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Background/Objectives: Ischemic stroke is a leading cause of disability and mortality worldwide, with current therapies limited in addressing its complex pathophysiological mechanisms, such as inflammation, oxidative stress, apoptosis, and impaired autophagy. Glycyrrhizic acid (GA), a bioactive compound from licorice (Glycyrrhiza glabra L.), has demonstrated neuroprotective properties in preclinical studies. This review consolidates current evidence on GA’s pharmacological mechanisms and assesses its potential as a therapeutic agent for ischemic stroke. Methods: This review examines findings from recent preclinical studies and reviews on GA’s neuroprotective effects, focusing on its modulation of inflammation, oxidative stress, apoptosis, and autophagy. Studies were identified from major scientific databases, including PubMed, Web of Science, and Embase, covering research from January 2000 to August 2024. Results: GA has demonstrated significant neuroprotective effects through the modulation of key pathways, including HMGB1/TLR4/NF-κB and Keap1/Nrf2, thereby reducing neuroinflammation, oxidative stress, and apoptosis. Additionally, GA promotes autophagy and modulates immune responses, suggesting it could serve as an adjunct therapy to enhance the efficacy and safety of existing treatments, such as thrombolysis. Conclusions: Current findings underscore GA’s potential as a multi-targeted neuroprotective agent in ischemic stroke, highlighting its anti-inflammatory, antioxidant, and anti-apoptotic properties. However, while preclinical data are promising, further clinical trials are necessary to validate GA’s therapeutic potential in humans. This review provides a comprehensive overview of GA’s mechanisms of action, proposing directions for future research to explore its role in ischemic stroke management.

This study aimed to clarify the roles and underlying mechanisms of luteolin in the progression of cerebral ischemia/reperfusion injury (CIRI).

BACKGROUND: Croton cream can activate ERK pathways and have anti-apoptotic effects on neuronal cells. It is not clear whether it synergistically exerts antiapoptotic effects by inhibiting the activation of JNK and p38 pathways. OBJECTIVE: To explore the effects and mechanisms of croton cream on neuronal damage and apoptosis in the ischemic cortex of rats with cerebral ischemiareperfusion injury. METHODS: (1) Ninety Sprague-Dawley rats were randomly divided into sham operation group, model group, croton cream low-dose group, croton cream medium-dose group, croton cream high-dose group and nimodipine group, with 15 rats in each group. Except for the sham operation group, animal models of middle cerebral artery occlusion were prepared in rats by the thread method. Rats in the three croton cream groups were given 20, 40, and 60 mg/kg croton cream, respectively. Rats in the sham operation and model groups were given the same amount of normal saline, once a day, for 7 consecutive days. The optimal concentration of croton cream, namely the high dose of croton cream, was selected based on neurological deficit score, TTC staining, brain tissue water content, hematoxylin-eosin staining and Nissl staining. (2) Another 120 Sprague-Dawley rats were randomly divided into sham operation group, model group, croton cream group, JNK inhibitor group, croton cream+JNK inhibitor group, p38 MAPK inhibitor group, croton cream+p38 MAPK inhibitor group, and nimodipine group, with 15 rats in each group. Animal models of middle cerebral artery occlusion were prepared using the thread method in all the groups except in the sham operation group. Thirty minutes before modeling, 10 μL of SP600125 (JNK inhibitor) and 10 μL of SB203580 (p38 MAPK inhibitor) were injected into the lateral ventricle of the rats, respectively. Rats in croton cream groups were intragastrically given 60 mg/kg croton cream. Seven days later, the JNK/p38 MAPK signaling pathway, apoptosis-related proteins and cell apoptosis were detected by western blot, TUNEL staining and flow cytometry, respectively. RESULTS AND CONCLUSION: (1) Compared with the sham operation group, neurological deficit score, cerebral water content, cerebral infarction volume and apoptosis rate were significantly increased in the model group (P < 0.05), where nerve cells showed scattered distribution. Compared with the model group, neurological deficit score, water content of brain tissue and cerebral infarction volume were significantly decreased in the croton cream medium-dose group, high-dose group and nimodipine group (P < 0.05), and the pathological morphology of nerve cells was significantly improved. (2) Compared with the JNK inhibitor group, p-JNK/JNK, p-p38/p38 and Bax expressions in rat brain tissue and the apoptotic rate were significantly decreased in the croton cream+inhibitor groups (P < 0.05), while the expression of and Bcl-2 was significantly increased (P < 0.05). To conclude, croton cream may inhibit the activation of JNK/p38 MAPK signaling pathway and reduce neuronal apoptosis to achieve neuroprotective effects in rats with cerebral ischemia-reperfusion injury. [ABSTRACT FROM AUTHOR]

Objective: To investigate the role of HMGB1-TLR4-NF-κB signaling pathway in cerebral ischemia-reperfusion in-jury and the effect of adenosine preconditioning on the signaling pathway. Methods: Total 80 adult male Sprague-Dawley rats weighing 220~270 g were selected from the Animal Center of Xinxiang Medical University. The rats were randomly divided into F group (sham operation group, I/R group (ischemia reperfusion group) and AP group (adenosine preconditioning group). The MCAO model of rats was established by wire embolization. Quantitative analysis of neural function in successfully modeled rats using animal behavior scoring method, the morphological changes of brain cells were observed by HE staining, TTC staining was used to observe cerebral infarction and cerebral infarction volume was calculated; Immunohistochemical staining was used to detect HMGB1, TLR4 and NF-κB protein expression levels in brain tissues of each group. The data were statistically analyzed by one-way ANOVA in SPSS26.0 software. Results: After ischemia reperfusion, the neurological function of I/R group and AP group showed different degrees of impairment, and the neurological function scores of the two groups were significantly higher than that of F group, the difference was statistically significant (P<0.05 ), and the neurological function of the AP group was significantly less than that of I/R group, the difference was also statistically significant (P<0.05). TTC staining showed that AP group, I/R group rat cerebral infarction volume was significantly more than F group [ (93.670±4.509) mm³, (123.670±7.234) mm³ vs (0.000±0.000) mm³], and AP group rats infarction volume was significantly reduced than that in I/R group, the difference had statistical significance (P<0.05). Immunohistochemistry showed that HMGB1, TLR4, NF-κB protein in F group with a small amount of expressions in rats, while significantly expressed in AP group and I/R group relatively, and the AP group of each subgroup rat HMGB1, TLR4, NF-κB protein expressions significantly lower than the amount of I/R group, the difference had statistical significance (P<0.05). Conclusion: Adenosine preconditioning can reduce the expressions of HMGB1, TLR4 and NF-κB protein, and then protect the rats with cerebral ischemia-reperfusion injury. [ABSTRACT FROM AUTHOR]

Cerebral ischemia-reperfusion (I/R) injury is the predominant causes for the poor prognosis of ischemic stroke patients after reperfusion therapy. Currently, potent therapeutic interventions for cerebral I/R injury are still very limited. Melatonin, an endogenous hormone, was found to be valid in preventing I/R injury in a variety of organs. However, a systematic review covering all neuroprotective effects of melatonin in cerebral I/R injury has not been reported yet. Thus, we perform a comprehensive overview of the influence of melatonin on cerebral I/R injury by collecting all available literature exploring the latent effect of melatonin on cerebral I/R injury as well as ischemic stroke. In this systematic review, we outline the extensive scientific studies and summarize the beneficial functions of melatonin, including reducing infarct volume, decreasing brain edema, improving neurological functions and attenuating blood-brain barrier breakdown, as well as its key protective mechanisms on almost every aspect of cerebral I/R injury, including inhibiting oxidative stress, neuroinflammation, apoptosis, excessive autophagy, glutamate excitotoxicity and mitochondrial dysfunction. Subsequently, we also review the predictive and therapeutic implications of melatonin on ischemic stroke reported in clinical studies. We hope that our systematic review can provide the most comprehensive introduction of current advancements on melatonin in cerebral I/R injury and new insights into personalized diagnosis and treatment of ischemic stroke. [ABSTRACT FROM AUTHOR]

Background: This study aimed to investigate dynamic urinary proteome changes of electroacupuncture (EP) on cerebral ischemia‑reperfusion (CI/R) injured rats and to explore the therapeutic biological mechanisms of EP. Methods: First, changed urinary proteins were found in EP stimulation in healthy rats. Then, we used a CI/R injury rat model induced by Pulsinelli's four‐vessel occlusion (4‐VO) method to explore the function of EP on urinary proteome in CI/R injury. Urine samples were collected for proteome analysis by liquid chromatography‐tandem mass spectrometry (LC‐MS/MS) and bioinformatics analysis. Results: In total, 384 proteins were identified, among which 47 proteins (23 upregulated, 24 downregulated) were differentially expressed with 0.6‐log FC and p <.05. Gene ontology analysis revealed that the cell redox homeostasis, acute‐phase response, response to lipopolysaccharide, and cellular response to glucocorticoid stimulus were significantly enriched. The partially biologically connected differential proteins were found by the Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis in the EP group. With the CI/R rat model, 80 proteins (27 upregulated, 53 downregulated) were significantly changed in the CI/R rats compared to the controls. Among these differentially expressed proteins (DEPs), 23 proteins (17 upregulated, six downregulated) showed significant changes after EP treatment (0.6‐log FC change, p <.05). The main related biological processes were aging, immune response, acute‐phase response, liver regeneration, protein catabolic process, and response to oxidative stress. Many metabolic pathways were enriched by KEGG analysis. Conclusion: Our results indicate that the EP could alleviate cerebral damage induced by ischemia‑reperfusion through an anti‐inflammatory and metabolism regulation mechanism. The urinary proteome might reflect the pathophysiological changes in EP pretreatment in the treatment and prevention of CI/R injury. [ABSTRACT FROM AUTHOR]

