Renal ischemia-reperfusion injury (RIRI) is a common cause of acute renal injury. Studies have shown that sodium aescinate (SA) may serve as a potential therapeutic agent, although its exact mechanism remains unclear. This study first evaluated the efficacy of SA using a mouse renal ischemia-reperfusion model. Subsequently, its mechanism was elucidated through systematic bioinformatics, and finally validated through in vitro and in vivo experiments. The results demonstrated that SA has a protective effect on renal function in mice with RIRI. Bioinformatic analysis indicated that the pyroptosis pathway is significantly activated during renal ischemia-reperfusion injury, and immunohistochemistry showed that the level of renal pyroptosis is upregulated during ischemia-reperfusion injury. Administration of SA was able to reduce the expression of pyroptosis-related proteins (GSDMD, NLRP3, IL-1β) in RIRI. In vitro and in vivo experiments further confirmed that SA exerts an anti-pyroptotic effect by inhibiting the AKT/NLRP3 signaling pathway. Ultimately, SA mitigates kidney injury in IRI mice by suppressing renal failure through inhibition of the AKT/NLRP3 signaling pathway.

JOURNAL/nrgr/04.03/01300535-202509000-00022/figure1/v/2024-11-05T132919Z/r/image-tiff Poststroke cognitive impairment is a major secondary effect of ischemic stroke in many patients; however, few options are available for the early diagnosis and treatment of this condition. The aims of this study were to (1) determine the specific relationship between hypoxic and α-synuclein during the occur of poststroke cognitive impairment and (2) assess whether the serum phosphorylated α-synuclein level can be used as a biomarker for poststroke cognitive impairment. We found that the phosphorylated α-synuclein level was significantly increased and showed pathological aggregation around the cerebral infarct area in a mouse model of ischemic stroke. In addition, neuronal α-synuclein phosphorylation and aggregation were observed in the brain tissue of mice subjected to chronic hypoxia, suggesting that hypoxia is the underlying cause of α-synuclein-mediated pathology in the brains of mice with ischemic stroke. Serum phosphorylated α-synuclein levels in patients with ischemic stroke were significantly lower than those in healthy subjects, and were positively correlated with cognition levels in patients with ischemic stroke. Furthermore, a decrease in serum high-density lipoprotein levels in stroke patients was significantly correlated with a decrease in phosphorylated α-synuclein levels. Although ischemic stroke mice did not show significant cognitive impairment or disrupted lipid metabolism 14 days after injury, some of them exhibited decreased cognitive function and reduced phosphorylated α-synuclein levels. Taken together, our results suggest that serum phosphorylated α-synuclein is a potential biomarker for poststroke cognitive impairment.

Aging is characterized by a steady loss of physiological integrity, leading to impaired function and increased vulnerability to death. Cell senescence is a biological process that progresses with aging and is believed to be a key driver of age-related diseases. Senescence, a hallmark of aging, also demonstrates its beneficial physiological aspects as an anti-cancer, pro-regenerative, homeostatic, and developmental mechanism. A transitory response in which the senescent cells are quickly formed and cleared may promote tissue regeneration and organismal fitness. At the same time, senescence-related secretory phenotypes associated with extended senescence can have devastating effects. The fact that the interaction between senescent cells and their surroundings is very context-dependent may also help to explain this seemingly opposing pleiotropic function. Further, mitochondrial dysfunction is an often-unappreciated hallmark of cellular senescence and figures prominently in multiple feedback loops that induce and maintain the senescent phenotype. This review summarizes the mechanism of cellular senescence and the significance of acute senescence. We concisely introduced the context-dependent role of senescent cells and SASP, aspects of mitochondrial biology altered in the senescent cells, and their impact on the senescent phenotype. Finally, we conclude with recent therapeutic advancements targeting cellular senescence, focusing on acute injuries and age-associated diseases. Collectively, these insights provide a future roadmap for the role of senescence in organismal fitness and life span extension.

Ischemic heart disease (IHD) is a leading cause of morbidity and mortality worldwide. Myocardial ischemia/reperfusion injury (MIRI) is the primary cause of myocardial injury triggered by post-myocardial infarction reperfusion therapy. Its pathogenesis involves Ca2+ overload, the production of large amounts of oxygen-free radicals, inflammation, and cell necrosis. Growing evidence suggests that the NLRP3 inflammasome significantly contributes to the sterile inflammatory response and pyroptosis in MIRI, linking damage sensing to the initiation and amplification of the inflammatory response. Reportedly, ozone exerts anti-inflammatory and anti-infection effects by activating the antioxidant system. Additional evidence suggests that ozone inhibits NLRP3 inflammasome expression to relieve ischemic injury. In this study, we aimed to explore whether pretreating the myocardium with ozone protects it from MIRI by inhibiting the NLRP3 inflammasome. Rats were subjected to rectal infusion of ozone for 5 consecutive days, followed by ligation of the left anterior descending coronary artery for 30 min and reperfusion for 120 min to induce MIRI. Experimental results were obtained using echocardiography, triphenyltetrazolium chloride and hematoxylin and eosin staining, western blotting, and enzyme-linked immunosorbent assay. The results showed that ozone significantly improved the diastolic function of the heart, reduced the area of myocardial infarction, and decreased the expression levels of NLRP3, pro-caspase-1, ASC, and the secretion of caspase-1, interleukin (IL)-1β, and IL-18. In summary, these findings reveal that ozone pretreatment can alleviate the damage that occurs during MIRI by inhibiting the NLRP3 Inflammasome.

Sickle cell diseases are widespread in regions encompassing the Mediterranean, Middle East, sub-Saharan Africa, and specific parts of Asia, primarily due to the abnormal production of hemoglobin S. This genetic blood disorder stems from a mutation in the beta-globin gene, a crucial component of hemoglobin and the heme-containing protein found in red blood cells. Point mutations in the hemoglobin gene can be inherited as a heterozygous or homozygous pattern. These mutations disrupt the normal configuration of the protein, impeding its physiological function and altering the cell's shape, giving it a sickle-like appearance. The resulting sickle cells can lead to organ damage, intense physical discomfort, and anemia; in severe cases, the condition can be fatal. Early detection and effective treatment methods have the potential to progressively reduce the associated mortality rate over time. To diagnose sickle cell disease and its carrier states with unparalleled specificity, a variety of approaches have been developed. The most common method includes differential blood cell counts and their assessment, high-performance liquid chromatography (HPLC) and hemoglobin electrophoresis. Furthermore, innovative sensing technologies are currently under development, encompassing user-friendly, cost-effective and portable point-of-care devices that are capable of timely diagnosis at the genetic and molecular levels of these disorders. The review delves into a range of established and innovative strategies utilized in the detection of sickle cell disease, also underscoring the essential role played by diverse bio-sensing techniques in propelling the advancement of early diagnosis of SCD.

Hepatic ischemia/reperfusion (I/R) injury (HIRI) is an intrinsic phenomenon observed in the process of various liver surgeries. Unfortunately, there are currently few options available to prevent HIRI. Accordingly, we aim to explore the role and key downstream effects of B-cell lymphoma 6 (BCL6) in hepatic I/R (HIR).

