reduction in the mtDNA copy number and glial activation occur only after a direct impact to the brain. We explored two potential therapeutic agents: the dietary supplement L-carnitine—a potential reserve source of ATP for the brain—and the cardiac drug mildronate, which inhibits L-carnitine but activates alternative compensatory pathways for the brain to adapt to metabolic disturbances. We found that L-carnitine injections could protect against mtDNA depletion by promoting mitochondrial biogenesis. However, they also appeared to aggravate inflammatory responses, likely due to changes in the composition of the gut microbiome. On the other hand, mildronate enhanced the expression of genes related to angiogenesis while also reducing local and systemic inflammation. Therefore, both compounds, despite their opposing metabolic effects, have the potential to be used in the treatment of secondary injuries caused by TBI.

Renocardiac syndrome type 4 (RCS4) is a common comorbid pathology, but the mechanisms of kidney dysfunction-induced cardiac remodeling and the involvement of cardiac progenitor cells (CPCs) in this process remain unclear. The aim of this study was to investigate the structural and functional changes in the cardiac muscle in RCS4 induced by unilateral ureteral obstruction (UUO) and the role of nestin+ CPCs in these. Heart function and localization of nestin+ cells in the myocardium were assessed using nestin-GFP transgenic mice subjected to UUO for 14 and 28 days. UUO resulted in cardiac hypertrophy, accompanied by an elongation of the QRS wave on the ECG, decreased expression of Cxcl1, Cxcl9, and Il1b, reduced the number of CD11b+ cells, and increased in titin isoform parameters, such as T1/MHC and TT/MHC ratios, without changes in fibrosis markers. The number of nestin+ cells increased in the myocardium with increased duration of UUO and displayed an SCA-1+TBX5+ phenotype, consistent with CPCs. Thus, cardiac pathology in RCS4 was manifested by cardiomyocyte hypertrophy with changes in the electrophysiological phenotype of the heart, not accompanied by fibrosis or inflammation. Nestin+ cardiac cells retained the CPC phenotype during UUO, and their number increased, which suggests their participation in regenerative processes in the heart.

The levels of inflammatory markers increased in both mouse blood plasma and affected brain area 24 days after traumatic brain injury, which was accompanied by impairment of spatial working memory. Methylene blue administered during the first 3 days after injury reduced the levels of some inflammation markers and increased the expression of genes involved in the regulation of mitochondrial biogenesis and mitophagy, i.e. genes responsible for mitochondrial quality control. Additionally, methylene blue partially mitigated the cognitive deficits induced by the injury, suggesting it as a promising compound for maintaining brain function after traumas.

Acute cerebral ischemia caused by stroke,traumatic brain injury(TBI),or systemic acute conditions such as hemorrhagic shock,cardiac arrest,or disseminated intravascular coagulation results in an energy crisis in local sites or the whole brain.The disruption of cerebral blood flow deprives the brain cells of oxygen and glucose,the essential substrates for adenosine triphosphate(ATP)synthesis.As a result,oxidative phosphorylation in the mitochondria fails,forcing cells to rely on anaerobic glycolysis(He et al.,2020).Although this compensatory mechanism maintains short-term energy production under hypoxic conditions,overall ATP production is significantly reduced.Neurons,which are highly susceptible to ischemic injury,deplete their ATP stores faster than glial cells(e.g.,astrocytes),which have some energy reserves.

Severe injuries and some pathologies associated with massive bleeding, such as maternal hemorrhage, gastrointestinal and perioperative bleeding, and rupture of an aneurysm, often lead to major blood loss and the development of hemorrhagic shock. A sharp decrease in circulating blood volume triggers a vicious cycle of vasoconstriction and coagulopathy leading to ischemia of all internal organs and, in severe decompensated states, ischemia of the brain and heart. The basis of tissue damage and dysfunction in hemorrhagic shock is an interruption in the supply of oxygen and substrates for energy production to the cells, making the mitochondria a source and target of oxidative stress and proapoptotic signaling. Based on these mechanisms, different strategies are proposed to treat the multiple organ failure that occurs in shock. The main direction of such treatment is to provide the cells with a sufficient amount of substrates that utilize oxidative phosphorylation at different stages and increase the efficiency of energy production by the mitochondria. These strategies include restoring the efficiency of mitochondrial complexes, for example, by restoring the nicotinamide adenine dinucleotide (NAD) pool. Another direction is approaches to minimize oxidative stress as well as apoptosis, which are primarily dependent on the mitochondria. There are also a number of other methods to reduce mitochondrial dysfunction and improve the quality of the mitochondrial population. In this review, we consider such strategies for the treatment of hemorrhagic shock and show the promise of therapeutic approaches aimed at restoring the bioenergetic functions of the cell and protecting mitochondria.