Background: Stroke is a debilitating condition with high morbidity, disability, and mortality that significantly affects the quality of life of patients. In China, the WenYang FuYuan recipe is widely used to treat ischemic stroke. However, the underlying mechanism remains unknown, so exploring the potential mechanism of action of this formula is of great practical significance for stroke treatment. Objective: This study employed network pharmacology, molecular docking, and in vivo experiments to clarify the active ingredients, potential targets, and molecular mechanisms of the WenYang FuYuan recipe in cerebral ischemia-reperfusion injury, with a view to providing a solid scientific foundation for the subsequent study of this recipe. Materials and methods: Active ingredients of the WenYang FuYuan recipe were screened using the traditional Chinese medicine systems pharmacology database and analysis platform. Network pharmacology approaches were used to explore the potential targets and mechanisms of action of the WenYang FuYuan recipe for the treatment of cerebral ischemia-reperfusion injury. The Middle Cerebral Artery Occlusion/Reperfusion 2 h Sprague Dawley rat model was prepared, and TTC staining and modified neurological severity score were applied to examine the neurological deficits in rats. HE staining and Nissl staining were applied to examine the pathological changes in rats. Immunofluorescence labeling and Elisa assay were applied to examine the expression levels of certain proteins and associated factors, while qRT-PCR and Western blotting were applied to examine the expression levels of linked proteins and mRNAs in disease-related signaling pathways. Results: We identified 62 key active ingredients in the WenYang FuYuan recipe, with 222 highly significant I/R targets, forming 138 pairs of medication components and component-targets, with the top five being Quercetin, Kaempferol, Luteolin, β-sitosterol, and Stigmasterol. The key targets included TP53, RELA, TNF, STAT1, and MAPK14 (p38MAPK). Targets related to cerebral ischemia-reperfusion injury were enriched in chemical responses, enzyme binding, endomembrane system, while enriched pathways included lipid and atherosclerosis, fluid shear stress and atherosclerosis, AGE-RAGE signaling in diabetic complications. In addition, the main five active ingredients and targets in the WenYang FuYuan recipe showed high binding affinity (e.g. Stigmasterol and MAPK14, total energy

Objective: To explore the preventive effect of electroacupuncture pretreatment on stroke in rats by inhibiting ferroptosis and oxidative stress. Methods: Rats were randomly assigned to the sham, middle cerebral artery occlusion/reperfusion (MCAO/R), MCAO/R + EP, MCAO/R + EP + erastin, and MCAO/R + EP + ferrostatin 1 groups. Daily electroacupuncture was performed 2 weeks before establishing the MCAO/R model utilizing the modified Zea Longa suture method. Rats were sacrificed 1 day after reperfusion, and brain tissues were collected. They were prepared for hematoxylin and eosin staining, prussian blue staining, transmission electron microscope. Measurement of total iron levels using a commercial kit, detection of malondialdehyde (MDA) and superoxide dismutase (SOD) levels by ELISA, and examination of 15-lox2, GPX4, SLC7A11, ACSL4, and TFR1 by western blotting. Results: Compared with sham rats, cerebral infarction size was dramatically larger in MCAO/R rats. Moreover, the MCAO/R group displayed damaged mitochondria with a disarranged structure of cristae; free iron, total iron levels, and oxidative stress were significantly higher. Cerebral pathological lesions, oxidative stress, total iron levels, and protein levels of ACSL4, TFR1, and 15-lox2 were significantly reduced in the MCAO/R + EP and MCAO/R + EP + ferrostatin 1 groups, while the protective effect of electroacupuncture pretreatment on cerebral ischemia-reperfusion injury was inhibited by treatment with the ferroptosis activator erastin. Conclusion: Electroacupuncture pretreatment can protect rats from cerebral ischemia-reperfusion injury by reducing the area of cerebral infarction and inhibiting ferroptosis and oxidative stress.

Objective: This article aims to identify genetic features associated with immune cell infiltration in cerebral ischemia-reperfusion injury (CIRI) development through bioinformatics, with the goal of discovering diagnostic biomarkers and potential therapeutic targets. Methods: We obtained two datasets from the Gene Expression Omnibus (GEO) database to identify immune-related differentially expressed genes (IRDEGs). These genes' functions were analyzed via Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG). Tools such as CIBERSORT and ssGSEA assessed immune cell infiltration. The Starbase and miRDB databases predicted miRNAs interacting with hub genes, and Cytoscape software mapped mRNA-miRNA interaction networks. The ENCORI database was employed to predict RNA binding proteins interacting with hub genes. Key genes were identified using a random forest algorithm and constructing a Support Vector Machine (SVM) model. LASSO regression analysis constructed a diagnostic model for hub genes to determine their diagnostic value, and PCR analysis validated their expression in cerebral ischemia-reperfusion. Results: We identified 10 IRDEGs (C1qa, Ccl4, Cd74, Cd8a, Cxcl10, Gmfg, Grp, Lgals3bp, Timp1, Vim). The random forest algorithm, and SVM model intersection revealed three key genes (Ccl4, Gmfg, C1qa) as diagnostic biomarkers for CIRI. LASSO regression analysis, further refined this to two key genes (Ccl4 and C1qa), With ROC curve, analysis confirming their diagnostic efficacy (C1qa AUC = 0.75, Ccl4 AUC = 0.939). PCR analysis corroborated these findings. Conclusions: Our study elucidates immune and metabolic response mechanisms in CIRI, identifying two immune-related genes as key biomarkers and potential therapeutic targets in response to cerebral ischemia-reperfusion injury.

Abstract This study was to probe the protective effects and mechanisms of salvianolic acid A (SAA) on cerebral ischemia–reperfusion injury (CIRI). The middle cerebral artery occlusion model (MCAO) was established in rats. Rats' behavior, neurological deficits, brain injury, inflammation, and apoptosis in the brain tissue were evaluated. The inflammatory response and apoptosis of PC12 cells induced by oxygen glucose deprivation/reperfusion (OGD/R) were detected. SAA‐mediated changes in miR‐212‐3p, SOX7, and Wnt/β‐catenin pathway were determined, and the targeting relationship between miR‐212‐3p and SOX7 was clarified. SAA alleviated the neurological deficits and brain injury of MCAO rats and inhibited the inflammatory response and apoptosis of OGD/R‐conditioned PC‐12 cells. SAA upregulated miR‐212‐3p, Wnt3a, and β‐catenin, whereas inhibited SOX7 levels. Silencing miR‐212‐3p counteracted the protective effect of SAA in the context of CIRI. SOX7 was a target protein of miR‐212‐3p. Silencing SOX7 based on SAA and miR‐212‐3p knockdown suppressed OGD/R‐induced inflammation and apoptosis and increased Wnt3a and β‐catenin levels in PC12 cells. SAA can improve the brain and nervous system injury caused by cerebral ischemia–reperfusion by upregulating miR‐212‐3p, thereby inhibiting SOX7 and activating the Wnt/βcatenin signaling pathway.

Abstract Background and purpose Cerebral ischemia‒reperfusion injury causes significant harm to human health and is a major contributor to stroke-related deaths worldwide. Current treatments are limited, and new, more effective prevention and treatment strategies that target multiple cell components are urgently needed. Leucine-rich alpha-2 glycoprotein 1 (Lrg1) appears to be associated with the progression of cerebral ischemia‒reperfusion injury, but the exact mechanism of it is unknown. Methods Wild-type (WT) and Lrg1 knockout (Lrg1 −/−) mice were used to investigate the role of Lrg1 after cerebral ischemia‒reperfusion injury. The effects of Lrg1 knockout on brain infarct volume, blood‒brain barrier permeability, and neurological score (based on 2,3,5-triphenyl tetrazolium chloride, evans blue dye, hematoxylin, and eosin staining) were assessed. Single-cell RNA sequencing (scRNA-seq), immunofluorescence, and microvascular albumin leakage tests were utilized to investigate alterations in various cell components in brain tissue after Lrg1 knockout. Results Lrg1 expression was increased in various cell types of brain tissue after cerebral ischemia‒reperfusion injury. Lrg1 knockout reduced cerebral edema and infarct size and improved neurological function after cerebral ischemia‒reperfusion injury. Single-cell RNA sequencing analysis of WT and Lrg1 −/− mouse brain tissues after cerebral ischemia‒reperfusion injury revealed that Lrg1 knockout enhances blood‒brain barrier (BBB) by upregulating claudin 11, integrin β5, protocadherin 9, and annexin A2. Lrg1 knockout also promoted an anti-inflammatory and tissue-repairing phenotype in microglia and macrophages while reducing neuron and oligodendrocyte cell death. Conclusions Our results has shown that Lrg1 mediates numerous pathological processes involved in cerebral ischemia‒reperfusion injury by altering the functional states of various cell types, thereby rendering it a promising therapeutic target for cerebral ischemia‒reperfusion injury.

Cerebral ischemia-reperfusion injury (IRI), occurring after blood supply restoration, contributes significantly to stroke-related deaths. This study explored the combined impact and mechanisms of astragaloside IV (AS-IV), hydroxysafflor yellow A (HSYA), and their combination in mitigating IRI. Male Sprague–Dawley (SD) rats were randomized to the Sham, MCAO, MCAO+AS-IV, MCAO+HSYA, and MCAO+AS-IV+HSYA groups. Neurological deficits and cerebral infarction were examined after restoring the blood supply to the brain. Pathomorphological changes in the cerebral cortex were observed via HE staining. IL-1β and IL-18 were quantified using ELISA. The expression of NF-κB and GSDMD in the ischemic cerebrum was analyzed using immunohistochemistry. The expression levels of NLRP3, ASC, IL-1β, Caspase-1, and GSDMD in the ischemic cerebrum were evaluated using Western blot. The MCAO+AS-IV, MCAO+HSYA, and MCAO+AS-IV+HSYA groups exhibited notably better neurological function and cerebral infarction compared with the MCAO group. The combined treatment demonstrated superior brain tissue injury alleviation. Reductions in NF-κB, GSDMD positive cells, and NLRP3/ASC/IL-1β/Caspase-1/GSDMD protein expression in the ischemic brain were significantly more pronounced with the combined therapy, indicating a synergistic effect in countering cerebral IRI via the NF-κB/NLRP3/Caspase-1/GSDMD pathway inhibition of cell pyroptosis-induced injury.

Hemorrhagic transformation (HT) and restrictive therapeutic time window are significant limitations of delayed t-PA (tissue-type plasminogen activator) thrombolytic therapy in ischemic stroke patients. Plant Momordica charantia-derived exosome-like nanovesicles (MC-ELNs) can protect the blood–brain barrier (BBB) and inhibit neuronal apoptosis in stroke rats by inhibiting matrix metalloproteinase 9(MMP-9) expression. This study explored the therapeutic function and underlying mechanisms of MC-ELNs in treating HT. In delayed t-PA-treated ischemia-reperfusion rats, MC-ELNs significantly reduced mortality, HT, and cell apoptosis; MC-ELNs improved neurological function and BBB's integrity. MC-ELNs reduced ONOO− and MMP-9 expression and inhibited the release of HMGB1 from intracellular to extracellular in vivo and in vitro. ONOO− donor SIN-1 directly induced extracellular secretion of HMGB1, MC-ELNs and FeTmPyP (ONOO− decomposition catalyst, PDC) significantly reduced the expression of HMGB1 and inhibited MMP-9 activation in vitro. These findings indicate that MC-ELNs could protect BBB integrity and improve t-PA-induced HT by inhibiting the ONOO−/HMGB1/MMP-9 pathway.