BCL6 expression levels were measured in I/R liver tissue and primary hepatocytes stimulated by hypoxia/reoxygenation (H/R). Moreover, we ascertained the BCL6 effect on HIR in vivo using liver-specific BCL6 knockout mice and adenovirus-BCL6-infected mice. RNA-sequencing, luciferase, chromatin immunoprecipitation, and interactome analysis were combined to identify the direct target and corresponding molecular events contributing to BCL6 function. DNA pull-down was applied to identify upstream of BCL6 in the H/R challenge.

HIR represses BCL6 expression in vivo and in vitro. Hepatic BCL6 overexpression attenuates inflammation and apoptosis after I/R injury, whereas BCL6 deficiency aggravates I/R-induced liver injury. RNA-sequencing showed that BCL6 modulated nucleotide-binding oligomerization domain, leucine-rich repeat and pyrin domain-containing 3 inflammasome signaling in HIRI. Mechanistically, BCL6 deacetylated nuclear factor kappa-B p65 lysine 310 by recruiting sirtuin 1 (SIRT1), thereby inhibiting the nuclear factor kappa-B/nucleotide-binding oligomerization domain, leucine-rich repeat and pyrin domain-containing 3 pathway. Moreover, overexpression of SIRT1 blocked the detrimental effects of BCL6 depletion. Moreover, EX 527, a SIRT1 inhibitor, vanished protection from BCL6 overexpression. Furthermore, transcription factor 7 was found to mediate the transcription regulation of BCL6 on H/R challenge.

Our results provide the first evidence supporting BCL6 as an important protective agent of HIR. This suggests a potential therapeutic approach for HIR.

Ischemic-related diseases, such as myocardial infarction and stroke, are primarily driven by a deficit in oxygen supply leading to cellular damage and death. Ferroptosis has emerged as an important mechanism contributing to the progression of ischemic injury, characterized by iron-dependent lipid peroxidation. This review aims to provide a comprehensive overview of the significant mechanisms underlying ferroptosis in ischemic conditions and explores the potential effects of traditional Chinese medicines (TCMs) and their extracts. Numerous compounds extracted from TCMs, including flavonoids, polyphenols and terpenes, exhibit potent antiferroptotic effects by activating nuclear factor erythroid 2-related factor 2, upregulating glutathione peroxidase 4, inhibiting lipid peroxidation and so on. These properties render TCMs a promising candidate for developing novel ferroptosis therapeutic strategies. This review underscores the importance of investigating the interactions between ferroptosis and TCMs within the context of ischemic diseases. These findings provide valuable insights for future research to identify targets associated with ferroptosis regulation, thereby expanding the pharmacological perspective of TCMs in treating ischemic diseases and revealing the potential of novel therapeutic strategies. Additionally, this highlights the relevance of integrating traditional and modern medical approaches in addressing complex health issues.

Stroke is the leading cause of physical disability and death among adults in most Western countries. Consecutive to a vascular occlusion, cells face a brutal reduction in supply of oxygen and glucose and thus an energy failure, which in turn triggers cell death mechanisms. Among brain cells, neurons are the most susceptible to ischemia because of their high metabolic demand and low reservoir of energy substrates. In neurons, glycolysis uses glucose coming from blood or from glycogen stored in astrocytes, underlying the deep astrocyte-neuron metabolic cooperation. During ischemia, both the aerobic and anaerobic pathways and thus energy production are compromised, which disrupts proper cell functioning, notably Na+/K+ ATPase and mitochondria. This results in altered Ca2+ homeostasis and overproduction of ROS, the latter being further exacerbated during the reperfusion phase. Consequently, glucose metabolism in the different brain cell populations plays a central role in injury and recovery after stroke, and has recently emerged as a promising target for therapeutic intervention. In this context, the overall objective of this article is to review the interconnections between stroke and brain glucose metabolism and to explore how its targeting may offer new therapeutic opportunities in addressing the global stroke epidemic.

Hepatic ischemia-reperfusion injury (HIRI) is a common perioperative complication occurring after liver transplantation and can lead to further problems such as early allograft dysfunction (EAD). Currently, the treatment options for HIRI are extremely limited. In this study, we used bioinformatics analysis to elucidate the critical role of neutrophil chemokines (CXC chemokines) in HIRI. By analyzing sequencing data from the hepatic tissue of posttransplant patients with EAD and the reperfused animal model, we discovered that hepatocytes and macrophages are the primary cells secreting CXC chemokines, and the activation of the nuclear factor kappa B (NF-κB) signaling pathway is the main driver of their secretion. Melatonin (MT) can protect cells from oxidative harm while also inhibiting NF-κB signaling, suggesting its potential to ameliorate HIRI. Accordingly, we designed a nanoparticle platform coated with genetically engineered macrophage membranes-called CXCR2-MM@PLGA/MT-to target the cells secreting CXC chemokines. CXCR2 overexpression on the macrophage membranes not only enhanced the targeting capacity of the nanoparticles but also prevented neutrophil infiltration via the scavenging of CXC chemokines. Meanwhile, the MT delivered to the site of injury successfully attenuated CXC chemokine release after macrophage polarization and hepatocyte necrosis by inhibiting NF-κB phosphorylation and inducing antioxidant effects. Through the synergistic effects of MT and the CXCL/CXCR axis-blocking function of the engineered nanoparticles, CXCR2-MM@PLGA/MT attenuated the aggregation of neutrophils at the site of injury, markedly reducing local inflammation and cellular damage following HIRI. This engineered cellular nanoparticle-based therapy could thus serve as a safe, effective, and cost-efficient strategy for treating HIRI.

The aim of this study was to determine if remote ischemic per-conditioning (RIPerC) can provide protection to the kidneys from ischemia-reperfusion injury (IRI) by increasing the expression of the Gclc and Gclm genes involved in innate defenses. Rats undergoing sham surgery were used as controls. Induction of renal IRI involved blocking the renal pedicles for 60 min, then allowing 24 h of reperfusion. RIPerC involved 4 cycles (5 min) of limb I/R. Animals were divided into seven groups in a random manner: sham, I/R, I/R + RIPerC, I/R + NaHS (NaHS, 100 μmol /kg, i.p), I/R + RIPerC+NaHS, I/R + PAG (propargyl glycine, 50 mg/kg, i.p.) and I/R + RIPerC+PAG. Following reperfusion, samples of urine, blood, and renal tissue were gathered for functional, molecular, and histological analysis. Renal IRI impaired kidney function (reduced CCr, increased FENa, decreased water reabsorption, and reduced urine osmolality), increased oxidative stress (an increase in total oxidative status and a decrease in total antioxidant capacity), and reduced expression of CBS, CSE, Gclc and Gclm genes, causing tissue damage. RIPerC attenuated the IRI-induced kidney dysfunction, oxidative stress, and gene expression changes. Inhibiting hydrogen sulfide signaling with propargylglycine reduced the benefits of RIPerC, while the hydrogen sulfide donor NaHS enhanced them. These findings suggest RIPerC's renal protective effects involve upregulation of antioxidant defense pathways.

