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This study aimed to observe the effects of acupuncture on iron metabolism in the hippocampus of rats with cerebral ischemia-reperfusion injury (CIRI) and to explore the mechanism of acupuncture on neural repair in CIRI rats. After general feeding for 3 days, 16 of 48 healthy Sprague–Dawley rats were selected according to a random number table to form a sham-occlusion group, and the models were prepared in the remaining rats. After successful modeling, the rats were randomized into model and acupuncture groups, with 16 rats in each group. The CIRI models were prepared using the modified Zea Longa suture-occlusion method. In the acupuncture group, the rats were fixed and acupuncture was delivered at "Shuigou" (GV26), "Dazhui" (GV14), and "Baihui" (GV20), and the needles were manipulated once every 15 min. In the sham-occlusion and model groups, rats were fixed without acupuncture. The intervention was performed for 30 min in each group, once every 12 h, seven times. The degree of neurological impairment was assessed by using the modified Garcia score. Cerebral blood flow (CBF) was monitored using a laser speckle blood flow imaging system. The cerebral infarction area ratio was measured using triphenyltetrazolium chloride staining. Ferric ion aggregation in brain tissue was noted using Prussian blue staining. The ferric ion and glutathione (GSH) contents of the hippocampal tissue on the ischemic side were detected using ferric ion and GSH kits. The protein expression of transferrin receptor 1 (TFR1), iron regulatory protein 2 (IRP2), and ferroportin (FPN) was detected via western blotting. (1) Before intervention, compared with the sham-occlusion group, the Garcia neurological score was significantly reduced in the model and acupuncture groups (both P < 0.01). After the intervention, compared with the sham-occlusion group, the Garcia neurological score in the model group decreased significantly (P < 0.01), and the score in the acupuncture group was elevated compared with the model group (P < 0.05). Compared to before the intervention, the Garcia neurological score increased after the intervention in the acupuncture group (P < 0.05). (2) Compared with the sham-occlusion group, in the model group, CBF was reduced, the infarct area was significantly enlarged, ferric ion aggregation increased in the hippocampus, the content of ferric ion increased, and that of GSH decreased significantly (both P < 0.01), while the protein expression of TFR1 and IRP2 was upregulated and that of FPN was downregulated (both P < 0.05). When compared with the model group, in the acupuncture group, CBF was increased, the infarct area was reduced (P < 0.01), the ferric ion aggregation was reduced in the hippocampus, the content of ferric ion decreased, and that of GSH increased significantly (P < 0.05, P < 0.01); the protein expression of TFR1 and IRP2 was downregulated and that of FPN was upregulated (both P < 0.05). Acupuncture can alleviate CIRI, which may be related to the regulation of iron homeostasis-associated transferrin in the hippocampus, attenuation of ferric ion aggregation in hippocampal tissue, reduction of lipid peroxidation, and inhibition of ferroptosis. [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]

To observe the changes in the neurological score, infarct size, and autophagy- and apoptosis-related indicators following the acupuncture treatment of rats with cerebral ischemia-reperfusion injury (CIRI) in order to investigate the effects of acupuncture on autophagy and apoptosis in the hippocampal tissue of CIRI rats In this study, 170 Sprague-Dawley rats were randomly assigned to the normal group (n=34), sham group (n=34), and model establishment group (n=102). A CIRI rat model was established in the model establishment group. The rats of the model establishment group were then randomly and equally allocated to the middle cerebral artery occlusion/reperfusion group (MCAO/R group), acupuncture group (AC group), and edaravone group (ED group) (n=34 each). Each group was further subdivided into the 24-h and 72-h subgroups (n=17 each). The rats of the MCAO/R group were immobilized for 30 min and administered with an intraperitoneal injection of 0.9% sodium chloride solution. The rats of the AC group were immobilized, and they received acupuncture at the"Dà zhuī (大椎 GV 14) ","Shuĭ gōu (水沟 GV 26) ", and "Băi huì (百会 GV 20) " and an intraperitoneal injection of 0.9% sodium chloride solution, with the acupuncture needles left in place for 30 min. The rats of the ED group were immobilized for 30 min and administered with an intraperitoneal injection of edaravone (5 mg/kg). All interventions were performed once every 12 h. Investigations of the neurological score, infarct size using triphenyl tetrazolium chloride staining, hippocampal Caspase-3 protein expression by immunohistochemical assay, hippocampal Beclin-1 expression by Western blotting, autophagosomes in the hippocampus by transmission electron microscopy, and apoptotic rate in the brain tissue of the ischemic side using the terminal deoxynucleotidyl transferase dUTP nick end labeling assay were performed both before and after the interventions. Before intervention, the neurological score of the model establishment group was significantly lower than that of the normal and sham groups (P <0.01). After 24 h and 72 h, the neurological scores of the ED and AC groups were significantly increased (P <0.01), whereas the infarct size ratio were smaller (P <0.05 or P <0.01) and the apoptotic rates and Caspase-3 expression were lower (P <0.01) when compared with the MCAO/R group. The hippocampal Beclin-1 expression in the ED and AC groups was downregulated after 24 h of intervention (P <0.05) but upregulated after 72 h of intervention (P <0.05 and P <0.01, respectively). In the ED and AC groups, 72 h of intervention led to significant increases in the neurological score (P <0.01) and significant reductions in the apoptotic rate and Caspase-3 expression (P <0.05 or P <0.01), but no significant changes in the Beclin-1 expression (P >0.05) when compared with 24 h of intervention. Moreover, the AC group showed a significant decrease in the infarct size ratio (P <0.01). Electron microscopy revealed that after 24 h of intervention, only individual lysosome structures could be observed in the tissues of the AC and ED groups. After 72 h of intervention, co-existence of autophagosomes and lysosomes was observed in the ED group, whereas co-existence of lysosomes and autolysosomes was observed in the AC group. Acupuncture significantly improved the neurological score and reduced the infarct size in the CIRI rats. The mechanisms of these effects could be attributed to the regulation of autophagy and inhibition of apoptosis in the hippocampal tissue as well as the duration of the intervention to a certain extent. [ABSTRACT FROM AUTHOR]