Background: Xingnaojing (XNJ) injection, an extract derived from traditional Chinese medicine, is commonly used to treat ischemic stroke (IS). Previous studies have shown that XNJ has the ability to alleviate apoptosis in cerebral ischemia-reperfusion injury. However, the potential mechanisms have not been clarified. Objective: To identify the neuroprotective effect of XNJ and explore whether XNJ inhibits cell apoptosis associated with endoplasmic reticulum stress (ERS) after IS. Methods: In this study, cultured hippocampal neurons from mouse embryos and Sprague-Dawley rats were assigned randomly to four groups: sham, model, XNJ, and edaravone. The treatment groups were administered 2 h after modelling. Neurological deficit scores and motor performance tests were performed after 24 h of modelling. Additionally, pathomorphology, cell apoptosis and calcium content were evaluated. To ascertain the expression of ERS proteins, western blotting and polymerase chain reaction were employed. Results: The results indicated that XNJ treatment resulted in a notable decrease in infarct volume, apoptosis and missteps compared with the model group. XNJ also exhibited improvements in neurological function, grip strength and motor time. The calcium content significantly reduced in XNJ group. The XNJ administration resulted in a reduction in the levels of proteins associated with ERS including CHOP, GRP78, Bax, caspase-12, caspase-9, and cleaved-caspase-3, but an increase of the Bcl-2/Bax ratio. Furthermore, the downregulation of mRNA expression of CHOP, GRP78, caspase-12, caspase-9, and caspase-3 was confirmed in both cultured neurons and rat model. Conclusion: These findings suggest that XNJ may alleviate apoptosis by modulating the ERS-induced apoptosis pathway, making it a potential novel therapeutic approach for ischemic stroke.

Abstract Background This study aimed to investigate dynamic urinary proteome changes of electroacupuncture (EP) on cerebral ischemia‑reperfusion (CI/R) injured rats and to explore the therapeutic biological mechanisms of EP. Methods First, changed urinary proteins were found in EP stimulation in healthy rats. Then, we used a CI/R injury rat model induced by Pulsinelli's four‐vessel occlusion (4‐VO) method to explore the function of EP on urinary proteome in CI/R injury. Urine samples were collected for proteome analysis by liquid chromatography‐tandem mass spectrometry (LC‐MS/MS) and bioinformatics analysis. Results In total, 384 proteins were identified, among which 47 proteins (23 upregulated, 24 downregulated) were differentially expressed with 0.6‐log FC and p

Background Cerebral ischemia (CI), ranking as the second leading global cause of death, is frequently treated by reestablishing blood flow and oxygenation. Paradoxically, this reperfusion can intensify tissue damage, leading to CI-reperfusion injury. This research sought to uncover biomarkers pertaining to protein processing in the endoplasmic reticulum (PER) during CI-reperfusion injury. Methods We utilized the Gene Expression Omnibus (GEO) dataset GSE163614 to discern differentially expressed genes (DEGs) and single out PER-related DEGs. The functions and pathways of these PER-related DEGs were identified via Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. Core genes were pinpointed through protein-protein interaction (PPI) networks. Subsequent to this, genes with diagnostic relevance were distinguished using external validation datasets. A single-sample gene-set enrichment analysis (ssGSEA) was undertaken to pinpoint genes with strong associations to hypoxia and apoptosis, suggesting their potential roles as primary inducers of apoptosis in hypoxic conditions during ischemia-reperfusion injuries. Results Our study demonstrated that PER-related genes, specifically ADCY5, CAMK2A, PLCB1, NTRK2, and DLG4, were markedly down-regulated in models, exhibiting a robust association with hypoxia and apoptosis. Conclusion The data indicates that ADCY5, CAMK2A, PLCB1, NTRK2, and DLG4 could be pivotal genes responsible for triggering apoptosis in hypoxic environments during CI-reperfusion injury.

AIM: Observation of neuroprotective effects of Toddalia asiatica (TA) on cerebral ischemia-reperfusion injury (CIRI) in rats by investigating the effects and mechanisms of drugs on the polarization of microglia M1/M2 subtype and the Toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88)/nuclear factor-κB (NF-κB) pathway. METHODS: The modified thread occlusion method was used to establish a rat model of CIRI, and the rats were randomly divided into the model group, Toddalia asiatica (1.08 g/kg) group, donepezil hydrochloride (0.45 mg/kg) group, and sham group, with 16 rats in each group. Based on the assessment of neurofunctional changes in each group of rats, HE and Nissl staining were used to observe the pathological changes in brain tissue. TUNEL staining was performed to detect neuronal apoptosis, immunohistochemistry was used to detect the expression of M1 microglia marker ionized calcium-binding adapter molecule 1 (Ibal), M2 microglia marker arginase 1 (Argl), and TLR4 localization. Western blot was used to detect the expression of microglia polarization proteins and proteins related to TLR4/MyD88/NF-κB pathway in the hippocampus region. RESULTS: Compared with sham group, the model group rats had higher neurological function scores (P<0.01), and neuronal arrangement in the hippocampus and cortex was loose and disordered, Nissl bodies decreased, and neuronal apoptosis increased. The numbers of M1 microglia marker Iba1-, M2 microglia marker Arg1-, and TLR4-positive cells were significantly increased. In addition, the protein levels of TLR4, MyD88, p-NF-κB p65, NF-κB p65, p-NF-κB inhibitory factor (p-IκB), Ibal, interleukin-6 (IL-6), and Arg1 in the hippocampus were elevated (P< 0.05, while IL-4 and IL-10 expression were decreased (P<0.01). Compared with model group, the Toddalia asiatica group and Donepezil hydrochloride group showed increased protein expression of Arg1, IL-4 and IL-10 (P<0.05), while the other indicators were decreased (P<0.05, P<0.01). CONCLUSION: Toddalia asiatica possesses neuroprotective effects on CIRI rats, which may be attributed to its ability to regulate M1/M2 polarization and inhibit the TLR4/MyD88/NF-κB-mediated inflammatory pathway. [ABSTRACT FROM AUTHOR]

Cerebral stroke has one of the highest incidence, mortality and disability rates worldwide. Although studies have confirmed that reperfusion therapy is an effective strategy for treating stroke patients, the secondary injury of brain tissue caused by reperfusion therapy, which is called cerebral ischaemia reperfusion injury (CIRI), is another medical challenge globally. The microenvironment network constructed by multicellular exosomes plays a regulatory role in the occurrence and development of CIRI. In addition, exosomes have potential value as drug delivery carriers for cerebrovascular diseases due to their stability, the ability to pass through the blood--brain barrier and other characteristics. This study reviews the research progress on the signal network constructed by multiple cells through exosomes and exo-somes as drug carriers in CIRI to provide a reference for research to improve CIRI. [ABSTRACT FROM AUTHOR]

Neuroinflammation and oxidative stress damage are involved in the pathogenesis of cerebral ischemia–reperfusion injury (CIRI). Ferroptosis emerged as a new player in the regulation of lipid peroxidation processes. This study aimed at exploring the potential involvement of ciprofol on ferroptosis-associated CIRI and subsequent neurological deficits in the mouse model of transient cerebral ischemia and reperfusion. Cerebral ischemia was built in male C57BL/6 J wild-type (WT) and Nrf2-knockout (Nrf2 KO) mice in the manner of middle cerebral artery occlusion (MCAO) followed by reperfusion. Ciprofol improved autonomic behavior, alleviated reactive oxygen species output and ferroptosis-induced neuronal death by nucleus transportation of NFE2 like BZIP transcription factor 2 (Nrf2) and the promotion of heme oxygenase 1 (Ho-1), solute carrier family 7 member 11 (SLC7A11/xCT), and glutathione peroxidase 4 (GPX4). Additionally, ciprofol improved neurological scores and reduced infarct volume, brain water content, and necrotic neurons. Cerebral blood flow in MCAO-treated mice was also improved. Furthermore, absence of Nrf2 abrogated the neuroprotective actions of ciprofol on antioxidant capacity and sensitized neurons to oxidative stress damage. In vitro, the primary-cultured cortical neurons from mice were pre-treated with oxygen–glucose deprivation/reperfusion (OGD/R), followed by ciprofol administration. Ciprofol effectively reversed OGD/R-induced ferroptosis and accelerated transcription of GPX4 and xCT. In conclusion, we investigated the ciprofol-induced inhibition effect of ferroptosis-sheltered neurons from lipid preoxidation in the pathogenesis of CIRI via Nrf2-xCT-GPX4 signaling pathway. [ABSTRACT FROM AUTHOR]

This study was to probe the protective effects and mechanisms of salvianolic acid A (SAA) on cerebral ischemia–reperfusion injury (CIRI). The middle cerebral artery occlusion model (MCAO) was established in rats. Rats' behavior, neurological deficits, brain injury, inflammation, and apoptosis in the brain tissue were evaluated. The inflammatory response and apoptosis of PC12 cells induced by oxygen glucose deprivation/reperfusion (OGD/R) were detected. SAA‐mediated changes in miR‐212‐3p, SOX7, and Wnt/β‐catenin pathway were determined, and the targeting relationship between miR‐212‐3p and SOX7 was clarified. SAA alleviated the neurological deficits and brain injury of MCAO rats and inhibited the inflammatory response and apoptosis of OGD/R‐conditioned PC‐12 cells. SAA upregulated miR‐212‐3p, Wnt3a, and β‐catenin, whereas inhibited SOX7 levels. Silencing miR‐212‐3p counteracted the protective effect of SAA in the context of CIRI. SOX7 was a target protein of miR‐212‐3p. Silencing SOX7 based on SAA and miR‐212‐3p knockdown suppressed OGD/R‐induced inflammation and apoptosis and increased Wnt3a and β‐catenin levels in PC12 cells. SAA can improve the brain and nervous system injury caused by cerebral ischemia–reperfusion by upregulating miR‐212‐3p, thereby inhibiting SOX7 and activating the Wnt/βcatenin signaling pathway. [ABSTRACT FROM AUTHOR]