The causal relationship between the activation of nuclear factor erythroid 2-related factor 2 (NRF2) and the preservation of SERCA2a function in mitigating myocardial ischemia-reperfusion (mI/R) injury, along with the associated regulatory mechanisms, remains incompletely understood. This study aims to unravel how NRF2 directly or indirectly influences SERCA2a function and its regulators, phospholamban (PLN) and Dwarf Open Reading Frame (DWORF), by testing the pharmacological repositioning of AEOL-10150 (AEOL) in the context of mI/R injury.

C57BL6/J, Nrf2 knockout (Nrf2-/-), and wild-type (Nrf2+/+) mice, as well as human induced pluripotent stem cell-derived cardiomyocytes (hiPSCMs) were subjected to I/R injury. Gain/loss of function techniques, RT-qPCR, western blotting, LC/MS/MS, and fluorescence spectroscopy were utilized. Cardiac dimensions and function were assessed by echocardiography.

In the early stages of mI/R injury, AEOL administration reduced mitochondrial ROS production, decreased myocardial infarct size, and improved cardiac function. These effects were due to NRF2 activation, leading to the overexpression of the micro-peptide DWORF, consequently enhancing SERCA2a activity. The cardioprotective effect induced by AEOL was diminished in Nrf2-/- mice and in Nrf2/Dworf knockdown models in hiPSCMs subjected to simulated I/R injury. Our data show that AEOL-induced NRF2-mediated upregulation of DWORF disrupts the phospholamban-SERCA2a interaction, leading to enhanced SERCA2a activation and improved cardiac function.

Taken together, our study reveals that AEOL-induced NRF2-mediated overexpression of DWORF enhances myocardial function through the activation of the SERCA2a offering promising therapeutic avenues for mI/R injury.

Sickle cell disease (SCD) is a genetic disorder characterized by sickle red blood cells (RBCs). Sickle RBCs cause cerebral vasculopathies including vaso-occlusive events, leading to ischemia-reperfusion injury and hypoxic tissue environment. To date, the physiological blood flow velocities in cerebral vessels of preclinical SCD models has not been evaluated under hypoxic-reoxygenation. In our study, we used transcranial ultrasound techniques to measure abnormal blood flow velocities in the internal carotid (ICA) and middle cerebral arteries (MCA) of transgenic sickle cell mice (SS) challenged with hypoxia-reoxygenation. Our study showed that SS mice that underwent hypoxic stress exhibited lower relative mean velocities in the MCA compared to wildtype mice (AA) (0.67 ± 0.18 vs. 0.95 ± 0.15; p < 0.05). Comparison of the Lindegaard ratio between normoxia and hypoxia in SS mice suggested that the MCA underwent vasodilation (0.67 ± 0.18 vs. 0.95 ± 0.15; p < 0.05). Bilirubin, a potential biomarker for cerebral vasculopathies in SCD, was higher in SS than AA mice (0.56  ±   0.28 vs. 0.05  ±   0.07 mg/dL; p < 0.05). Correlation analyses revealed a significant association between bilirubin levels and blood velocities of MCA (r = -0.9377, p = 0.0002) and ICA (r = 0.8203, p = 0.0068), especially in hypoxic conditions of SS mice. We propose that the reactivity of cerebral vessels in SS mice is correlated with the elevated plasma bilirubin level.

Cerebral ischemia-reperfusion (I/R) is a condition that occurs when blood flow is restored after a temporary interruption and may lead to deterioration in brain functions depending on the time passed. One of the changes in functions is neurological score values. This study aimed to determine the effect of brain ischemia reperfusion and 2-week naringin supplementation on changes in neurological score and neurogenesis in ovariectomized female rats. Experimental groups of 36 Wistar-albino-type female rats were created as follows: control group: no anesthesia or surgical procedure was applied. Ovariectomy-sham brain I/R group: After the ovariectomy was performed under general anesthesia, the carotid artery regions were opened and closed, and sham ischemia-reperfusion was performed, followed by a vehicle application for 2 weeks (2 weeks, 1 ml 0.25% carboxymethylcellulose). Ovariectomy-I/R group: After ovariectomy, carotid arteries were isolated under general anesthesia, ligated for 30 min, and reperfused for 2 weeks after ischemia was performed. Ovariectomy-I/R sham treatment group: After ovariectomy, the carotid arteries were isolated under general anesthesia, then ligated and ischemia was performed for 30 min, and then reperfusion and vehicle application were performed for 2 weeks. Ovariectomy-I/R naringin treatment group: After ovariectomy, carotid arteries were isolated under general anesthesia, ligated for 30 min, and ischemia was performed, followed by naringin application with reperfusion for 2 weeks. Neurological scoring values performed on the 1st, 7th, and 14th days after the surgical procedure significantly increased with ischemia-reperfusion. Also, hippocampus and frontal cortex calbindin, alpha/beta-tubulin, and Neu-N levels were reduced considerably by ischemia-reperfusion. However, it was observed that a 2-week naringin application significantly suppressed the increase in neurological scores. The suppression in neurological score values became more evident in the 2nd week. Our results show that the impairment of motor functions and neurogenesis in the frontal cortex and hippocampus in brain ischemia-reperfusion after ovariectomy in female rats was significantly improved by 2 weeks of naringin supplementation.

In recent years, metabolite identification of chemical constituents of traditional Chinese medicine (TCM) has been extensively studied. However, due to the intricacy of metabolic processes and the low concentration of metabolites, identifying metabolites of TCM in vivo is still a tough work. Meanwhile, credibility of metabolite identification through mass spectrum technology has been called into question by reason of the lack of metabolite standards. In this study, ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS) was used to detect biological samples including plasma, feces, urine, liver, kidney, brain of normal and middle cerebral artery occlusion (MCAO) rats orally administrated water extract of Danshen-Honghua herbal pair (DHHP). An analysis strategy which combined MS data analysis platform UNIFI with quantitative structure-retention relationship (QSRR) model was established. First, metabolites of DHHP were identified rapidly by utilizing UNIFI analysis platform to analyze acquired MS data. Then, quantitative structure-retention relationships model was built through BP neural network optimized by the ant colony algorithm. Finally, predicted retention times of identified metabolites were produced by QSRR model. Metabolites identified by UNIFI whose difference between predicted and experimental retention time was beyond 1 min were considered false positive and excluded to improve the credibility of identification. According to the established analysis strategy, 26 prototypes and 16 metabolites were identified. Established MS data analysis strategy which combined UNIFI analysis platform with QSRR model was proven to be a creditable method to identify the in vivo metabolites of TCM rapidly and accurately.

Renal ischemia-reperfusion injury (IRI), a common complication following kidney transplantation and partial nephrectomy, is the leading cause of renal dysfunction with limited treatment. Excessive cellular iron accumulation drives lipid peroxidation and activates pathways associated with ferroptosis, which has been implicated in renal IRI. However, the regulatory mechanisms of cellular iron metabolism and its relationship with ferroptosis during ischemia-reperfusion (IR) remain unclear. In this study, in vitro OGSD-R (oxygen, glucose, and serum deprivation-restoration) models and in vivo IR models were employed to investigate alterations in iron metabolism, ferroptosis, and the underlying molecular mechanisms using immunofluorescence, immunoblotting and biochemical testing. We identified glucose deprivation-restoration (GD-R) as a key trigger of cellular iron overload under renal IR condition. Mechanistically, GD-R-induced iron overload is driven by the dysfunction of vacuolar ATPase (V-ATPase)-mammalian target of rapamycin (mTOR) pathway. Inactivation of mTOR results in lysosomal iron releases via two-pore channel 2 (TPC2) and ferritin degradation through ferritinophagy. This process elevates intracellular iron levels, thereby promoting ferroptosis in renal IRI. Targeting cellular iron metabolism effectively alleviates renal IRI. These findings highlight the critical role of glucose metabolism and V-ATPase-mTOR pathway in the regulation of iron homeostasis and ferroptosis during renal IRI, and establish a mechanistic link among glucose metabolism, iron overload and ferroptosis, providing potential therapeutic targets for renal IRI.