Gastrodia elata (GE) has been widely used in clinical practice for many years with the functions of relieving stroke, suppressing liver Yang, dispelling wind and clearing collaterals. Our group's previous experimental studies have proved that GE has therapeutic effect on cerebral ischemia reperfusion injury (CIRI) (Ding et al., 2022). However, the active components of GE in treating CIRI remain unclear and require further research. The purpose of this paper was to explore the potential effective components of GE improving CIRI based on the "Spectrum-effect" correlation. Zebrafish model was used for verification in vivo experimental. First, the absorption components and metabolites of GE in rat serum were identified using ultra-high performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UHPLC-Q-TOF/MS). Second, pharmacodynamic indexes were determined by ELISA kit method, and the effect-time curve of each pharmacodynamic indexes was established. The potential compounds were screened using the statistical method of grey correlation between pharmacodynamic indicator and component response. Finally, the zebrafish CIRI model was successfully established, and the in vivo effect of the active components of GE was verified intuitively. 45 chemical components were detected in GE. A total of 87 active components in serum of GE were identified including 25 prototype components and 62 metabolites. GE can improve CIRI by regulating the levels of interleukin-6 (IL-6), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α), MDA levels and SOD levels. It was found that p‑hydroxy benzaldehyde (PHB), p-hydroxybenzyl alcohol (PHBA) and gastrodin (GA) of GE were the possibly main active components by grey correlation statistics. The in vivo experiments of zebrafish model showed that PHB, PHBA, and GA have the ability to ameliorate cerebral thrombosis by regulation of oxidative stress and apoptosis. The potential active components of GE on CIRI were initially excavated using UHPLC-Q-TOF-MS/MS, pharmacodynamics, and in vivo experiments of zebrafish model. It makes up for the disadvantages of separate research on chemical components and pharmacodynamics, and reflects the material basis of pharmacodynamics more objectively. It has provided theoretical basis for further quality evaluation and scientific foundation for rational drug using of GE in clinical. [Display omitted] [ABSTRACT FROM AUTHOR]

• CIRI induces metabolic imbalance in brain microregions, tissues, and systemic circulation. • AC demonstrates significant therapeutic effects on CIRI. • AFADESI-MSI efficiently visualized brain microregion metabolism. • A novel research strategy has been devised to unveil metabolic regulatory mechanisms. The combination of Astragalus membranaceus and Carthamus tinctorius (AC) exhibits significant therapeutic effects in cerebral ischemia/reperfusion injury (CIRI). Understanding the metabolic characteristics of brain microregions and disturbances in tissues and systemic circulation is crucial for elucidating the mechanisms of CIRI and the therapeutic benefits of AC. However, in situ metabolic regulation of the complex brain structure has not been adequately studied, and the therapeutic mechanism of AC requires immediate clarification. The present study aimed to unveil the specific metabolic reprogramming of CIRI at systemic and microregional levels, identify key metabolic pathways and metabolites, and elucidate the therapeutic mechanisms of AC. Air flow-assisted desorption electrospray ionization mass spectrometry imaging (AFADESI-MSI), a newly developed technique, was used to investigate metabolites in brain microregions. Hematoxylin-eosin, Nissl, and immunofluorescence staining were performed to visualize the microscopic changes associated with spatial metabolism. A comprehensive metabolomics study was conducted on serum, brain tissue, and microregions, along with neurological assessments, cerebral infarction measurements, and Evans blue experiments, to assess the systemic and local metabolic effects of AC treatment for CIRI. AC significantly reduced neurological damage, minimized infarct size, and repaired blood–brain barrier damage in CIRI rats. AFADESI-MSI demonstrated that the metabolic imbalance caused by CIRI primarily occurs in the cerebral cortex, hippocampus, caudate putamen, thalamus, cerebellar cortex, and fiber tract regions. Significant changes in 16 metabolites were observed in these regions, corresponding to neuron damage, glial cell activation, and neural repair. 20 metabolites from serum and 4 from brain tissue varied significantly with the sham group. Comprehensive metabolomics analysis indicated a close relationship among serum, tissue, and microregional metabolism. CIRI-induced systemic and localized metabolic disorders involve 14 metabolic pathways. AC conferred therapeutic benefits in CIRI by reversing various metabolic imbalances. AFADESI-MSI efficiently visualized brain microregion metabolism. Comprehensive metabolomics analysis revealed detailed insights into the specific metabolic reprogramming in CIRI and the therapeutic impacts of AC. AC demonstrated significant clinical potential as an adjunct therapy to existing CIRI treatments. [Display omitted] [ABSTRACT FROM AUTHOR]

To explore the mechanism of Maresin1 in reducing cerebral ischemia-reperfusion injury. Male C57BL/6 mice were randomly divided (n = 5 in each group), and focal middle cerebral artery occlusion (MCAO) model was used to simulate cerebral ischemia/reperfusion injury. TTC and the Longa score were used to detect the degree of neurological deficits. Western blot was used to detect the expression levels of GSDME, GSDME-N, caspase-3 and cleaved caspase-3 in cerebral ischemic penumbra tissue, and immunofluorescence was used to detect the expression levels of GSDME-N. The mRNA expression levels of GSDME and pro-inflammatory cytokines (IL-1β, IL-6 and TNF-α) were detected by RT-PCR. Compared with sham group, GSDME mRNA levels in MCAO group were significantly increased at 12 h and 24 h after reperfusion, and GSDME and GSDME-N significantly increased at 6–48 h after reperfusion. Compared with sham group, the percentage of infarct size, the Longa score, the mRNA expression levels of IL-1β, IL-6 and TNF-α, and GSDME, GSDME-N, caspase-3 and cleaved caspase-3 in MCAO group was significantly increased. Then, the percentage of infarct size and the Longa score significantly decreased after MaR1 administration, the mRNA expression levels of IL-1β and IL-6 downregulated, and GSDME, GSDME-N, caspase-3 and cleaved caspase-3 were also reduced. After administration of Z-DEVD-FMK(ZDF), the expression of caspase-3, cleaved caspase-3 and GSDME-N was decreased, which in MCAO+MaR1+ZDF group was not statistically significant compared with MCAO+ ZDF group. Maresin1 alleviates cerebral ischemia/reperfusion injury by inhibiting pyroptosis mediated by caspase-3/GSDME pathway and alleviating neuroinflammation. [ABSTRACT FROM AUTHOR]