Background and purpose: Cerebral ischemia‒reperfusion injury causes significant harm to human health and is a major contributor to stroke-related deaths worldwide. Current treatments are limited, and new, more effective prevention and treatment strategies that target multiple cell components are urgently needed. Leucine-rich alpha-2 glycoprotein 1 (Lrg1) appears to be associated with the progression of cerebral ischemia‒reperfusion injury, but the exact mechanism of it is unknown. Methods: Wild-type (WT) and Lrg1 knockout (Lrg1−/−) mice were used to investigate the role of Lrg1 after cerebral ischemia‒reperfusion injury. The effects of Lrg1 knockout on brain infarct volume, blood‒brain barrier permeability, and neurological score (based on 2,3,5-triphenyl tetrazolium chloride, evans blue dye, hematoxylin, and eosin staining) were assessed. Single-cell RNA sequencing (scRNA-seq), immunofluorescence, and microvascular albumin leakage tests were utilized to investigate alterations in various cell components in brain tissue after Lrg1 knockout. Results: Lrg1 expression was increased in various cell types of brain tissue after cerebral ischemia‒reperfusion injury. Lrg1 knockout reduced cerebral edema and infarct size and improved neurological function after cerebral ischemia‒reperfusion injury. Single-cell RNA sequencing analysis of WT and Lrg1−/− mouse brain tissues after cerebral ischemia‒reperfusion injury revealed that Lrg1 knockout enhances blood‒brain barrier (BBB) by upregulating claudin 11, integrin β5, protocadherin 9, and annexin A2. Lrg1 knockout also promoted an anti-inflammatory and tissue-repairing phenotype in microglia and macrophages while reducing neuron and oligodendrocyte cell death. Conclusions: Our results has shown that Lrg1 mediates numerous pathological processes involved in cerebral ischemia‒reperfusion injury by altering the functional states of various cell types, thereby rendering it a promising therapeutic target for cerebral ischemia‒reperfusion injury. Highlights: Upregulated Lrg1 expression is observed in various cells in brain after cerebral ischemia‒reperfusion injury. Lrg1 knockout can attenuate brain damage and blood‒brain barrier dysfunction following cerebral ischemia‒reperfusion injury. Lrg1 knockout can alter the functional state of microglial cells towards the anti-inflammatory M2 phenotype. Lrg1 knockout can modulate the metabolic status of multiple cell types. [ABSTRACT FROM AUTHOR]

AIM: To explore the mechanism of Qingnao dripping pills （QNDP） in regulating the nucleotide-binding oligomerization domain-like receptor protein 3 （NLRP3） inflammasome signaling pathway by microRNA-223-3p （miR-223-3p） after acute cerebral ischemia-reperfusion injury in rats. METHODS: The rats with the model of middle cerebral artery occlusion and reperfusion （MCAO/R） were randomly divided into MCAO/R group, MCAO/R+QNDP group, MCAO/R+miR-223-3p inhibitor （antagomiR-223） group, and MCAO/R+QNDP+antagomiR-223 group. Sham group served as a control. There were 18 rats in each group. The QNDP were administered by gavage at 150 mg/kg after MCAO/R, and antagomiR-223 was injected into the lateral ventricle 24 h before MCAO/R operation. The neurological deficit was measured by Garcia score. The volume of cerebral ischemia was evaluated observed by TTC staining. The brain water content was evaluated by dry and wet weight determination. The protein levels of NLRP3, apoptosis-associated speck-like protein containing a caspase recruitment domain （ASC） and cleaved caspase-1 were detected by Western blot. The localization of NLRP3 in the ischemic cortex was observed by immunofluorescence staining. The levels of interleukin-1β （IL-1β） and IL-18 were detected by ELISA. RESULTS: Compared with sham group, the neurological deficit scores were significantly reduced （P<0. 01）, and the cerebral ischemic volume and brain water content were significantly increased （P<0. 01） in MCAO/R model group. Compared with MCAO/R group, the rats in MCAO/R+QNDP group showed that the neurological deficit scores were significantly increased, cerebral ischemic volume and water content were significantly decreased （P<0. 01）, miR-223-3p expression was increased （P<0. 05）, while NLRP3 inflammasome （NLRP3, ASC and cleaved caspase-1） protein expression, NLRP3 positive cell number, and IL-1β and IL-18 levels were decreased（ P<0. 05 or P<0. 01）. The trend of above indicators was consistent with MCAO/R+QDNP+antagomiR-223 group （P<0. 05 or P< 0. 01）, while MCAO/R+antagomiR-223 group showed the opposite trend（ P<0. 05 or P<0. 01）. Compared with MCAO/R+ QNDP group, the rats in MCAO/R+QNDP+antagomiR-223 group showed that neurological function scores and miR-223-3p expression were significantly decreased （P<0. 01）, cerebral infarction volume, brain water content, NLRP3 and cleaved caspase-1 protein expression, and IL-1β and IL-18 levels were increased in the ischemic cortex （P<0. 05 or P< 0. 01）, but ASC protein expression had no statistically difference. Compared with MCAO/R+antagomiR-223 group, ASC protein level was significantly decreased in MCAO/R+QNDP+antagomiR-223 group （P<0. 01）. CONCLUSION: The QNDP may inhibit the NLRP3 inflammasome signaling pathway by up-regulating miR-223-3p expression, thus alleviating the inflammatory response induced by cerebral ischemia-reperfusion injury in rats. [ABSTRACT FROM AUTHOR]

Background: Reperfusion therapy after ischemic cerebral stroke may cause cerebral ischemia-reperfusion injury (CIRI), and cerebral edema is an important factor that may aggravate CIRI. Our study aimed to dynamically monitor the development of early cytotoxic edema after CIRI by magnetic resonance imaging (MRI) and to validate it using multiple histological imaging methods. Methods: Male Sprague Dawley rats were divided into sham and CIRI groups. T2-weighted imaging (T2WI) and diffusion-weighted imaging (DWI)-MRI scans were performed in the sham and CIRI groups after reperfusion. Relative apparent diffusion coefficient (rADC) values were calculated and the midline shift (MLS) was measured. A series of histological detection techniques were performed to observe changes in the cerebral cortex and striatum of CIRI rats. Correlation analysis of rADC values with aquaporin-4 (AQP4) and sodium-potassium-chloride cotransport protein 1 (Na+-K+-2Cl--cotransporter 1; NKCC1) was performed. Results: rADC values began to increase and reached a relatively low value in the cerebral cortex and striatum at 24 h after reperfusion, and the MLS reached relatively high values at 24 h after reperfusion (all p < 0.05). Hematoxylin-eosin (HE) staining showed that the nerve cells in the cortex and striatum of the sham group were regular in morphology and neatly arranged, and in the CIRI-24 h group were irregular, disorganized, and loosely structured. Using terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining, the number of TUNEL+ cells in the ischemic cortex and striatum in CIRI-24 h group was shown to increase significantly compared with the sham group (p < 0.05). Transmission electron microscopy showed that the perivascular astrocytic foot processes were swollen in the cortex and striatum of the CIRI-24 h group. Pearson correlation analysis demonstrated that rADC values were negatively correlated with the number of anti-glial fibrillary acidic protein (GFAP)+AQP4+ and GFAP+NKCC1+ cells of the CIRI rats. Conclusions: MRI combined with histological techniques can dynamically assess cytotoxic edema after CIRI, in a manner that is clear and intuitive for scientific researchers and clinicians, and provides a scientific basis for the application of MRI techniques for monitoring the dynamic progress of CIRI. [ABSTRACT FROM AUTHOR]

Objective To investigate the effects of S100A9 knockdown on cerebral ischemia-reperfusion (I/R) injury and inflammation in rats and to explore the related mechanism. Methods SD rats were randomly divided into the sham operation group, model group, Lsh-NC group, and Lsh-S100A9 group, respectively. The middle cerebral artery occlusion (MCAO) models were prepared by intraluminal suture method in the model group, Lsh-NC group, and LshS100A9 group; the rats in the sham operation group only received the carotid artery exposure and wire embedding; in the Lsh-S100A9 group and Lsh-NC group, rats received S100A9 knockdown lentivirus or empty vector intracerebral injection at 24 h after modeling. RT-qPCR was used to detect S100A9 mRNA expression in the brain tissues of rats; neurological severity score (NSS) score was used to evaluate the degree of neurological impairment; infarct volume of brain tissue was measured by 2, 3, 5-triphenyltetrazolium chloride (TTC) staining; and IL-1β, IL-6 and tumor necrosis factor-α(TNF-α) were detected by ELISA. The protein expression levels of p65, phosphorylated p65(p-p65) and MyD88 in the brain tissues of rats in the Lsh-NC group and Lsh-S100A9 group were detected by Western blotting. Results S100A9 mRNA expression level was as follows: the model group, Lsh-NC group > sham operation group > Lsh-S100A9 group; the neurological impairment score, ischemic cerebral infarction volume and IL-1β, IL-6, TNF-α expression levels in the brain tissues were in the following order: the model group, Lsh-NC group > Lsh-S100A9 group > sham operation group (both P<0. 05). The expression levels of p-p65 and MyD88 protein in the Lsh-S100A9 group were lower than those in the Lsh-NC group (all P<0. 01). Conclusion S100A9 knockdown can alleviate the cerebral I/R injury and inhibit brain inflammation in MCAO rats. The mechanism may be related to the regulation of NF-κB signaling pathway to affect the inflammatory response. [ABSTRACT FROM AUTHOR]

Cerebral ischemia‐reperfusion injury (CIRI) is intimately associated with the redox regulation of biothiol, a crucial antioxidant marker that precludes the onset of ROS. We designed a novel fluorescent probe, DCI‐Ac‐Py, showing various physicochemical properties, such as high selectivity, exceptional signal‐to‐noise ratio, near‐infrared (NIR) optical window, and blood–brain barrier (BBB) penetrability, for detecting biothiols in the brain. The picolinate serves as a specific recognition group that is rapidly activated by biothiol and undergoes nucleophilic substitution with the adjacent acrylic ester to yield the desired NIR probe. Additionally, the probe's lipid solubility is improved through the inclusion of halogen atoms, which aids in penetrating the BBB. Using DCI‐Ac‐Py, we investigated changes of biothiols in vivo in the brains of mice during CIRI. We found that biothiol‐mediated NF‐kB classical (P65‐related) and nonclassical (RelB‐related) pathways contribute to abundant ROS production induced by CIRI and that biothiols are involved in redox regulation. These findings provide new insights into the study of CIRI and shed light on the physiological and pathological mechanisms of biothiols in the brain. [ABSTRACT FROM AUTHOR]