Ischemia-reperfusion injury (IRI) causes vascular endothelial dysfunction. Preclinical data suggest that the SGLT2 inhibitor dapagliflozin may protect against vascular IRI. This trial has investigated if oral treatment with dapagliflozin can mitigate the transient impairment of IRI-induced-endothelial dysfunction in the forearm resistance vasculature. 32 healthy males (n = 16 per group, age: 27 ± 4 yrs) were studied in this randomized, placebo-controlled, parallel-group, double-blinded trial. Acetylcholine (ACh; endothelium-dependent vasodilator) and glyceryltrinitrate (GTN; endothelium-independent vasodilator) were administered into the brachial artery of the non-dominant arm. The response to stepwise increasing doses on forearm blood flow (FBF) was assessed. FBF was measured before and after a cuff-induced 20-minute forearm ischemia at pre-dose and following daily intake of 10 mg dapagliflozin or placebo over 15 days. IRI reduced endothelium-dependent vasodilatation by 29% (p < 0.001, paired t-test). After a 15-day treatment period, IRI-induced endothelial dysfunction was abrogated in participants receiving dapagliflozin (FBF AChAUC ratios post- vs. pre-ischemia: dapagliflozin: 0.93; 95% CI: 0.80-1.29) but unchanged with placebo (0.81; 95% CI: 0.68-0.92; p = 0.015 vs. pre-ischemia). GTN-induced vasodilation was not altered by IRI or treatment. Dapagliflozin treatment at standard clinical doses over 15 days prevents IRI-induced vascular endothelial dysfunction in the forearm resistance vasculature of healthy young males. The underlying mechanism and the potential clinical impact remain to be demonstrated.Clinical trial registration https://clinicaltrials.gov/study/NCT05217654 NCT05217654; EudraCT number: 2021-005002-95 Date of registration: 20/01/2022.

Experimental stroke models in rodents are essential for mechanistic studies and therapeutic development. However, these models have several limitations negatively impacting their translational relevance. Here we aimed to develop a minimally invasive thrombotic stroke model through magnetic particle delivery that does not require craniotomy, is amenable to reperfusion therapy, can be combined with in vivo imaging modalities, and can be performed in awake mice. We found that the model results in reproducible cortical infarcts within the middle cerebral artery (MCA) territory with cytologic and immune changes similar to that observed with more invasive distal MCA occlusion models. Importantly, the injury produced by the model was ameliorated by tissue plasminogen activator (tPA) administration. We also show that MCA occlusion in awake animals results in bigger ischemic lesions independent of day/night cycle. Magnetic particle delivery had no overt effects on physiologic parameters and systemic immune biomarkers. In conclusion, we developed a novel stroke model in mice that fulfills many requirements for modeling human stroke.

Background: Intestinal ischemia reperfusion injury (IRI) is a harmful process that occurs during intestinal infarction and intestinal transplantation (ITx). It is characterized by severe inflammation which disrupts the mucosal barrier, causing bacterial translocation and sepsis. Tranilast (N-[3,4-dimethoxycinnamoyl]-anthranilic acid) (TL) is a synthetic compound with powerful anti-inflammatory properties. Objective: To investigate the effect of pretreatment with TL in a validated rat model of intestinal IRI (60 min of ischemia). Methods: TL (650 mg/kg) was administered by oral gavage 24 and 2 h before the onset of ischemia. Experiment 1 examined 7-day survival in 3 study groups (sham, vehicle+IRI and TL+IRI, n = 10/group). In Experiment 2, the effects on the intestinal wall integrity and inflammation were studied after 60 min of reperfusion using 3 groups (sham, IRI and TL+IRI, n = 6/group). The following end-points were studied: L-lactate, intestinal fatty acid-binding protein (I-FABP), histology, intestinal permeability, endotoxin translocation, pro- and anti-inflammatory cytokines and heme oxygenase-1 (HO-1) levels. Experiment 3 examined the role of HO-1 upregulation in TL pretreatment, by blocking its expression using Zinc protoporphyrin (ZnPP) at 20 mg/kg vs. placebo (n = 6/group). Results: Intestinal IRI resulted in severe damage of the intestinal wall and a 10% 7-day survival. These alterations led to endotoxin translocation and upregulation of pro-inflammatory cytokines. TL pretreatment improved survival up to 50%, significantly reduced inflammation and protected the intestinal barrier. The HO-1 inhibitor ZnPP, abolished the protective effect of TL. Conclusions: TL pretreatment improves survival by protecting the intestinal barrier function, decreasing inflammation and endotoxin translocation, through upregulation of HO-1.This rat study of severe intestinal ischemia reperfusion injury demonstrates a novel role for Tranilast as a potential therapy. Administration of Tranilast led to a marked reduction in mortality, inflammation and intestinal permeability and damage. The study proved that Tranilast functions through upregulation of heme oxygenase-1.

Ischemia-reperfusion injury stands as a primary instigator of acute kidney injury (AKI), prominently driven by oxidative stress. Among the critical antioxidant defenses is glutathione peroxidase 3 (GPX3), an enzyme generated by renal tubular epithelial cells. Our prior investigations have unveiled a substantial downregulation of GPX3 in renal tissues gleaned from AKI patients and murine models. This study aims to investigate the role of tubular cell-specific Gpx3 deletion on ischemia-reperfusion injury-induced AKI (IRI-AKI) in a murine model and delineate the potential underlying mechanisms. By generating renal tubular epithelial cell-specific Gpx3 knockout mice and inducing IRI-AKI, we assessed a spectrum of kidney injury indices including renal function, oxidative stress, apoptosis and mitochondrial dynamics. Additionally, we conducted transcriptome sequencing and bioinformatics analyses. The outcomes underscore that the deficiency of GPX3 in tubular cells exacerbates tubular injury, renal dysfunction, oxidative stress, apoptosis, and mitochondrial dynamic disturbances in the context of IRI-AKI. Sequencing and bioinformatics analysis suggest that the Gpx3 deletion predominantly impacts pathways associated with metabolism and inflammation. In conclusion, the tubular cell-specific deficiency of GPX3 exacerbates renal injury by intensifying oxidative stress, fostering mitochondrial impairment, perturbing metabolic processes and fueling inflammation. The targeted restoration of GPX3 in the renal tubular emerges as a potential therapeutic avenue for mitigating IRI-AKI.