Cerebral ischemia-reperfusion injury (CIRI) is a destructive adverse reaction of ischemic stroke, leading to high disability and mortality rates. Salvia miltiorrhiza Bge. (Danshen, DS) processed with porcine cardiac blood (PCB-DS), a characteristic processed product, has promising anti-ischemic effects. However, the underlying mechanism of PCB-DS against CIRI remains unclear. Ferroptosis is demonstrated to be involved in CIRI. The aim of this study was to explore the molecular mechanism underlying PCB-DS inhibited GLRX5-mediated ferroptosis alleviating CIRI, which was different from DS. Quality evaluation of PCB-DS and DS was conducted by UPLC. Pharmacological activities of PCB-DS and DS against CIRI were compared using neurobehavioral scores, infarct volume, proinflammatory factors, and pathological examinations. Proteomics was employed to explore the potential specific mechanism of PCB-DS against CIRI, which was different from DS. Based on the differential protein GLRX5, ferroptosis-related iron, GSH, MDA, SOD, ROS, liperfluo, and mitochondrial morphology were analyzed. Then, the proteins of GLRX5-mediated iron-starvation response and SLC7A11/GPX4 were analyzed. Finally, OGD/R-induced SH-SY5Y cells upon GLRX5 silencing were constructed to demonstrate that PCB-DS improved CIRI by GLRX5-mediated ferroptosis. PCB-DS better alleviated CIRI through decreasing neurological score, reducing the infarct volume, and suppressing the release of inflammatory cytokines than DS. Proteomics suggested that PCB-DS may ameliorate CIRI by inhibiting GLRX5-mediated ferroptosis, which was different from DS. PCB-DS reversed the abnormal mitochondrial morphology, iron, GSH, MDA, SOD, ROS, and liperfluo to inhibit ferroptosis in vitro and in vivo. PCB-DS directly activated GLRX5 suppressing the iron-starvation response and downregulated the SLC7A11/GPX4 signaling pathway to inhibit ferroptosis. Finally, silencing GLRX5 activated the iron-starvation response in SH-SY5Y cells and PCB-DS unimproved OGD/R injury upon GLRX5 silencing. Different from DS, PCB-DS suppressed ferroptosis to alleviate CIRI through inhibiting GLRX5-mediated iron-starvation response. These findings give a comprehensive understanding of the molecular mechanism underlying the effect of PCB-DS against CIRI and provide evidence to assess the product in clinical studies. [Display omitted] [ABSTRACT FROM AUTHOR]

Stroke is a complex physiological process associated with intestinal flora dysbiosis and metabolic disorders. Dan-deng-tong-nao capsule (DDTN) is a traditional Chinese medicine used clinically to treat cerebral ischemia-reperfusion injury (CIRI) for many years. However, little is known about the effects of DDTN in the treatment of CIRI from the perspective of gut microbiota and metabolites. This study aimed to investigate the regulatory roles of DDTN in endogenous metabolism and gut microbiota in CIRI rats, thus providing a basis for clinical rational drug use and discovering natural products with potential physiological activities in DDTN for the treatment of CIRI. The chemical composition of DDTN in vitro and in vivo was investigated using ultra-high performance liquid chromatography-high resolution mass spectrometry (UHPLC HRMS), followed by target prediction using reverse molecular docking. Secondly, a biological evaluation of DDTN ameliorating neural damage in CIRI was performed at the whole animal level. Then, an integrated omics approach based on UHPLC HRMS and 16S rRNA sequencing was proposed to reveal the anti-CIRI effect and possible mechanism of DDTN. Finally, exploring the intrinsic link between changes in metabolite profiles, changes in the intestinal flora, and targets of components to reveal DDTN for the treatment of CIRI. A total of 112 chemical components of DDTN were identified in vitro and 10 absorbed constituents in vivo. The efficacy of DDTN in the treatment of CIRI was confirmed by alleviating cerebral infarction and neurological deficits. After the DDTN intervention, 21 and 26 metabolites were significantly altered in plasma and fecal, respectively. Based on the fecal microbiome, a total of 36 genera were enriched among the different groups. Finally, the results of the network integration analysis showed that the 10 potential active ingredients of DDTN could mediate the differential expression of 24 metabolites and 6 gut microbes by targeting 25 target proteins. This study was the first to outline the landscapes of metabolites as well as gut microbiota regulated by DDTN in CIRI rats using multi-omics data, and comprehensively revealed the systematic relationships among ingredients, targets, metabolites, and gut microbiota, thus providing new perspectives on the mechanism of DDTN in the treatment of CIRI. [Display omitted] [ABSTRACT FROM AUTHOR]

Cerebral ischemia reperfusion (I/R) is a therapeutic strategy for ischemia; however, it usually causes injury by the aspect of inflammation and neuron apoptosis. This investigation aims to investigate the protective effects of phytic acid (IP6) for cerebral I/R injury in vitro. PC-12 cells under Oxygen and glucose deprivation/reperfusion (OGD/R) were performed to mimic cerebral I/R. IP6 was pretreated before PC-12 cells under OGD/R treatment. The data showed that IP6 activated the expression of sestrin2 in OGD/R injured PC-12 cells. IP6 inhibited OGD/R induced inflammation, oxidative stress, and apoptosis by activating sestrin2. Besides, p38 MAPK may mediate the effects of sestrin2 activated by IP6. Therefore, IP6 can be a potential drug to prevent neurological damage in cerebral I/R injury. Phytic acid exerts protective effects in cerebral I/R injury by activating sestrin2. [ABSTRACT FROM AUTHOR]