Objective: To explore the role of hypoxia-inducible factor-1α/Bcl-2/adenovirus E1B-19kDa interacting protein 3 （HIF-1α/BNIP3） in IL-4-induced attenuation of cerebral ischemia-reperfusion injury （CIRI） in mice and its relationship with autophagy. Methods: SPF BALB/c mice were divided into sham group, cerebral ischemia reperfusion injury group（CIRI group）, IL-4 group, HIF-1α inhibitor 2ME2 group （2ME2 group） and IL-4+HIF-1α inhibitor 2ME2 group （IL-4+2ME2 group） by random number table method. Middle cerebral artery embolism （MACO） models of CIRI group, IL-4 group, 2ME2 group and IL-4+2ME2 group were constructed by thread embolization method. Occlusion of ischemia was performed for 60 minutes and then reperfusion for 24 hours. In sham-operated group, except that the middle cerebral artery of mice was not blocked, the other operations were the same as above. IL-4 group, 2ME2 group and IL-4+2ME2 group were intraperitoneally injected with the corresponding IL-4C complex solution and the tail vein injection with 2ME2 solution 30 minutes before modeling. After 24 hours of reperfusion, neurobehavioral scores were performed, then the mice was sacrificed and the brain tissues were taken. TTC staining was used to detect cerebral infarction volume; dry and wet weight method was used to determine brain tissue water content; TUNEL staining was used to detect cell apoptosis; transmission electron microscopy was used to count autophagosomes; colorimetric method was used to detect SOD, MDA and ROS levels; Western blot was used to detect expressions of HIF-1α, BNIP3, LC3-Ⅱ and Beclin-1. Results: Compared with sham group, neurobehavioral score, cerebral infarction volume, brain water content, apoptosis rate, number of autophagosomes, and levels of MDA, ROS, HIF-1α, BNIP3, LC3-Ⅱ, Beclin-1 in CIRI group were increased, while level of SOD was decreased （P<0.05）; compared with CIRI group, neurobehavioral score, cerebral infarction volume, brain water content, cell apoptosis rate, MDA and ROS levels in IL-4 group were decreased, the number of autophagosomes, SOD, HIF-1α, BNIP3, LC3-Ⅱ and Beclin-1 levels were increased （P<0.05）; compared with IL-4 group, neurobehavioral score, cerebral infarction volume, brain water content, apoptosis rate, number of autophagosomes, MDA, ROS, HIF-1α, BNIP3, LC3-Ⅱ and Beclin-1 levels in 2ME2 group and IL-4+2ME2 group were decreased, while SOD level was increased （P<0.05）. Conclusion: IL-4 can activate HIF-1α/BNIP3 signaling pathway, activate cell autophagy, reduce cell apoptosis and oxidative stress, thereby alleviating CIRI in mice. [ABSTRACT FROM AUTHOR]

Circular RNAs (circRNAs) are abundantly and stably expressed in the brain of mammals and humans. Some circRNAs are implicated in ischemic stroke. Therefore, we aimed to detect how circRNAs change in the key penumbra area during cerebral ischemia-reperfusion (CI/R) injury. Rats were subjected to transient middle cerebral artery occlusion (tMCAO), during which the permanent blocking period was 2 h and reperfusion time was 24 or 72 h. Then modified neurologic severity score (mNSS), triphenyl tetrazolium chloride (TTC) staining and HE staining were used to exhibiting damage between rats in different groups. The penumbra regions of all rats were dissected and total RNA was further processed for high-throughput sequencing. CircRNA expression profiles were screened and bioinformatics analyses were conducted to investigate these differentially expressed circRNAs. Some of them were verified by reverse transcription-quantitative polymerase chain reaction (RT-qPCR), followed by the establishment of a circRNA-miRNA-mRNA network and the detection of their downstream molecules. A total of 99 and 98 circRNAs were differentially expressed at CI/R 24 h and CI/R 72 h, respectively. Notably, 21 circRNAs significantly changed at both reperfusion points. Three circRNAs, namely circ.7225, circ.5415, and circ.20623 were found to be associated with CI/R injury and might be preferred targets. Common downstream miR-298-5p and Bcl-3 were found to make up the circRNA-miRNA-mRNA network. Novel circRNA targets came to light in the penumbra of rats during CI/R injury and might establish the circRNA-miRNA-mRNA relationship, thus serving as potential biomarkers for ischemic stroke treatment. [ABSTRACT FROM AUTHOR]

Abstract Background Excessive and unresolved neuroinflammation plays an important role in the pathophysiology of many neurological disorders, such as ischemic stroke, yet there are no effective treatments. Tripartite motif-containing 67 (TRIM67) plays a crucial role in the control of inflammatory disease and pathogen infection-induced inflammation; however, the role of TRIM67 in cerebral ischemia‒reperfusion injury remains poorly understood. Results In the present study, we demonstrated that the expression level of TRIM67 was significantly reduced in middle cerebral artery occlusion and reperfusion (MCAO/R) mice and primary cultured microglia subjected to oxygen–glucose deprivation and reperfusion. Furthermore, a significant reduction in infarct size and neurological deficits was observed in mice after TRIM67 upregulation. Interestingly, TRIM67 upregulation alleviated neuroinflammation and cell death after cerebral ischemia‒reperfusion injury in MCAO/R mice. A mechanistic study showed that TRIM67 bound to IκBα, reduced K48-linked ubiquitination and increased K63-linked ubiquitination, thereby inhibiting its degradation and promoting the stability of IκBα, ultimately inhibiting NF-κB activity after cerebral ischemia. Conclusion Taken together, this study demonstrated a previously unidentified mechanism whereby TRIM67 regulates neuroinflammation and neuronal apoptosis and strongly indicates that upregulation of TRIM67 may provide therapeutic benefits for ischemic stroke. Graphical Abstract

Cerebral ischemia-reperfusion injury, one of the main reasons for poor prognosis in patients with neurological diseases, is also an important cause of paralysis. It is more common in cardiac arrest, cerebral hemorrhage, cerebral infarction and stroke. The classical pathophysiological mechanisms mainly include oxidative stress, inflammatory response, calcium overload, nitric oxide and excitatory amino acid toxicity. With the development of medical technology, the treatment of cerebral ischemia-reperfusion injury has been improved. However, there is still an urgent need to explore more accurate and effective treatments. Mesenchymal stem cells (MSCs) are multipotential stem cells in bone marrow, muscle, fat and other tissues and organs. In recent years, MSCs have been explored in the treatment of various diseases because of their strong differentiation ability, secretion ability and good immunocompatibility. This paper describes the pathophysiological process of cerebral ischemia-reperfusion injury, and summarizes the role of MSCs in cerebral ischemia-reperfusion injury from the perspective of pathophysiological mechanism, in order to provide a new idea for the treatment of cerebral ischemia-reperfusion injury.

Wenmei Zhou,1 Tao Tao,1 Wenfeng Yu,2 Wanfu Wu,3 Zhirong Hui,1 Hongliang Xu,1 Yaqi Li,4 Ying Zhang,5 Xiaohui Yang1,6 1Department of Rehabilitation Medicine, Guizhou Provincial People’s Hospital, Guiyang, Guizhou, 550002, People’s Republic of China; 2Department of Human Anatomy, Basic Medical College, Guizhou Medical University, Guiyang, Guizhou, 550004, People’s Republic of China; 3Department of Biology and Biochemistry, University of Houston, Houston, TX, 77204, USA; 4Emergency Department, Guizhou Provincial People’s Hospital, Guiyang, Guizhou, 550002, People’s Republic of China; 5Department of Chinese Traditional Medicine, Zunyi Medical and Pharmaceutical College, Zunyi, Guizhou, 563006, People’s Republic of China; 6Department of Rehabilitation Medicine, The Affiliated Hospital Guizhou Medical University, Guiyang, Guizhou, 550001, People’s Republic of ChinaCorrespondence: Tao Tao, Tel +86 13985162824, Email ttdoc915@sina.comBackground: Hypoxia inducible factor-1α (HIF-1α) regulates glucose metabolism during ischemia. This study investigated the effect of recombinant adenovirus HIF-1ɑ on neurological function and energy metabolism in a rat cerebral ischemia-reperfusion model.Methods: Rats were divided into four groups: sham-operated (Sham) group, cerebral ischemia-reperfusion (CIR) group, recombinant adenovirus empty vector (Ad) group, and recombinant adenovirus-mediated HIF-1α (AdHIF-1α) group. The AdHIF-1α group and the Ad group were injected with AdHIF-1α and Ad in the lateral ventricle. The mNSS was performed at post-ischemia day 0 (P0) and P1, P14 and P28. At P14, the cerebral infarct volume was compared. At P28, HE staining, Nissl stains and TUNEL staining were performed. The expression of HIF-1α, GLUT1 and PFKFB3 were evaluated by Western Blot and immunohistochemistry. High performance liquid chromatography (HPLC) was used to determine the expression of GLUT1 and PFKFB3, and the level of energy metabolites: ATP, ADP and AMP.Results: mNSS scores in the AdHIF-1α group were consistently lower than those in the CIR and Ad groups from P14 (P < 0.05) and Ad groups (P < 0.05). The cerebral infarct volume was reduced in the AdHIF-1α group compared with that in CIR group and Ad group (P < 0.05). At P28, HE showed better pathological changes in AdHIF-1α group. The number of Nissl bodies was increased in the AdHIF-1α group compared with the CIR and Ad groups (P < 0.05). The number of apoptotic cells in the AdHIF-1α group was fewer than that in the CIR and Ad groups (P < 0.05). The expression of HIF-1α, GLUT1 and PFKFB3 was significantly higher in the AdHIF-1α group compared with the CIR and Ad groups (P < 0.05). The ATP, ADP and AMP in the ischemic penumbra were also higher in the AdHIF-1α group (P < 0.05).Conclusion: HIF-lα promoted neurological function recovery and decreased cerebral infarct volume in rats after cerebral ischemia-reperfusion injury by improving energy metabolism.Keywords: cerebral ischemia-reperfusion injury, HIF-lα, energy metabolism, glucose transporter protein-1, GLUT1, 6-phosphofructo-2-kinase/fructose-2, 6-biphosphatase 3, PFKFB3