Myocardial ischemia/reperfusion (I/R) injury is one of the problems after coronary artery recanalization in patients with acute myocardial infarction, and the discovery of exosomes presents a broad potential for treating myocardial I/R injury. This work examined the function and regulatory mechanisms of exosomes produced from bone marrow mesenchymal stem cells (BMSCs-Exo) in myocardial I/R injury. Rats with I/R injuries had their myocardium directly injected with BMSCs-Exo. The outcomes demonstrated that cardiac function was enhanced and BMSCs-Exo dramatically decreased myocardial infarct size. Transcriptome sequencing was performed on heart tissues from the model and exosome-treated groups. GO and KEGG enrichment analyses revealed that exosomes might mitigate myocardial I/R damage via the AMPK/PGC-1α signaling pathway, confirmed by both in vitro and in vivo tests. The findings imply that compound C and sh-AMPK reverse the activation of PGC-1α and its downstream proteins and negate the protective effects of exosomes against oxidative stress and mitochondrial function in damaged cardiomyocytes. On the other hand, p-AMPK expression was unaffected by PGC-1α silencing. It was demonstrated that via activating the AMPK/PGC-1α signaling pathway, BMSCs-Exo might reduce oxidative stress and mitochondrial dysfunction in cardiomyocytes, thereby protecting against myocardial I/R damage.

Cardiovascular diseases (CVDs) represent a profound challenge with inflammation playing a significant role in their pathophysiology. Extracellular vesicles (EVs), which are membranous structures encapsulated by a lipid bilayer, are essential for intercellular communication by facilitating the transport of specific bioactive molecules, including microRNAs, proteins, and lipids. Emerging evidence suggests that the regulatory mechanisms governing cardiac resident cells are influenced by EVs, which function as messengers in intercellular communication and thereby contribute to the advancement of CVDs. In this review, we discuss the multifaceted biological functions of EVs and their involvement in the pathogenesis of various CVDs, encompassing myocardial infarction, ischemia-reperfusion injury, heart failure, atherosclerosis, myocarditis, cardiomyopathy, and aneurysm. Furthermore, we summarize the recent advancements in utilizing EVs as non-invasive biomarkers and in cell-free therapy based on EVs for the diagnosis and treatment of CVDs. Future research should investigate effective techniques for the isolation and purification of EVs from body fluids, while also exploring the pathways for the clinical translation of therapy based on EVs. Additionally, it is imperative to identify appropriate EV-miRNA profiles or combinations present in the circulation of patients, which could serve as biomarkers to improve the diagnostic accuracy of CVDs. By synthesizing and integrating recent research findings, this review aims to provide innovative perspectives for the pathogenesis of CVDs and potential therapeutic strategies.

Antibody-mediated rejection remains a leading cause of graft loss during kidney transplantation. Ischemia reperfusion injury (IRI) has been reported to promote T cell proliferation, leading to B cell activation and subsequent production of donor-specific antibodies, which target antigens on the vascular endothelium. We hypothesize that a novel therapeutic strategy targeting highly toxic reactive oxygen species could mitigate oxidative stress and immune responses associated with IRI. Our previous study demonstrated that oral administration of a silicon (Si)-based agent consistently generates substantial amounts of hydrogen, effectively suppressing IRI-induced oxidative stress and acute kidney injury in a rat renal clamp model. Here, we investigated the effect of the Si-based agent on immune responses in an allogeneic kidney transplant setting. Using both short-term and long-term evaluation models, we found that the Si-based agent suppressed oxidative stress and acquired immunity activation. Furthermore, early suppression of donor-specific antibody production and amelioration of chronic antibody-mediated rejection were observed. These findings indicate that the Si-based agent offers protective effects on graft function and survival, highlighting its potential clinical application to improve outcomes for kidney transplant recipients.

Stroke is the second-leading global cause of death. The damage attributed to the immune storm triggered by ischemia-reperfusion injury (IRI) post-stroke is substantial. However, data on the transcriptomic dynamics of pyroptosis in IRI are limited. This study aimed to analyze the expression of key pyroptosis genes in stroke and their correlation with immune infiltration. Pyroptosis-related genes were identified from the obtained middle cerebral artery occlusion (MCAO) datasets. Differential expression and functional analyses of pyroptosis-related genes were performed, and differences in functional enrichment between high-risk and low-risk groups were determined. An MCAO diagnostic model was constructed and validated using selected pyroptosis-related genes with differential expression. High- and low-risk MCAO groups were constructed for expression and immune cell correlation analysis with pyroptosis-related hub genes. A regulatory network between pyroptosis-related hub genes and miRNA was also constructed, and protein domains were predicted. The expression of key pyroptosis genes was validated using an MCAO rat model. Twenty-five pyroptosis genes showed differential expression, including four hub genes, namely WISP2, MELK, SDF2L1, and AURKB. Characteristic genes were verified using real-time quantitative PCR analyses. The high- and low-risk groups showed significant expression differences for WISP2, MELK, and SDF2L1. In immune infiltration analysis, 12 immune cells showed differences in expression in MCAO samples. Further analysis demonstrated significant positive correlations between the pyroptosis-related hub gene SDF2L1 and immune cell-activated dendritic cells in the high-risk group and immune cell natural killer cells in the low-risk group. This study identified four pyroptosis-related hub genes, with elevated WISP2, MELK, and SDF2L1 expression closely associated with the high-risk group. The analysis of inflammatory cell types in immune infiltration can predict ischemic stroke risk levels and help to facilitate treatment.

Intestinal ischemia-reperfusion (I/R) injury occurs under various surgical or disease conditions, where tissue hypoxia followed by reoxygenation results in the production of oxygen radicals and inflammatory mediators. These substances can target the endothelial barrier, leading to microvascular leakage. In this study, we induced intestinal I/R injury in mice by occluding the superior mesenteric artery, followed by removing the clamp to resume blood circulation. We assessed microvascular permeability to plasma proteins in vivo using intravital microscopy, measuring the time-dependent tracer distribution in the intravascular versus extravascular space in the mouse mesentery. Additionally, we examined endothelial cell-cell adhesive barrier resistance and junction morphology in cultured endothelial cell monolayers. At the molecular level, FAK inhibition similarly inhibited endothelial junction opening and barrier dysfunction in response to hydrogen peroxide-induced oxidative stress. To further investigate FAK's role with tissue/cell specificity, we developed an endothelial-specific inducible FAK knockout mouse model by crossbreeding FAK-floxed (FAKfl/fl) mice with Tie-2-CreERT2 transgenic mice. Compared to their wild-type controls, endothelial-specific FAK-deficient mice showed a blunted microvascular hyperpermeability response following I/R injury in the gut. Overall, our study demonstrates that FAK plays a significant signaling role in mediating endothelial barrier dysfunction and microvascular leakage during ischemia-reperfusion injury.

The outcome after liver transplantation has improved in recent years, which can be attributed to superior storage and transportation conditions of the organs, as well as better peri- and postoperative management and advancements in surgical techniques. Nevertheless, there is an increasing discrepancy between the need for organs and their availability. Consequently, the mortality rate on the waiting list is high and continues to rise. One way of counteracting this trend is to increase the use of "expanded criteria donors." This means that more and more donors will be included, especially those who are older and having additional comorbidities (eg, steatosis). A major complication of any transplantation is the occurrence of ischemia/reperfusion injury (IRI), which often leads to liver dysfunction and failure. However, there have been various promising approaches to minimize IRI in recent years, but an effective and clinically applicable method to achieve a better outcome for patients after liver transplantation is still missing. Thereby, the so-called marginal organs are predominantly affected by IRI; thus, it is crucial to develop suitable and effective treatment options for patients. Recently, regulated cell death mechanisms, particularly ferroptosis, have been implicated to play a major role in IRI, including the liver. Therefore, inhibiting this kind of cell death modality presents a promising therapeutic approach for the management of this yet untreatable condition. Thus, this review provides an overview of the role of ferroptosis in liver IRI and transplantation and discusses possible therapeutic solutions based on ferroptosis inhibition to restrain IRI in marginal organs (especially steatosis and donation after circulatory death organs).