OBJECTIVE: To explore the specific mechanism of sevoflurane in alleviating cerebral ischemia-reperfusion injury (CIRI) in rats through the c-Jun N-terminal kinase (JNK) signaling pathway. MATERIALS AND METHODS: A total of 60 male specific pathogen-free Sprague-Dawley rats were randomly divided into sham group (n=20), model group (n=20), and sevoflurane group (n=20). In the sevoflurane group, sevoflurane (2.5%) was inhaled for 60 min at 24 h before the blockage of cerebral blood supply. The CIRI model was established using the suture method in the model group and sevoflurane group, while the right common carotid artery and external carotid artery were separated and ligated only, without suture placement, in the sham group. At 24 h after reperfusion, the neurological defi- cit score in each group was calculated, the water content in brain tissues in each group was detected based on dry-wet weight ratio, the infarction volume of brain tissues in each group was detected via 2,3,5-triphenyltetrazolium chloride (TTC) staining, and the apoptosis rate of brain cells in each group was detected using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. Moreover, the protein levels of JNK, p-JNK, B-cell lymphoma-2 (Bcl-2), and the Bcl-2 associated X protein (Bax) in brain tissues were determined using Western blotting, and the gene expressions of Bax and Bcl-2 in brain tissues were determined through fluorescence quantitative Polymerase Chain Reaction (qPCR). RESULTS: It was found that the water content in brain tissues and the cerebral infarction volume were significantly increased in the model group compared with those in the sham group (p<0.01, p<0.01), while they were notably decreased in the sevoflurane group compared with those in the model group (p<0.05, p<0.01). The neurological deficit score was significantly higher in the model group than that in the shsham group (p<0.01), while it was remarkably lower in the sevoflurane group than that in the model group (p<0.01). According to the results of the TUNEL assay, the model group had an evidently higher apoptosis rate of brain cells than the sham group (p<0.01), while the sevoflurane group had a lower apoptosis rate of brain cells than the model group (p<0.05). Besides, the results of Western blotting revealed that the model group exhibited remarkably increased protein levels of JNK, p-JNK, and Bax (p<0.05, p<0.01, p<0.01) and a remarkably decreased protein level of Bcl- 2 (p<0.01) compared with the sham group. Sevo- flurane group had decreased protein levels of JNK, p-JNK, and Bax (p<0.05, p<0.01, p<0.01) and an increased protein level of Bcl-2 (p<0.05) in comparison with the model group. In addition, the gene expression of Bcl-2 significantly declined (p<0.01), and that of Bax remarkably rose (p<0.01) in the model group compared with those in the sham group, while the contrary is the case in the sevoflurane group compared with those in the model group (p<0.05, p<0.01). CONCLUSIONS: Sevoflurane can regulate the protein and gene expressions of Bax and Bcl-2 and reduce apoptosis in CIRI by regulating the JNK signaling pathway, thereby exerting a protective effect on brain tissues and improving the symptoms of neurological deficit. [ABSTRACT FROM AUTHOR]

OBJECTIVE: This study aimed to explore the role of miR-155-5p in middle cerebral artery occlusion/reperfusion (MCAO/R) model in rats and oxygen-glucose deprivation/reoxygenation (OGD/R)-induced SH-SY5Y cells. In addition, this study also aimed to explore the underlying mechanisms to expect that miR-155- 5p may be investigated as a new and effective diagnostic and therapeutic target for ischemic stroke. MATERIALS AND METHODS: The in vivo MCAO/R rat model and in vitro OGD/R cell model were established. The miR-155-5p mRNA expression was detected by quantitative Reverse Transcription-Polymerase Chain Reaction (qRTPCR). Dual specificity ATPase (DUSP) 14 was predicted to be a potential target of miR-155-5p by TargetScan. The targeting relationship was confirmed by Luciferase assay. The cell viability was determined using the Cell Counting Kit- 8 (CCK-8). The expression level of inflammatory cytokines, including tumor necrosis factor alpha (TNF-a), interleukin-1ß (IL-1ß), and interleukin-6 (IL-6) levels were detected by Enzyme-Linked Immunosorbent Assay (ELISA). Western blot was used to detect the protein expression of DUSP14, the apoptotic protein Cleaved cysteine- aspartic acid protease (caspase)-3, and Cleaved PARP, as well as nuclear factor kappa B (NF-κB) and MAPKs signaling pathways related proteins. RESULTS: MiR-155-5p was upregulated in both MCAO/R rats and OGD/R-induced SHSY5Y cells. MiR-155-5p knockdown inhibited OGD/R-induced cell injury and inflammation, as well as MCAO/R-induced brain injury. MiR-155- 5p regulated the NF-κB and MAPKs signaling pathways by targeting DUSP14. DUSP14 knockdown partially reversed the protective effect of miR-155-5p knockdown on OGD/R-induced SHSY5Y cell injury and inflammation. CONCLUSIONS: MiR-155-5p accelerates cerebral I/R injury via targeting DUSP14 by regulating NF-κB and MAPKs signaling pathways. Inhibition of miR-155-5p significantly reduces apoptosis and brain injury. These results indicated that miR-155-5p plays a key role in cerebral I/R injury and has the potential to be explored as a new target for ischemic stroke. [ABSTRACT FROM AUTHOR]

[Display omitted] • MC-ELNs reduce mortality, protect nerve functions in delayed t-PA-treated I/R rats. • MC-ELNs alleviate delayed t-PA-induced hemorrhagic transformation and BBB injury. • MC-ELNs upregulate the expression of ZO-1 and claudin-5. • MC-ELNs decrease ONOO− level and inhibit ONOO− induced HMGB1 and MMP-9 expression. 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. [ABSTRACT FROM AUTHOR]