BACKGROUND: “Tongdu Tiaoshen” electroacupuncture pretreatment is an emerging physiotherapy combining traditional acupuncture with modern electrical stimulation, which has been reported to have significant therapeutic effects on ischemic stroke, but its specific mechanism is still unclear. OBJECTIVE: To investigate the effects of “Tongdu Tiaoshen” electroacupuncture pretreatment on cell apoptosis, the relative expression of microRNA-124 (miR124) in the ischemic penumbra and Notch-1, p-JNK and cysteinyl-aspartate specific protease-3 (caspase-3) proteins in the cortex of rats and their possible mechanisms of action. METHODS: Seventy-five male Sprague-Dawley rats were randomly divided into five groups (n=15 per group): model group, sham-operated group, electroacupuncture pretreatment group, miR-124 antagomir pretreatment group and electroacupuncture+miR-124 agomir pretreatment group. In the electroacupuncture pretreatment group, 20-minute electroacupuncture at Fengchi, Baihui, and Dazhui was performed once a day, for 7 continuous days. In the miR-124 antagomir pretreatment group, miR-124 antagomir was injected into the lateral ventricle of rats on day 7. In the electroacupuncture+miR-124 agomir pretreatment group, miR-124 agomir was injected based on electroacupuncture pretreatment. Model and sham-operated groups were not treated. After the intervention, animal models were created. In the sham-operated group, the rats were only fixed on the operating board and the vessels were blunt-separated. In the other groups, the right cerebral ischemia-reperfusion injury model was established by the middle cerebral artery occlusion method and the Zea Longa suture method. Twenty-four hours after the modeling, the modified neurological severity scoring was performed to determine the damage. TUNEL staining method was then used to measure cell apoptosis in the rat cerebral cortex. Ultrastructure of the rat cerebral cortex was observed under electron microscopy. RESULTS AND CONCLUSION: Compared with the sham-operated group, the modified neurological severity scores were significantly elevated in the other groups (all P < 0.01). Compared with the model group, the miR-124 antagomir pretreatment group and electroacupuncture pretreatment group had significantly reduced modified neurological severity scores (both P < 0.05). Compared with the sham-operated group, the number of apoptotic cells was elevated in the model group (P < 0.01). Compared with the model group, the number of apoptotic cells was reduced in the pretreatment groups (all P < 0.01). The model group had more severe defects in cell structure and contents. The electroacupuncture pretreatment group had mild defects in cell structure and contents. Cells in the miR-124 antagomir pretreatment group were more intact. There was damage to cell structure and contents in the electroacupuncture+miR-124 agomir pretreatment group. The relative expressions of caspase-3, Notch-1 and p-JNK proteins in the cortex of rats were significantly lower in the miR-124 antagomir pretreatment and electroacupuncture pretreatment group compared with the model group (all P < 0.01). The relative expression of miR-124-3p and Notch-1 mRNA in the cortex of the pretreated rats was significantly reduced compared with that of the model group (all P < 0.01). The relative expression of caspase-3 mRNA and p-JNK mRNA in the cortex of rats was significantly reduced in the miR-124 antagomir pretreatment and electroacupuncture pretreatment groups compared with the model group (all P < 0.01). The relative expression of p-JNK mRNA in the cortex of rats in the electroacupuncture+miR-124 agomir pretreatment group was also reduced compared with the model group (P < 0.05). To conclude, “Tongdu Tiaoshen” electroacupuncture pretreatment can inhibit the apoptosis of neuronal cells after cerebral ischemia-reperfusion injury by interfering with the expression level of miR-124 in the brain tissue of rats, thereby reducing neurological deficits after cerebral ischemia-reperfusion injury. The mechanism of this action may be related to the positive regulation mechanism of miR-124 on the Notch pathway. [ABSTRACT FROM AUTHOR]

Background: Cerebral ischemia-reperfusion injury (CIRI) can cause irreversible brain damage and autophagy has been implicated in the pathophysiology. Increasing serum potassium (K+) levels reduces CIRI, but the relationship between its protective mechanism and autophagy is unclear. In this study, we aimed to find the optimal degree of raising serum (K+) and to investigate the relationship between high (K+) and autophagy and the underlying mechanisms in a cardiac arrest/cardiopulmonary resuscitation (CA/CPR) rat model. Methods: Sprague Dawley (SD) rats were divided into four groups: S group, N group, P group, and Q group. The rats S group and N group were administered saline. The rats P group and Q group were administered 640 mg/kg of potassium chloride (KCl) continuously pumped at 4 mL/h (21.3 mg/(kg·min) and divided according to the electrocardiogram (ECG) changes during the administration of KCl. After 24-h of resuscitation, neural damage was assessed by measuring neurological deficit score (NDS), oxidative stress markers, and pathological staining of the cerebral cortex. The level of autophagy and the expression of mTOR-ULK1-Beclin1 pathway-related proteins were evaluated using transmission electron microscopy (TEM), immunostaining, and western blotting. Results: Our results revealed that high (K+) improved NDS and decreased the oxidative stress markers. The autophagosomes, autolysosomes, and lysosomes were decreased following treatment KCl. Furthermore, the levels of micro-tubule-associated protein 1 light chain 3 (LC3) Ⅱ/Ⅰ, Unc-51-like kinase 1 (ULK1), and Beclin1 were decreased, whereas mTOR expression was increased in the cortex. Conclusion: The results demonstrated that moderate hyperkalemia could alleviate autophagy after CIRI via regulating the mTOR-ULK1-Beclin1 pathway. [ABSTRACT FROM AUTHOR]

Background: Ischemic stroke, the most common stroke type, has threatened human life and health. Currently, intravenous thrombolysis and endovascular thrombectomy are the mainstream treatment methods, but they may cause cerebral ischemia-reperfusion injury (CIRI), which aggravates brain injury. Consequently, it is worthwhile to start with a study of CIRI mechanism to identify better prevention and treatment methods. Applying single-cell RNA sequencing (scRNA-seq) technology to further understand the biological functions of various cell types in CIRI will facilitate the intervention of CIRI. Methods: This study aimed to establish a rat middle cerebral artery occlusion (MCAO) model to simulate cerebral ischemia-reperfusion, perform enzymatic hydrolysis, and suspend cerebral cortex tissue edema. Single-cell transcriptome sequencing was used, combined with cluster analysis, t-distributed stochastic neighbor embedding (t-SNE) visualization, and other bioinformatics methods to distinguish cell subgroups while using gene ontology (GO) function enrichment and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment to reveal the biological function of each cell subgroup. Results: We identified 21 brain clusters with cell type-specific gene expression patterns and cell subpopulations, as well as 42 marker genes representing different cell subpopulations. The number of cells in clusters 0-3 increased significantly in MCAO group compared to that in the sham group, and nine-cell subpopulations exhibited remarkable differences in the number of genes. Subsequently, GO and KEGG analyses were performed on the top 40 differentially expressed genes (DEGs) in the six cell subpopulations with significant differences. These results indicate that biological processes and signaling pathways are involved in different cell subpopulations. Conclusions: ScRNA-seq revealed the diversity of cell differentiation and the unique information of cell subpopulations in the cortex of rats with acute ischemic stroke, providing novel insight into the pathological process and drug discovery in stroke. [ABSTRACT FROM AUTHOR]

Inhibition of the inflammatory response triggered by microglial pyroptosis inflammatory activation may be one of the effective ways to alleviate cerebral ischemia–reperfusion injury, the specific mechanism of which remains unclear. In this study, BV-2 microglia with or without oxygen–glucose deprivation/reoxygenation (OGD/R) or long noncoding RNA (lncRNA) Gm44206 knockdown were used as cell models to conduct an in vitro study. Detection of lactate dehydrogenase release and pyroptosis-related protein levels was performed using a corresponding kit and western blotting, respectively. Proliferation of microglia was evaluated by CCK8 assay. Enzyme-linked immunosorbent assay was applied for measuring levels of proinflammatory cytokines. This study verified the involvement of microglial pyroptosis as well as upregulation of NLRP3, Caspase-1, GSDMD, and Apoptosis-associated Speck-like protein containing a C-terminal caspase-recruitment domain (ASC) in cerebral ischemia–reperfusion injury. Moreover, knockdown of lncRNA Gm44206 could alleviate OGD/R-induced microglial pyroptosis and cell proliferation inhibition through the NLRP3/Caspase-1/GSDMD pathway, thus decreasing the release of proinflammatory cytokines, including interleukin (IL)-1β, IL-6, IL-18, and tumor necrosis factor-alpha. In conclusion, this study established a correlation between microglial pyroptosis and cerebral ischemia–reperfusion injury and identified lncRNA Gm44206 as a potential regulator of NLRP3/Caspase-1/GSDMD axis-mediated microglial pyroptosis, which could be considered a promising therapeutic target. [ABSTRACT FROM AUTHOR]

Background: Epigenetic histone methylation plays a crucial role in cerebral ischemic injury, particularly in the context of ischemic stroke. However, the complete understanding of regulators involved in histone methylation, such as Enhancer of Zeste Homolog 2 (EZH2), along with their functional effects and underlying mechanisms, remains incomplete. Methods: Here, we employed a rat model of MCAO (Middle cerebral artery occlusion) and an OGD (Oxygen‐Glucose Deprivation) model of primary cortical neurons to study the role of EZH2 and H3K27me3 in cerebral ischemia‐reperfusion injury. The infarct volume was measured through TTC staining, while cell apoptosis was detected using TUNEL staining. The mRNA expression levels were quantified through quantitative real‐time polymerase chain reaction (qPCR), whereas protein expressions were evaluated via western blotting and immunofluorescence experiments. Results: The expression levels of EZH2 and H3K27me3 were upregulated in OGD; these expression levels were further enhanced by GSK‐J4 but reduced by EPZ‐6438 and AKT inhibitor (LY294002) under OGD conditions. Similar trends were observed for mTOR, AKT, and PI3K while contrasting results were noted for UTX and JMJD3. The phosphorylation levels of mTOR, AKT, and PI3K were activated by OGD, further stimulated by GSK‐J4, but inhibited by EPZ‐6438 and AKT inhibitor. Inhibition of EZH2 or AKT effectively counteracted OGD‐/MCAO‐induced cell apoptosis. Additionally, inhibition of EZH2 or AKT mitigated MCAO‐induced infarct size and neurological deficit in vivo. Conclusions: Collectively, our results demonstrate that EZH2 inhibition exerts a protective effect against ischemic brain injury by modulating the H3K27me3/PI3K/AKT/mTOR signaling pathway. The results provide novel insights into potential therapeutic mechanisms for stroke treatment. [ABSTRACT FROM AUTHOR]