To measure and validate elevated succinate in brain during circulatory arrest in a piglet model of cardiopulmonary bypass.

Using data from an archive of 3T 1H MR spectra acquired in previous in-magnet studies, dynamic plots of succinate, spectral simulations and difference spectra were generated for analysis and validation.

Elevation of succinate during circulatory arrest was observed and validated. Fitting bias was evaluated as a function of the line-widths and signal-to-noise ratios of the archived data. Succinate increases were independent of bypass temperature. Succinate elevation was also not observed with antegrade cerebral perfusion.

Although spectrally overlapped and at sub-millimolar levels, elevated brain succinate can be reliably measured by dynamic MR spectroscopy at 3T. Noise dependent bias of the stronger overlapping signals did not impact the succinate measurement. Elevated succinate during circulatory arrest and its recovery after reperfusion was observed. This finding is consistent with earlier reports that correlate elevated succinate with ischemic-reperfusion injury.

The superoxide anion radical (O2˙-) represents the primary reactive oxygen species generated in biological systems. Real-time monitoring of its dynamic fluctuations provides valuable insights into disease progression and enables early diagnosis of hepatic ischemia-reperfusion injury (HIRI). In this work, we developed a novel dual-color fluorescent carbon dot (CD) probe through a one-step hydrothermal synthesis for reversible O2˙- detection. The CDs demonstrated excellent sensitivity, dynamically detecting O2˙- concentrations ranging from 0 to 60 μM with a detection limit of 0.56 μM. The probe exhibited remarkable reversibility, maintaining stable performance through at least three complete oxidation-reduction cycles following glutathione (GSH) treatment. In practical applications, the CDs achieved 95.2-104% recovery rates when detecting O2˙- in serum samples. Cellular imaging experiments confirmed the probe's effectiveness in normal hepatocytes (LO2), showing clear reversible responses to O2˙- fluctuations. Application in a HIRI cell model revealed significant elevation of O2˙- levels and provided new evidence for its role in HIRI-related signaling pathways. This study not only presents an effective dual-color fluorescent probe for dynamic O2˙- monitoring but also establishes a versatile synthetic strategy that could be adapted for imaging other biologically relevant molecules in living cells.

The hypoxia-ischemia (H-I) diseases share some common mechanisms which may help to delay the diseases' processing. However, the shared features are still unclear due to the lack of large scale high-quality multi - omics data that specifically target the same disease, population, and tissues/cells. In this study, we developed a novel risk assessment method to analyze four H-I diseases including eclampsia/preeclampsia (PE), pulmonary arterial hypertension (PAH), high-altitude polycythemia (HAPC), and ischemic stroke (IS). A combined new evaluation score was designed to integrate evaluation information from genomics, transcriptomics, proteomics, and metabolomics in previous researches. Genes were then divided into different groups according to their risk assessment score. The most significant group (direct biomarkers) contained genes with direct evidence of association to H-I disease: PIEZO2 and HPGD (shared), TSIX and SAA1 (PAH - specific), GSTM1, DNTT, and IGKC (HAPC - specific), LEP, SERPINA3, and ARHGEF4 (PE - specific), CD3D, ITK, and RPL18A (IS - specific). The groups 'Intermediate crucial biomarkers' contained genes played important roles in H-I disease related biological processes: CXCL8 (shared), HBG2, GRIN2A, and FGFBP1 (PAH - specific), FAM111B (HAPC - specific), C12orf39 and SLAMF1 (PE - specific). The genes lacking disease-association evidence but with similar characteristics with the above two groups were considered as 'potential minor-effect biomarkers': are SRRM2 - AS1 (shared), ATP8A1 (PAH - specific), RXFP1 and HJURP (HAPC - specific), HIST1H1T (PE - specific). With the development of biological experiments, these intermediate crucial and potential minor-effect biomarkers may be proved to be direct biomarkers in the future. Therefore, these biomarkers may serve as an entry point for subsequent research and are of great significance.

Skeletal muscle injuries are short-term, that occur in people who play sports and train. Regular exercise and sports populations undergo repetitive tearing and regeneration of skeletal muscle, in which muscle damage is a necessary component to produce an oxidative inflammatory response and tissue reconstruction. The primary goals of treating this illness are to reduce the disease process cycle and get rid of symptoms like swelling and inflammation at the site of localized injury. Berberine (BBR) has several pharmacological effects, including anti-inflammatory, anti-tumor, and anti-arrhythmic properties.

In order to treat skeletal muscle injuries, a safe and non-toxic nanogel (BBR/GelMA) was developed for efficient berberine delivery. It also investigated whether BBR/GelMA had anti-inflammatory properties via the NF-κB pathway. Microwave irradiation was added to promote the uptake of BBR in BBR/GelMA by injured skeletal muscle and to accelerate the process of injury recovery.

It turns out that the survival rates of NIH313 and L929 cells decreased to varying degrees in GelMA loaded with different concentrations of BBR, but the survival rates of the two cell lines were the highest at a concentration of 0.125 mg/mL.

In this experiment, the inhibitory effect of BBR/GelMA on inflammation was studied. After NIH-313 and L929 cells were treated with GelMA loaded with different doses of BBR, it was found that the concentration of BBR/0.5 mg/mL had the best inhibitory effect on these two inflammation-inducing cell lines, and this inhibitory effect was related to the drug loading concentration. On the other hand, BBR/GelMA and microwave therapy can play an anti-inflammatory and repairing role in skeletal muscle through NF-κB pathway. In addition, microwave can accelerate the diffusion of BBR in BBR/GelMA within injured skeletal muscle, speeding up the healing process after skeletal muscle injury and shortening the disease cycle.

Preeclampsia is a hypertension condition of human pregnancy that poses a significant risk to pregnant women and their fetus. It complicates about 2-8% of human pregnancies worldwide and displays multifactorial pathogenesis, including increased placental oxidative stress because of disturbed utero-placental blood flow. Recent evidence suggests that increased oxidative stress promotes acceleration of the placental senescence which is implicated in the pathogenesis of preeclampsia. This review focuses on the mechanisms that lead to oxidative stress in preeclamptic patients and examines the role of oxidative stress-induced placental senescence in the pathogenesis of the disease.

Acupuncture is clinically effective in the treatment of ischemic stroke. The report suggests that energy stress inhibits ferroptosis in part through AMPK. However, whether the effect of activating AMPK is related to ferroptosis in ischemic stroke and whether acupuncture can achieve neuroprotection against ischemic brain injury through the pathway of inhibiting ferroptosis by activating AMPK has not been confirmed.