Cerebral ischemia has the characteristics of high incidence, mortality, and disability, which seriously damages people's health. Cerebral ischemia-reperfusion injury is the key pathological injury of this disease. However, there is a lack of drugs that can reduce cerebral ischemia-reperfusion injury in clinical practice. At present, a few studies have provided some evidence that nuciferine can reduce cerebral ischemia-reperfusion injury, but its specific mechanism of action is still unclear, and further research is still needed. In this study, PC12 cells and SD rats were used to construct OGD/R and MCAO/R models, respectively. Combined with bioinformatics methods and experimental verification methods, the purpose of this study was to conduct a systematic and comprehensive study on the effect and mechanism of nuciferine on reducing inflammation induced by cerebral ischemia-reperfusion injury. Nuciferine can improve the cell viability of PC12 cells induced by OGD/R, reduce apoptosis, and reduce the expression of inflammation-related proteins; it can also improve the cognitive and motor dysfunction of MCAO/R-induced rats by behavioral tests, reduce the area of cerebral infarction, reduce the release of inflammatory factors TNF-α and IL-6 in serum and the expression of inflammation-related proteins in brain tissue. Nuciferine can reduce the inflammatory level of cerebral ischemia-reperfusion injury in vivo and in vitro models by acting on the PI3K/Akt/NF-κB signaling pathway, and has the potential to be developed as a drug for the treatment of cerebral ischemia-reperfusion injury. [Display omitted] [ABSTRACT FROM AUTHOR]

Cerebral ischemia-reperfusion injury (CIRI) is a phenomenon that pathological injury of ischemic brain tissue is further aggravated after the restoration of blood supply. The complex pathological mechanism of CIRI has led to the failure of multiple neuroprotective agents in clinical studies. Salvianolic acid A (SAA) is a neuroprotective extract from Salvia miltiorrhiza Bge., with significant pharmacological activities in the treatment of brain injury. However, the neuroprotective mechanisms of SAA remain unclear. To explore the potential protective effect of SAA on CIRI and its mechanism, and to provide experimental basis for the research of new drugs for CIRI. A model of transient middle cerebral artery occlusion (tMCAO) in rats was used to simulate clinical CIRI, and the neuroprotective effect of SAA on tMCAO rats was investigated within 14 days after reperfusion. The improvement effects of SAA on cognitive impairment of tMCAO rats were investigated by behavioral tests from days 7–14. Finally, the neuroprotective mechanism of SAA was investigated on day 14. The neuroprotective effects and mechanism of SAA were investigated by behavioral tests, HE and TUNEL staining, RNA sequence (RNA-seq) analysis and Western blot in tMCAO rats. The brain protective effects of SAA were achieved by alleviating cerebral infarction, cerebral edema, cerebral atrophy and nerve injury in tMCAO rats. Meanwhile, SAA could effectively improve the cognitive impairment and pathological damage of hippocampal tissue, and inhibit cell apoptosis in tMCAO rats. Besides, SAA could provide neuroprotective effects by up-regulating the expression of Bcl-2, inhibiting the activation of Caspase 3, and regulating PKA/CREB/c-Fos signaling pathway. SAA can significantly improve brain injury and cognitive impairment in CIRI rats, and this neuroprotective effect may be achieved through the anti-apoptotic effect and the regulation of PKA/CREB/c-Fos signaling pathway. SAA promoted the expression of PKA, resulting in increased phosphorylation of CREB and decreased phosphorylation of c-Fos. In addition, SAA increased the ratio of Bcl-2/Bax and inhibited the expression of Cleaved Caspase 3, which inhibited the apoptosis of neurons. SAA provided neuroprotective effects and improved cognitive impairment caused by cerebral ischemia-reperfusion injury in tMACO rats by inhibiting neuronal apoptosis along with the regulation of PKA/CREB/c-Fos signaling pathway. [Display omitted] [ABSTRACT FROM AUTHOR]

Abstract Scavenging reactive oxygen species (ROS) by antioxidants is the important therapy to cerebral ischemia-reperfusion injury (CIRI) in stroke. The antioxidant with novel dual-antioxidant mechanism of directly scavenging ROS and indirectly through antioxidant pathway activation may be a promising CIRI therapeutic strategy. In our study, a series of chalcone analogues were designed and synthesized, and multiple potential chalcone analogues with dual antioxidant mechanisms were screened. Among these compounds, the most active 33 not only conferred cytoprotection of H 2 O 2 -induced oxidative damage in PC12 cells through scavenging free radicals directly and activating NRF2/ARE antioxidant pathway at the same time, but also played an important role against ischemia/reperfusion-related brain injury in animals. More importantly, in comparison with mono-antioxidant mechanism compounds, 33 exhibited higher cytoprotective and neuroprotective potential in vitro and in vivo. Overall, our findings showed compound 33 could emerge as a promising anti-ischemic stroke drug candidate and provided novel dual-antioxidant mechanism strategies and concepts for oxidative stress-related diseases treatment. Graphical abstract Chalcone analogue 33 conferred protection of PC12 cells against H 2 O 2 insult with novel dual-antioxidant mechanism of directly scavenging reactive oxygen species (ROS) and indirectly through antioxidant pathway activation, and the effect of 33 was more pronounced than that of mono-antioxidant mechanism compounds after MCAO and BCAO in animals. fx1 [ABSTRACT FROM AUTHOR]