Alterations in the M1/M2 polarization phenotype significantly affect disease progression. Antioxidant and anti-inflammatory protective effects of resveratrol (Res) have been demonstrated. This paper tested the hypothesis that Res could protect against cerebral ischemia–reperfusion injury (CI/RI) by modulating microglial polarization via the miR-450b-5p/KEAP1/Nrf2 pathway. Rats were first treated with Res and adenovirus that interfered with miR-450b-5p or KEAP1, and then established a middle cerebral artery occlusion-reperfusion model using modified nylon sutures. Rats were then evaluated for neurological and behavioral functions, and markers of M2 microglia were detected by immunofluorescence staining. Additionally, the signature patterns of miR-450b-5p, KEAP1, and Nrf2 were determined. The collected data demonstrated that Res exerted neuroprotective effects in CI/RI by promoting microglial M2 polarization. Additionally, Res could regulate the Nrf2 pathway by targeting KEAP1 by up-regulating miR-450b-5p. Up-regulating miR-450b-5p or down-regulating KEAP1 could further promote the protective effect of Res, while down-regulating miR-450b-5p or up-regulating KEAP1 worked oppositely. Our study demonstrates that Res exerts neuroprotective effects on microglial M2 polarization through the miR-450b-5p/KEAP1/Nrf2 pathway during CI/RI. [ABSTRACT FROM AUTHOR]

Cerebral ischemia-reperfusion (CIR) injury is initiated by the generation of reactive oxygen species (ROS), which leads to the oxidation of cellular proteins, DNA, and lipids as an initial event. The reperfusion process impairs critical cascades that support cell survival, including mitochondrial biogenesis and antioxidant enzyme activity. Failure to activate prosurvival signals may result in increased neuronal cell death and exacerbation of CIR damage. Melatonin, a hormone produced naturally in the body, has high concentrations in both the cerebrospinal fluid and the brain. However, melatonin production declines significantly with age, which may contribute to the development of age-related neurological disorders due to reduced levels. By activating various signaling pathways, melatonin can affect multiple aspects of human health due to its diverse range of activities. Therefore, understanding the underlying intracellular and molecular mechanisms is crucial before investigating the neuroprotective effects of melatonin in cerebral ischemia-reperfusion injury. [ABSTRACT FROM AUTHOR]

Objective To investigate the effects of baicalin on neuroinflammation and cognitive dysfunction in mice with cerebral ischemia-reperfusion injury (CIRI) by inhibiting PGE2. Methods Adult male C57BL/ 6J mice were randomly divided into two groups, the model of bilateral carotid artery occlusion (tBCCAO ) was established. Baicalin was given by gavage or PGE2 was injected into lateral ventricle.The cognitive function of mice was detected by Morris water maze. The lipid peroxidation and neuroinflammation in hippocampus were measured by ELISA at 72 h after operation. The pathological changes of brain tissue were observed by HE staining. Western blot was used to detect the expression of GPX4, DMT1 and PTGS2 in hippocampus. Results CIRI induced cognitive impairment in mice, accompanied by hippocampal neuronal injury, neuroinflammation, lipid peroxidation and abnormal expression of ferroptosis associated proteins (P < 0.05). Baicalin pretreatment could improve the cognitive impairment induced by CIRI in mice, and was accompanied by a reduction in hippocampal neuronal injury, neuroinflammation, lipid peroxidation and abnormal expression of ferroptosis associated proteins (P < 0.05).PGE2 was upregulated in the hippocampus of mice after intracerebroventricular injection of PGE2, accompanied by neuroinflammation, lipid peroxidation and abnormal expression of ferroptosis associated proteins (P < 0.05). Baicalin could decrease the neuroinflammation, lipid peroxidation and abnormal expression of ferroptosis associated proteins in hippocampus by inhibiting PGE2 up-regulation (P < 0.05). The up-regulation of PGE2 in hippocampus aggravated the cognitive impairment induced by CIRI (P < 0.05). Baicalin improved cognitive impairment by inhibiting the up-regulation of PGE2 (P < 0.05 ). Conclusion Baicalin ameliorates the cognitive impairment induced by CIRI, and the mechanism is related to the inhibition of PGE2-related neuroinflammation. [ABSTRACT FROM AUTHOR]

• PGK1 promotes the progress of cerebra ischemia–reperfusion injury (CIRI). • Nrf2 is a target of PGK1. • Knockdown of Nrf2 abolishes the effect of CBR-470-1 (a PGK1 inhibitor) on CIRI. • PGK1 aggravates CIRI by inhibiting Nrf2/ARE pathway. Understanding the role and mechanism of astrocytes in inflammation and oxidative response is crucial for developing therapeutic strategies to reduce inflammation and oxidative injury in cerebral ischemia–reperfusion injury (CIRI). In this study, we investigated the regulatory effects of phosphoglycerate kinase 1 (PGK1) on inflammation and oxidative response after CIRI in male adult Sprague-Dawley (SD) rats and using primary astrocytes obtained from neonatal SD rats, and explored its related mechanisms. We established a rat model of middle cerebral artery occlusion-reperfusion (MCAO/R) by suture occlusion, and an oxygen-glucose deprivation/reoxygenation model of astrocytes using oxygen-free, glucose-free, and serum-free cultures. AAV8-PGK1-GFP was injected into the left ventricle 24 h before modeling. Real-time quantitative polymerase chain reaction, enzyme-linked immunosorbent assay, co-immunoprecipitation (CoIP) assay, fluorescence in situ hybridization (FISH), and western blotting were used to elucidate the in-depth mechanisms of PGK1 in CIRI. PGK1 overexpression significantly exacerbated neurological deficits, increased cerebral infarct volume, and aggravated nerve cell injury in rats after MCAO/R. Using FISH and CoIP assays, we verified the localization of PGK1 and Nrf2 in primary astrocytes. Further rescue experiments showed that Nrf2 knockdown eliminated the protective effect of CBR-470-1 (a PGK1 inhibitor) on CIRI. Lastly, we confirmed that PGK1 aggravates CIRI by inhibiting the Nrf2/ARE pathway. In conclusion, our findings suggest that inhibiting PGK1 attenuates CIRI by reducing the release of inflammatory and oxidative factors from astrocytes by activating the Nrf2/ARE signaling pathway. [ABSTRACT FROM AUTHOR]

Ferroptosis is a newly described form of regulated necrotic cell death, which is engaged in the pathological cell death related to stroke, contributing to cerebral ischemia-reperfusion (I/R) injury. Therefore, we performed this study to clarify the role of GATA6 in neuronal autophagy and ferroptosis in cerebral I/R injury. The cerebral I/R injury-related differentially expressed genes (DEGs) as well as the downstream factors of GATA6 were predicted bioinformatically. Moreover, the relations between GATA6 and miR-193b and that between miR-193b and ATG7 were evaluated by chromatin immunoprecipitation and dual-luciferase reporter assays. Besides, neurons were treated with oxygen-glucose deprivation (OGD), followed by overexpression of GATA6, miR-193b, and ATG7 alone or in combination to assess neuronal autophagy and ferroptosis. At last, in vivo experiments were performed to explore the impacts of GATA6/miR-193b/ATG7 on neuronal autophagy and ferroptosis in a rat model of middle cerebral artery occlusion (MCAO)-stimulated cerebral I/R injury. It was found that GATA6 and miR-193b were poorly expressed in cerebral I/R injury. GATA6 transcriptionally activated miR-193b to downregulate ATG7. Additionally, GATA6-mediated miR-193b activation suppressed neuronal autophagy and ferroptosis in OGD-treated neurons by inhibiting ATG7. Furthermore, GATA6/miR-193b relieved cerebral I/R injury by restraining neuronal autophagy and ferroptosis via downregulation of ATG7 in vivo. In summary, GATA6 might prevent neuronal autophagy and ferroptosis to alleviate cerebral I/R injury via the miR-193b/ATG7 axis. [ABSTRACT FROM AUTHOR]

AIM: To explore the mechanism of Buyang-Huanwu decoction in alleviating cerebral ischemia-reperfusion injury. METHODS: Rat astrocyte CTX TNA2 cells at the logarithmic growth stage were randomly divided into 5 groups: blank control group (CON group), model group ［(oxygen-glucose deprivation/reoxygenation (OGD/R) group］, Buyang-Huanwu decoction-medicated serum (BYHWDS) group, miR-148a-3p inhibitor group (inhibitor group), and BYHWDS+inhibitor group. All experiments were conducted three times with three replicates. Apart from the cells in CON group, the cells in the other groups were subjected to OGD for 6 h and reoxygenation for 24 h, and were then treated with the drugs at the same time of reoxygenation (transfection was performed before modeling). Cell morphology was observed by inverted phase-contrast microscopy and viability was measured by MTS method. The expression of microRNA-148a-3p (miR-148a-3p), glial fibrillary acidic protein (GFAP) mRNA and caspase-3 mRNA was measured by qRT-PCR, and the protein levels of GFAP, Bcl-2, Bax, caspase-3, cleaved caspase-3, caspase-9 and cleaved caspase-9 were assessed by Western blot. Apoptosis was measured using an annexin V-FITC/PI kit. The expression of GFAP, caspase-3, Bcl-2 and Bax was also evaluated by immunofluorescence double-staining. RESULTS: Compared with CON group, the cells in OGD/R and inhibitor groups were observed to be shrunken and deformed, with increased cell debris and significantly re⁃ duced viability (P<0. 01). The expression of miR-148a-3p was significantly decreased (P<0. 01), while the mRNA levels of GFAP and caspase-3 were significantly increased (P<0. 01). The protein expression of GFAP, the cleaved caspase-3/ caspase-3, Bax/Bcl-2 and cleaved caspase-9/caspase-9 ratios, and the apoptosis level were significantly increased (P< 0. 05). Compared with OGD/R and inhibitor groups, the cell morphology of BYHWDS and BYHWDS+inhibitor groups re⁃ covered after injury, with reduced atrophy, increased connections, and significantly increased cell viability (P<0. 01). The expression of miR-148a-3p was significantly increased (P<0. 01), and the mRNA levels of GFAP and caspase-3 were significantly decreased (P<0. 01). The protein expression of GFAP and apoptosis-related proteins, and the apoptosis level were significantly decreased (P<0. 05). CONCLUSION: Buyang-Huanwu decoction activates miR-148a-3p to reduce the expression of its target gene GFAP, thus reducing apoptosis of rat astrocyte CTX TNA2 cells with OGD/R, alleviating cerebral ischemia-reperfusion injury, and playing a neuroprotective role. [ABSTRACT FROM AUTHOR]