In this experiment, all rats were randomly divided into 4 groups: Sham group, MCAO/R group, MA (MCAO/R + acupuncture) group, and MAM (MCAO/R + acupuncture + metformin) group. The middle cerebral artery occlusion/reperfusion injury (MCAO/R) model was created by the wire embolism method, and the MA and MAM groups received acupuncture treatment with electrotherapy (1 mA, 2/15 Hz, 20 min each), while the MAM group continued to receive metformin (oral gavage 200 mg/kg) after successful modelling. Neurological deficit score and infarct volume were measured, Prussian blue staining and mitochondrial structural changes were observed, and Fe2+ and MDA levels were determined in the brain tissue of the rats. Western blot results were analyzed to determine differences in the expression of TFR1, SLC7A11, GPX4 and AMPK、p-AMPK proteins in order to explore the possible pathological processes involved in cerebral ischemia at behavioral, histological and molecular levels and the possible protective mechanisms of acupuncture.

Acupuncture attenuated ischemia-reperfusion-induced brain damage and mitochondrial damage. Further studies showed that acupuncture reduced the levels of Fe2+, MDA and the expression of TFR1 protein and increased the expression of SLC7A11 and GPX4 protein in the diseased hippocampal region of MCAO/R rats. In addition, metformin, as an AMPK activator, significantly enhanced the protective effect of acupuncture on cerebral ischemic injury and enhanced the acupuncture-mediated reduction of Fe2+, MDA levels and TFR1 protein expression and the increase of SLC7A11 and GPX4 protein expression in the lesioned hippocampal region of MCAO/R rats.

These findings suggest that acupuncture can inhibit ferroptosis and thus exert a protective effect against ischemic brain injury, and that this mechanism may be achieved by activating AMPK. This extends the mechanism of action of acupuncture in the treatment of ischemic stroke.

Remote ischemic preconditioning (RIPC) diminishes multi-organ failure induced by skeletal muscle ischemia and reperfusion (S-I/R). The current study investigated whether skeletal RIPC protection against S-I/R-induced acute lung injury (ALI) could be facilitated following simultaneous exposure to the glycoprotein hormone erythropoietin (EPO) in rats and whether this interaction is modulated by the NLRP3 inflammasome.

S-I/R challenge was performed by 3-h ischemia followed by 3-h reperfusion of the right hindlimb, whereas RIPC involved three 20-min brief consecutive I/R cycles of the contralateral hindlimb.

The lung injurious response to S-I/R was verified by: (i) decreases in minute respiratory volume (MRV), forced expiratory volume 1 (FEV1) and functional vital capacity (FVC), (ii) increases in respiratory rate (RR), (iii) falls in lung surfactant protein-D (SP-D) and rises in of lung plasminogen activator inhibitor-1 (PAI-1) and intercellular adhesion molecule-1 (ICAM-1), and (iv) disruption of alveolar architecture. These lung defects were partially amended by RIPC or EPO (500 or 5000 IU/kg). Further, the prior exposure to RIPC plus EPO-500 was more effective than separate interventions in rectifying ALI damages. Molecularly, the dual RIPC/EPO-500 regimen was also more effective in reversing the S-I/R-associated increments in pulmonary expressions of NLRP3 and related inflammatory (TLR4, MyD88, TRAF, NF-κB, TNF-α, IL-1β, and IL-18), apoptotic (ASC, procaspse-1, caspase-1), and microRNA signals (increases in miR-21 and decreases miR-495).

These findings suggest a pivotal role for the suppression of NLRP3 inflammasome and interconnected cellular offenses in the augmented therapeutic potential of the RIPC/EPO-500 regimen against S-I/R-induced ALI.

Solid organ transplantation is hampered by complications that arise after ischemia-reperfusion injury (IRI), a detrimental type of injury for which no adequate treatment options are available. Ferroptosis, an iron-dependent form of regulated cell death, is a major driver of IRI. This systematic review and meta-analysis summarizes the effects of pharmacological ferroptosis inhibition in abdominal organs in the setting of IRI. PubMed, Embase, Web of Science and Cochrane were searched for concepts "ferroptosis" and "IRI" in August 2023. To allow for meta-analyses, inhibitors were divided into different intervention pathways: (I) lipophilic radical scavengers, (II) iron chelators, (III) antioxidants, (IV) lipid metabolism inhibitors, (V) combination treatments, and (VI) others. When available, organ function and injury effect sizes were extracted and used for random-effects meta-analyses. In total 79 articles were included, describing 59 unique inhibitors in kidney, liver, and intestinal IRI. No studies in pancreas were found. Overall bias and study quality was unclear and average to low, respectively. Apart from 1 clinical study, all inhibitors were tested in preclinical settings. The vast majority of the studies showed ferroptosis inhibition to be protective against IRI under various treatment conditions. In liver and kidney IRI, meta-analyses on standardized effect sizes from 43 articles showed a combined protective effect against IRI compared with a nontreated controls for all analyzed intervention pathways. In conclusion, ferroptosis inhibition protects against abdominal IRI in preclinical research. Important questions regarding optimal intervention pathway, bioavailability, optimal dosage, side effects etc. should be addressed before clinical introduction.

This Part 1 of a bipartite review commences with a succinct exposition of innate alloimmunity in light of the danger/injury hypothesis in Immunology. The model posits that an alloimmune response, along with the presentation of alloantigens, is driven by DAMPs released from various forms of regulated cell death (RCD) induced by any severe injury to the donor or the donor organ, respectively. To provide a strong foundation for this review, which examines RCD and DAMPs as biomarkers and therapeutic targets in normothermic regional perfusion (NRP) and normothermic machine perfusion (NMP) to improve outcomes in organ transplantation, key insights are presented on the nature, classification, and functions of DAMPs, as well as the signaling mechanisms of RCD pathways, including ferroptosis, necroptosis, pyroptosis, and NETosis. Subsequently, a comprehensive discussion is provided on major periods of injuries to the donor or donor organs that are associated with the induction of RCD and DAMPs and precede the onset of the innate alloimmune response in recipients. These periods of injury to donor organs include conditions associated with donation after brain death (DBD) and donation after circulatory death (DCD). Particular emphasis in this discussion is placed on the different origins of RCD-associated DAMPs in DBD and DCD and the different routes they use within the circulatory system to reach potential allografts. The review ends by addressing another particularly critical period of injury to donor organs: their postischemic reperfusion following implantation into the recipient-a decisive factor in determining transplantation outcome. Here, the discussion focuses on mechanisms of ischemia-induced oxidative injury that causes RCD and generates DAMPs, which initiate a robust innate alloimmune response.

Custodiol is a well-established preservation solution for organ transplantation and was the basis for the development of Custodiol-N to improve graft preservation. Previous results in coronary artery bypass graft surgery have shown effective cardiac protection without safety concerns. This study aimed to evaluate the safety and ability of Custodiol-N to preserve cardiac grafts for heart transplantation.

This prospective, randomized, single-blind, multicenter, noninferiority study was conducted at three centers in Austria and Germany. The primary end-point was creatine kinase (CK-MB) peak value from 4 to 168 hours after opening of the aortic cross clamp, with a 30% noninferiority margin. Key secondary efficacy end-points include patient and graft survival, incidence of primary graft failure, or length of stay in the intensive care unit. The primary and secondary end-points were analyzed in both the treated and per protocol populations.