Targeting neuronal functional restoration can be an efficacious regimen for treating neurological diseases, including cerebral ischemia-reperfusion injury (CIRI). Then, there is great significance for screening natural ingredient-played neuroprotection with therapeutical safety. Ginsenoside Re (Re), a nutraceutical molecule isolated from Panax ginseng , is previously proven to possess the neuroprotective benefit against CIRI. However, anti-CIRI regulatory mechanisms of Re action have not been totally explicated. In present study, we performed a network pharmacology approach accompanied with molecular docking analysis to illustrate the neuroprotective activity and biological mechanism of Re in the treatment of CIRI, characterized with bioinformatics findings by using gene ontology (GO) functional assay, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Data from integrated bioinformatics analysis exhibited that 60 candidate genes were identified as the therapeutic potentials of Re against CIRI related to autophagy. Most of these genes were found to be regulated functionally for CIRI after Re treatment. According to topological parameter analysis, total 30 core targets in Re against CIRI relating autophagy were discovered. The molecular pathways enriched from core targets might be implicated in neural regeneration, neural restoration and immunoregulation. Further computational assay identified 3 key proteins including protein kinase B (AKT1), phosphatidylinositol-4,5-bisphosphate 3-kinase, catalytic subunit alpha (PIK3CA) and heat shock protein 90 AA1 (HSP90AA1), in which these proteins' functions were related to the actions and pathways of autophagy. Current findings from this study combine network pharmacology approach and molecular docking analysis to uncover the core targets and molecular mechanisms of Re in the treatment of CIRI related to autophagy, and then we extrapolate that Ginsenoside Re may be used for the potential application for treating CIRI. [Display omitted] • All 60 candidate genes were identified as potential anti-CIRI targets of Re related to autophagy. • All core targets of Re against CIRI related to autophagy were identified. • Molecular mechanisms in Re against CIRI linked to neural regeneration, neural restoration and immunoregulation. • Computational assay identified key proteins including AKT1, PIK3CA and HSP90AA1. • These bioinformatics findings showed Re may be clinical application for treating CIRI. [ABSTRACT FROM AUTHOR]

OBJECTIVE: To investigate the changes of peroxisome proliferator-activated receptor gamma (PPAR γ) in focal cerebral ischemia- reperfusion injury, and to explore the effect and mechanism of mifepristone on the cerebral ischemia-reperfusion injury. MATERIALS AND METHODS: Male Sprague-Dawley (SD) rats were selected, and the middle cerebral artery occlusion and reperfusion (MCAO/R) rat model was constructed using the longa's suture-occluded method. The sham operation group was not inserted with occlusion sutures. All experimental rats were divided into four groups: the sham operation group (SHA group), the MCAO/R model group (MCR group), the mifepristone intervention group (MIF group) (3 mg/kg, intragastric administration), and the mifepristone + bisphenol A diglycidyl ether (BADGE) intervention group (MIF+BAD group) [3 mg/kg mifepristone (intragastric administration) + 30 mg/kg BADGE (intraperitoneal injection)]. The long's scoring method (5 grades) was applied for scoring after reperfusion, at the time when the animals woke up, and at 48 h after awaking before execution, respectively. 48 h after the model was successfully established, triphenyl tetrazolium chloride (TTC) staining was performed to calculate the volume of cerebral infarction, and Nissl staining was conducted to observe the cranial nerve tissue morphology. Meanwhile, immune-histochemical staining was used to detect PPAR γ. Moreover, the protein expression levels of PPAR γ, tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), interleukin-6 (IL-6), matrix metalloproteinase- 2 (MMP-2), MMP-9 and tissue inhibitor of metalloproteinase 1 (TIMP-1) were examined by Western blotting (WB). RESULTS: Mifepristone could significantly enhance the neurological function after cerebral ischemia-reperfusion injury, reduce the volume of cerebral infarction, and improve the morphology of nerve tissues in rats. The expression of PPAR γ in the brain tissues of rats after cerebral ischemia-reperfusion injury markedly declined, whereas mifepristone could remarkably increase the protein expression of PPAR γ. After mifepristone intervention, the protein levels of TNF-α, IL-1β, IL-6, MMP-2, and MMP-9 in the infarcted brain tissues of rats were markedly decreased, while the expression of the TIMP-1 protein was increased. When combined with BADGE, the effect of mifepristone was partially offset. CONCLUSIONS: Mifepristone acts as a PPAR γ agonist, and relieves cerebral ischemia-reperfusion injury by restoring the balance between MMPs and TIMPs and inhibiting inflammatory cytokines. [ABSTRACT FROM AUTHOR]

OBJECTIVE: The aim of the study was to investigate the anti-inflammatory effect of sevoflurane post-conditioning on cerebral ischemia- reperfusion injury in rats. MATERIALS AND METHODS: Thirty Sprague Dawley (SD) rats were randomly divided into 3 groups: sham operation group (Sham), ischemia/ reperfusion injury (I/R) group and sevoflurane post-conditioning group (Se). Hematoxylin-eosin (HE) staining was used to observe the inflammatory response in the brain tissue. The levels of TNF-α, IL- 1β, IL-6 in serum were measured by ELISA. The mRNA and protein expression of TLR4 and NF-κB p65 were detected by RT-PCR and Western blot in the brain tissue. RESULTS: The post-conditioning of sevoflurane decreased the level of inflammatory reaction in ischemic-reperfusion rat cerebral infarction area and reduced the levels of pro-inflammatory cytokines such as TNF-α, IL-1β, IL-6 in rats with ischemia- reperfusion injury. In addition, after treatment with sevoflurane, the mRNA and protein expression of TLR4 and NF-κBp65 in TLR4/NF-κB pathway was inhibited. CONCLUSIONS: Sevoflurane post-conditioning can decrease the inflammatory reaction in cerebral infarct area induced by cerebral ischemia- reperfusion injury in rats. The neuroprotective effect mechanism of sevoflurane may be related to TLR4/NF-κB signaling pathway. [ABSTRACT FROM AUTHOR]