Cerebral ischemia-reperfusion (CIR) injury occurs as a secondary injury during the treatment of ischemic stroke (IS). There is a high death rate and morbidity due to IS throughout the world. Even though Naoxintong Capsule (NXT) is effective in the treatment of CIR, its mechanisms of action are unclear. The study aims to explore the clear mechanism associated with NXT therapy for CIR. We established the model of middle cerebral artery occlusion to evaluate the neurological function and assess the infarct size. Brain tissue metabolomics was used to identify different metabolites, and metabolic profiling systems enriched metabolic pathways. Then, the potential targets of NXT in the treatment of CIR were explored by proteomic, transcriptomic, and metabolomic methods. NXT improves CIR symptoms. We found potential 11 proteins and corresponding metabolites involved in NXT treatment of CIR. Most of these metabolites are regulated to restore after treatment. According to network pharmacology, we found 6 hub genes, including Glb1, Gmps, Pfas, Atic, Gaa, and Acox1, and their associated core metabolites and pathways. This study reveals the complex mechanism of NXT in treating CIR, and provides a new strategy for future researchers to screen related targets and pathways. [ABSTRACT FROM AUTHOR]

Cerebral ischemia-reperfusion injury (CIRI) is a complex pathological condition with high mortality. In particular, reperfusion can stimulate overproduction of reactive oxygen species (ROS) and activation of inflammation, causing severe secondary injuries to the brain. Despite tremendous efforts, it remains urgent to rationally design antioxidative agents with straightforward and efficient ROS scavenging capability. Herein, a potent antioxidative agent was explored based on iridium oxide nano-agglomerates (Tf-IrO 2 NAs) via the facile transferrin (Tf)-templated biomineralization approach, and innovatively applied to treat CIRI. Containing some small-size IrO 2 aggregates, these NAs possess intrinsic hydroxyl radicals (•OH)-scavenging ability and multifarious enzyme activities, such as catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GP x). Moreover, they also showed improved blood-brain barrier (BBB) penetration and enhanced accumulation in the ischemic brain via Tf receptor-mediated transcytosis. Therefore, Tf-IrO 2 NAs achieved robust in vitro anti-inflammatory and cytoprotection effects against oxidative stress. Importantly, mice were effectively protected against CIRI by enhanced ROS scavenging activity in vivo , and the therapeutic mechanism was systematically verified. These findings broaden the idea of expanding Ir-based NAs as potent antioxidative agents to treat CIRI and other ROS-mediated diseases. (1) The ROS-scavenging activities of IrO 2 are demonstrated comprehensively, which enriched the family of nano-antioxidants. (2) The engineering Tf-IrO 2 nano-agglomerates present unique multifarious enzyme activities and simultaneous transferrin targeting and BBB crossing ability for cerebral ischemia-reperfusion injury therapy. (3) This work may open an avenue to enable the use of IrO 2 to alleviate ROS-mediated inflammatory and brain injury diseases. [Display omitted] [ABSTRACT FROM AUTHOR]

Cerebral ischemic stroke (CIS) has become the second leading cause of death worldwide, which is largely related to cerebral ischemia reperfusion injury (CIRI). Surgical intervention is a reliable treatment for CIS, which predictably causes cerebral reperfusion. Therefore, the choice of anesthetic drugs has important clinical significance. Isoflurane (ISO), one of the most used anesthetics, attenuates cognitive impairment and has brain protective effects. However, the role of isoflurane in regulating autophagy and its regulatory mechanism on inflammation in CIRI are still unclear. The middle cerebral artery occlusion (MCAO) method was used to establish a rat model of CIRI. After 24 h of reperfusion, all rats were evaluated by mNSS scoring and dark avoidance experiment. Western blotting and immunofluorescence were used to examine the expression of key proteins. Compared with the sham group, the MCAO group showed increased neurobehavioral scores and decreased cognitive memory function (P < 0.05). As for the ISO-treated MCAO rats, the neurobehavioral score was significantly decreased, the expression of AMPK, ULK1, Beclin1, and LC3B was significantly increased, and the cognitive and memory functions were also significantly improved (P < 0.05). After inhibition of autophagy pathway or key protein AMPK in autophagy, neurobehavioral scores and protein expression of NLRP3, IL-1β, and IL-18 were significantly increased (P < 0.05). Isoflurane post-treatment may enhance autophagy by activating the AMPK/ULK1 signaling pathway and further inhibit the release of inflammatory factors from NLRP3 inflammasomes, thereby ameliorating neurological function and cognitive impairment and exerting a protective effect on the brain in CIRI rats. [ABSTRACT FROM AUTHOR]

Cerebral ischemia--reperfusion injury (CIRI) is a series of cascade reactions that occur after blood flow recanalization in the ischemic zone in patients with cerebral infarction, causing an imbalance in intracellular homeostasis through multiple pathologies such as increased oxygen free radicals, inflammatory response, calcium overload, and impaired energy metabolism, leading to mitochondrial dysfunction and ultimately apoptosis. Rescue of reversibly damaged neurons in the ischemic hemispheric zone is the key to saving brain infarction and reducing neurological deficits. Complex and active neurological functions are highly dependent on an adequate energy supply from mitochondria. Mitochondrial biogenesis (MB), a process that generates new functional mitochondria and restores normal mitochondrial function by replacing damaged mitochondria, is a major mechanism for maintaining intra-mitochondrial homeostasis and is involved in mitochondrial quality control to ameliorate mitochondrial dysfunction and thus protects against CIRI. The main regulator of MB is peroxisome proliferator-activated receptor gamma coactivator-1α (PGC-1α), which improves mitochondrial function to protect against CIRI by activating its downstream nuclear respiratory factor 1 (NRF1) and mitochondrial transcription factor A (TFAM) to promote mitochondrial genome replication and transcription. This paper provides a theoretical reference for the treatment of neurological impairment caused by CIRI by discussing the mechanisms of mitochondrial biogenesis during cerebral ischemia--reperfusion injury. [ABSTRACT FROM AUTHOR]

Objective To observe the therapeutic effect of curcumin on cerebral ischemia-reperfusion injury rats and to explore its mechanism. Methods Sixty Sprague-Dewley （SD) rats were randomly divided into the curcumin group, model group, and sham group, with 20 rats in each group. Rats in the curcumin group and model group were treat‐ ed with thread occlusion method to make middle cerebral artery occlusion （MCAO) models in rats. After successful model‐ ing, curcumin （200 mg/kg) and the same amount of saline solution were injected intraperitoneally, respectively, for 6 con‐ secutive days. In the sham group, only the common carotid artery and external carotid artery were stripped, and no arteries were ligation and no thread plugs were inserted. Bederson score was used to evaluate the severity of neurological impair‐ ment in rats, and TTC staining was used to observe and calculate the percentage of infarct volume in the brain tissues. Per‐ oxisome proliferator-activated receptor γ（PPAR-γ), apoptosis-related blotch-like protein （ASC), and Caspase-1 protein in the brain tissues were detected by Western blotting. Plasma tumor necrosis factor-α（TNF-α) and interleukin-1β（IL1β) were detected by ELISA. Results On the 1st and 7th days after modeling, the neurological deficit scores in the mod‐ el group and curcumin group were higher than those in the sham group （P<0. 05). On the 7th day after modeling, the neurological deficit score of the curcumin group was lower than that of the model group （P<0. 05). The percentage of cerebral infarction volume in the model group was higher than that in the sham group, and the percentage of cerebral infarction volume in the curcumin group was lower than that in the model group （all P<0. 05). The expression of PPAR-γ protein in the brain tissues of the model group was lower than that of the sham group, and the expression of ASC, Caspase-1 protein and plasma levels of TNF-α and IL-1β were higher than those of the sham group （all P<0. 05). The expression of PPAR-γ protein in the brain tissues of the curcumin group was higher than that of the model group, and the expression of ASC, Cas‐ pase-1 protein and plasma levels of TNF-α and IL-1β were lower than those of the model group （all P<0. 05). Conclusion Curcumin can protect the nerve function and reduce the cerebral infarct volume in rats with cerebral ischemireper‐ fusion injury, and its mechanism may be related to regulating the expression levels of PPAR-γ, ASC, Caspase-1, and down-regulating the levels of TNF-α and IL-1β. [ABSTRACT FROM AUTHOR]

Ischemic stroke is a refractory disease that endangers human health and safety owing to cerebral ischemia. Brain ischemia induces a series of inflammatory reactions. Neutrophils migrate from the circulatory system to the site of cerebral ischemia and accumulate in large numbers at the site of inflammation across the blood–brain barrier. Therefore, hitchhiking on neutrophils to deliver drugs to ischemic brain sites could be an optimal strategy. Since the surface of neutrophils has a formyl peptide receptor (FPR), this work modifies a nanoplatform surface by the peptide cinnamyl‐F‐(D)L‐F‐(D)L‐F (CFLFLF), which can specifically bind to the FPR receptor. After intravenous injection, the fabricated nanoparticles effectively adhered to the surface of neutrophils in peripheral blood mediated by FPR, thereby hitchhiking with neutrophils to achieve higher accumulation at the inflammatory site of cerebral ischemia. In addition, the nanoparticle shell is composed of a polymer with reactive oxygen species (ROS)‐responsive bond breaking and is encased in ligustrazine, a natural product with neuroprotective properties. In conclusion, the strategy of hitching the delivered drugs to neutrophils in this study could improve drug enrichment in the brain, thereby providing a general delivery platform for ischemic stroke or other inflammation‐related diseases. [ABSTRACT FROM AUTHOR]