A total of 105 randomized patients received Custodiol (n = 52) or Custodiol-N (n = 53) preserved hearts. Average donor age and ischemic times were comparable. Average CK-MB peak values were 176.94 ± 189.61 U/L for the Custodiol versus 130.51 ± 69.60 U/liter for the Custodiol-N group (p-value for noninferiority of Custodiol-N by 30% <0.0001). Patient survival was comparable 1-year post transplantation (90.4% for Custodiol versus 88.7% for Custodiol-N). The incidence of primary graft failure and median length of intensive care unit stay were higher for Custodiol group. Safety assessment showed evenly distributed adverse events.

This study shows that Custodiol-N is safe, noninferior, and provides similar cardiac graft protection as the established Custodiol solution.

The pathway of testicular ischemia-reperfusion injury (TIRI) has been shown to involve reactive oxygen species (ROS) generation in the ischemic phase and later phase of reperfusion. This study was therefore designed to investigate the effect of blockage of ROS in the ischemic and reperfusion phases of TIRI. Thirty male Wistar rats were grouped into five groups (n = 6 rats each): sham, torsion + detorsion (TD), febuxostat (FEB)-administered (TFD) group, vitamin E (V)-administered (TDV) group, and FEB and vitamin E-administered (TFDV) group. Blood samples (for inflammatory and hormonal assay), testicular (for oxidative stress and histopathology), and epididymal (for sperm DNA damage and indices) tissues were collected after 3 days of detorsion. The TFD and TFDV groups showed a significant reduction in XO and MDA (p < 0.001; η2 > 0.7), as well as a concomitant increase in CAT, thiols, and SOD levels when compared with the TD group (p < 0.01, η2 > 0.5). The TFD group significantly reduced all inflammatory markers (p < 0.05; η2 = 0.75). The observed increase (p < 0.05; η2 = 0.92) in LH level, in response to a low level of testosterone in the TD group, was significantly raised in TFD and TFDV groups. The observed decrease (p < 0.001) in inhibin level in the TD group was raised (p < 0.05; η2 = 0.90) in the TDV group only. A significant increase (p < 0.001) in sperm DNA damage in the TD group was significantly reduced (p < 0.05; η2 = 0.88) in all the treatment groups while the reduced sperm viability (p < 0.01) in the TD group was increased (p < 0.05) in the TFDV group only. There was an improvement in the testicular cytoarchitecture in the TFD and TFDV groups. This study showed that sequential administration of febuxostat in the ischemic phase of TT and vitamin E in the later phase of reperfusion protects the testes against TIRI via inhibition of oxidative stress, inflammation, and sperm DNA damage.

Renal ischemia-reperfusion injury (IRI) is a common complication in several clinical scenarios including kidney transplantation. Mannan-binding lectin-associated serine proteinase (MASP)-2 is essential for activation of the complement lectin pathway, which has been implicated in the pathogenesis of renal IRI and therefore represents a potential therapeutic target. We developed a new, affinity-enhanced MASP-2 inhibitor, EVO24, by directed evolution of the D2 domain of human tissue factor pathway inhibitor. EVO24 was fused with a human IgG1-Fc to create the homodimer EVO24L, which potently and selectively inhibited the lectin pathway in human and mouse serum in vitro. EVO24L was tested in a mouse model of unilateral warm renal IRI. EVO24L administered before and after ischemia significantly protected against IRI, with improved renal function as well as reduced tubular injury and inflammatory cell infiltration at 24 h compared to vehicle-treated mice. Immunofluorescence analyses showed reduced deposition of complement components (C3d, C4d, and C9) and reduced expression of VCAM-1, indicating a decrease in complement activation and endothelial cell activation. Additionally, EVO24L treatment lowered plasma levels of complement C5a, hyaluronan (a marker of endothelial glycocalyx shedding), and the proinflammatory cytokines IL-6 and TNF-α. Our findings indicate that EVO24L inhibits acute inflammatory responses in renal IRI by blocking the lectin pathway, confirming the important role of this pathway in acute ischemic kidney injury and warranting further investigation of EVO24L in clinical settings.

Cerebral ischaemia-reperfusion injury (CIRI) is a major contributor to global morbidity and mortality, although its underlying mechanisms remain only partly understood. Emerging evidence indicates that inhibiting microglia-mediated neuroinflammation would be an effective therapeutic approach for CIRI, and pharmacological interventions targeting this pathway hold significant therapeutic promise. This study aimed to identify a potent anti-inflammatory drug from a natural compound library as a potential treatment for CIRI.

We used oxygen-glucose deprivation/reperfusion (OGD/R) and middle cerebral artery occlusion in male C57BL/6 mice to evaluate the efficacy of DHA in neurological deficits and the anti-inflammatory effects. Using BV2 cells and murine brain tissue, liquid chromatography-tandem mass spectrometry was used to identify potential molecular targets of DHA, followed by bio-layer interferometry, molecular docking, molecular dynamics simulations and cellular thermal shift assays to validate DHA's binding interactions with protein kinase C delta (PKCδ).

DHA decreased production of pro-inflammatory cytokines following OGD/R, thereby inhibiting microglia-mediated neuroinflammation to protect neurons and reducing brain infarct size and improving neurological outcomes. Mechanistically, DHA directly bound to PKCδ, inhibiting its phosphorylation and downstream NF-κB signalling. This binding interaction involved TRP55 and LEU106 on PKCδ, as confirmed by molecular docking and other biophysical techniques.

DHA specifically interacts with PKCδ, preventing its phosphorylation induced by ischaemia-reperfusion injury. These results suggest that DHA is a novel inhibitor of PKCδ and provide solid experimental foundations for using DHA in treating neuroinflammation-related conditions, such as CIRI.

Caloric restriction mimetics (CRMs) replicate the positive effects of caloric restriction (CR) and have demonstrated therapeutic benefits in neuroinflammatory and cardiovascular diseases. However, it remains uncertain whether CRMs enhance functional recovery following ischemia/reperfusion (I/R) injury, as well as the specific mechanisms involved in this process. This study examines the therapeutic potential of the CRM 3,4-dimethoxychalcone (3,4-DC) in limb I/R injury. Histology, tissue swelling analysis, and laser doppler imaging (LDI) were used to assess the viability of the limbs. Western blotting and immunofluorescence were utilized to examine apoptosis levels, oxidative stress (OS), autophagy, transcription factor EB (TFEB) activity, and mucolipin 1 (MCOLN1)-calcineurin signaling pathway. The administration of 3,4-DC notably alleviated hypoperfusion, tissue swelling, skeletal muscle fiber damage, and cellular injury in the limb caused by I/R. The pharmacological blockade of autophagy negated the antioxidant and antiapoptotic effects of 3,4-DC. Moreover, RNA interference-mediated TFEB silencing eliminated the 3,4-DC-induced restoration of autophagy, antioxidant response, and antiapoptotic effects. Additionally, our findings revealed that 3,4-DC modulates TFEB activity via the MCOLN1-calcineurin signaling pathway. 3,4-DC facilitates functional recovery by enhancing TFEB-driven autophagy, while simultaneously suppressing oxidative stress and apoptosis following I/R injury, suggesting its potential value in clinical applications.