• AIM2 is a target gene of miR-193b-5p. • Over expression of miR-193b-5p reduced pyroptosis and increased cell activity in OGD/R PC12 cells by AIM2 pathway. • miR-193b-5p-overexpressing exosomes had a greater effect in inhibiting AIM2 pathway-mediated pyroptosis in both in vivo and in vitro assays. • BMSC-Exos attenuate cerebral I/R injury by inhibiting AIM2 pathway-mediated pyroptosis through miR-193b-5p delivery. Ischemic stroke represents a major factor causing global morbidity and death. Bone marrow mesenchymal stem cell (BMSC)-derived exosomes (Exos) have important effects on treating ischemic stroke. Here, we investigated the therapeutic mechanism by which BMSC-derived exosomal miR-193b-5p affects ischemic stroke. luciferase assay was performed to evaluate the regulatory relationship of miR-193b-5p with absent in melanoma 2 (AIM2). Additionally, an oxygen-glucose deprivation/reperfusion (OGD/R) model was constructed for the in vitro assay, while a middle cerebral artery occlusion (MCAO) model was developed for the in vivo assay. After exosome therapy, lactate dehydrogenase and MTT assays were conducted to detect cytotoxicity and cell viability, while PCR, ELISA, western blotting assay, and immunofluorescence staining were performed to detect changes in the levels of pyroptosis-related molecules. TTC staining and TUNEL assays were performed to assess cerebral ischemia/reperfusion (I/R) injury. In the luciferase assay, miR-193b-5p showed direct binding to the 3ʹ-untranslated region of AIM2. In both in vivo and in vitro assays, the injected exosomes could access the sites of ischemic injury and could be internalized. In the in vitro assay, compared to normal BMSC-Exos, miR-193b-5p-overexpressing BMSC-Exos showed greater effects on increasing cell viability and attenuating cytotoxicity; AIM2, GSDMD-N, and cleaved caspase-1 levels; and IL-1β/IL-18 generation. In the in vivo assay, compared to normal BMSC-Exos, miR-193b-5p-overexpressing BMSC-Exos showed greater effects on decreasing the levels of these pyroptosis-related molecules and infarct volume. BMSC-Exos attenuate the cerebral I/R injury in vivo and in vitro by inhibiting AIM2 pathway-mediated pyroptosis through miR-193b-5p delivery. [ABSTRACT FROM AUTHOR]

Jasminoidin (JA) and ursodeoxycholic acid (UA) were shown to act synergistically against ischemic stroke (IS) in our previous studies. To investigate the holistic synergistic mechanism of JA and UA on cerebral ischemia. Middle cerebral artery obstruction reperfusion (MCAO/R) mice were used to evaluate the efficacy of JA, UA, and JA combined with UA (JU) using neurological function testing and infarct volume examination. High-throughput RNA-seq combined with computational prediction and function-integrated analysis was conducted to gain insight into the comprehensive mechanism of synergy. The core mechanism was validated using western blotting. JA and UA synergistically reduced cerebral infarct volume and alleviated neurological deficits and pathological changes in MCAO/R mice. A total of 1437, 396, 1080, and 987 differentially expressed genes were identified in the vehicle, JA, UA, and JU groups, respectively. A strong synergistic effect between JA and UA was predicted using chemical similarity analysis, target profile comparison, and semantic similarity analysis. As the 'long-tail' drugs, the top 20 gene ontology (GO) biological processes of JA, UA, and JU groups primarily reflected inflammatory response and regulation of cytokine production, with specific GO terms of JU revealing enhanced regulation on immune response and tumor necrosis factor superfamily cytokine production. Comparably, the Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling of common targets of JA, UA, and JU focused on extracellular matrix organization and signaling by interleukins, immune system, phagosomes, and lysosomes, which interlock and interweave to produce the synergistic effects of JU. The characteristic signaling pathway identified for JU highlighted the crosstalk between autophagy activation and inflammatory pathways, especially the Dectin-1-induced NF-κB activation pathway, which was validated by in vivo experiments. JA and UA can synergistically protect cerebral ischemia-reperfusion injury by attenuating Dectin-1-induced NF-κB activation. The strategy integrating high throughput data with computational models enables ever-finer mapping of 'long-tail' drugs to dynamic variations in condition-specific omics to clarify synergistic mechanisms. [Display omitted] [ABSTRACT FROM AUTHOR]

Stroke is a leading cause of long-term disability and death in the United States. Currently, tissue plasminogen activator (tPA), is the only Food and Drug Administration-approved treatment for acute ischemic stroke. However, the use of tPA is restricted to a small subset of acute stroke patients due to its limited 3-h therapeutic time window. Given the limited therapeutic options at present and the multi-factorial progression of ischemic stroke, emphasis has been placed on the discovery and use of combination therapies aimed at various molecular targets contributing to ischemic cell death. Protein kinase C (PKC) and Akt (protein kinase B) are serine/threonine kinases that play a critical role in mediating ischemic-reperfusion injury and cellular growth and survival, respectively. The present review will examine the role of PKC and Akt in the cellular response to ischemic-reperfusion injury. [ABSTRACT FROM AUTHOR]

Background The pathogenesis of cerebral ischemia-reperfusion injury (CIRI) is not completely clear and therapies are limited now. Therefore, our study aimed to investigate the possible pathogenesis and preventive approach of CIRI through analyzing changes of aspartate (Asp), glutamate (Glu), mitochondrial calcium (MCa), calmodulin (CaM), and malondialdehyde (MDA) contents and ultramicropathology in hippocampus and cerebral cortex of ischemic susceptible injured regions and the effect of monosialotetrahexosylganglioside (GM1) in the rat model of CIRI. Methods Contents of Asp, Glu, MCa, CaM, and MDA in hippocampus and cerebral cortex tissues were measured by a high-performance liquid chromatography, atomic absorption spectrophotometer, and ordinary spectrophotometer, respectively, changes of ultramicrostructure in neurons of the hippocampus CA1 region and frontal cerebral cortex were observed by a transmission electron microscope. Results Contents of Asp and Glu in hippocampus and cerebral cortex tissues of CIRI groups significantly decreased and contents of MCa, CaM, and MDA significantly increased than those in control groups, and the ultramicrostructure in neurons of the hippocampus CA1 region and frontal cerebral cortex revealed a significant damaged change, and GM1 significantly ameliorated changes of Asp, Glu, MCa, CaM, and MDA contents in hippocampus and ultrastructural changes in neurons of the hippocampus CA1 region and frontal cerebral cortex. Conclusions Our findings further support that the abnormal release and/or reuptake of excitatory amino acid neurotransmitters, the disordered calcium homeostasis and the excessive production and/or reduced elimination of reactive oxygen species contribute to the pathogenesis of CIRI, and GM1 can partially prevent these pathogenesises to exert the protective effect on CIRI. [ABSTRACT FROM AUTHOR]

Background: With the rising age of the global population, the incidence rate of cardiovascular and cerebrovascular diseases (CCVDs) is increasing, which causes serious public health burden. The efforts for new therapeutic approaches are still being sought since the treatment effects of existing therapies are not quite satisfactory. Chinese traditional medicine proved to be very efficient in the treatment of CCVDs. Well described and established in Chinese medicine, Astragali Radix, has been commonly administered in the prophylaxis and cure of CCVDs for thousands of years.Purpose: This review summarized the action mode and mechanisms of Astragali Radix phytochemicals on CCVDs, hoping to provide valuable information for the future application, development and improvement of Astragali Radix as well as CCVDs treatment.Methods: A plenty of literature on biological active ingredients of Astragali Radix used for CCVDs treatment were retrieved from online electronic PubMed and Web of Science databases.Results: This review highlighted the effects of five main active components in Astragali Radix including astragaloside Ⅳ, cycloastragenol, astragalus polysaccharide, calycosin-7-O-β-d-glucoside, and calycosin on CCVDs. The mechanisms mainly involved anti-oxidative damage, anti-inflammatory, and antiapoptotic through signaling pathways such as PI3K/Akt, Nrf2/HO-1, and TLR4/NF-κB pathway. In addition, the majority active constituents in AR have no obvious toxic side effects.Conclusion: The main active components of Astragali Radix, especially AS-IV, have been extensively summarized. It has been proved that Astragali Radix has obvious therapeutic effects on various CCVDs, including myocardial and cerebral ischemia, hypertension, atherosclerosis, cardiac hypertrophy, chronic heart failure. CAG possesses anti-ischemia activity without toxicity, indicating a worthy of further development. However, high-quality clinical and pharmacokinetic studies are required to validate the current studies. [ABSTRACT FROM AUTHOR]

The article presents a study which explores the neuroprotective effects and mechanisms of penehyclidine hydrochloride on cerebral ischemia-reperfusion injury in rats. The penehyclidine hydrochloride effect on N-methyl-D-aspartate receptors in neurons in hippocampus after oxygen-glucose deprivation was also examined. The findings show that penehyclidine hydrochloride could attenuate oxidative stress injury and neuronal apoptosis following cerebral ischemia-reperfusion.

The article presents a study which investigates the mature nerve growth factor (NGF) expression and nerve growth factor precursor (proNGF) in non-ischemic and ischemic cortices after cerebral ischemia-reperfusion injury. The study uses Wistar rats to make the experiment, wherein they were raised in quiet animal housing for one week without exposure to strong light. Results show that proNGF was the major form of NGF during the pathological process of cerebral ischemia-reperfusion injury.

Objective: Electroacupuncture (EA) pretreatment has been shown to alleviate cerebral ischemia-reperfusion (I/R) injury; however, the underlying mechanism remains unclear. To investigate the involvement of mTOR signaling in the protective role of EA in I/R-induced brain damage and mitochondrial injury.Methods: Sprague-Dawley male rats were pretreated with vehicle, EA (at Baihui and Shuigou acupoints), or rapamycin + EA for 30 min daily for 5 consecutive days, followed by the middle cerebral artery occlusion to induce I/R injury. The neurological functions of the rats were assessed using the Longa neurological deficit scores. The rats were sacrificed immediately after neurological function assessment. The brains were obtained for the measurements of cerebral infarct area. The mitochondrial structural alterations were observed under transmission electron microscopy. The mitochondrial membrane potential changes were detected by JC-1 staining. The alterations in autophagy-related protein expression were examined using Western blot analysis.Results: Compared with untreated I/R rats, EA-pretreated rats exhibited significantly decreased neurological deficit scores and cerebral infarct volumes. EA pretreatment also reversed I/R-induced mitochondrial structural abnormalities and loss of mitochondrial membrane potential. Furthermore, EA pretreatment downregulated the protein expression of LC3-II, p-ULK1, and FUNDC1 while upregulating the protein expression of p-mTORC1 and LC3-I. Rapamycin effectively blocked the above-mentioned effects of EA.Conclusion: EA pretreatment at Baihui and Shuigou alleviates cerebral I/R injury and mitochondrial impairment in rats through activating the mTORC1 signaling. The suppression of autophagy-related p-ULK1/FUNDC1 pathway is involved in the neuroprotective effects of EA. [ABSTRACT FROM AUTHOR]

Abstract: Objective: To explore the effects of Qingkailing injection (QKL) on transforming growth factor-β1 (TGF-β1) expression in the rats with cerebral ischemia-reperfusion injury. Methods: Healthy Wistar rats were randomly divided into 3 groups: normal group, model group, and QKL-treated group. Model of cerebral ischemia-reperfusion injury was copied by repeated occlusion of common carotid arteries (CCA) on both sides. The expression of glial fibrillary acidic protein (GFAP) and TGF-β1 in brain tissue was observed at different time intervals, using immunity histochemistry staining method. Results: At different time intervals after cerebral ischemia-reperfusion injury, the expression of GFAP and TGF-β1 in brain tissue increased by different degrees, whereas QKL decreased them by different degrees. Conclusion: QKL could probably reduce the expression of TGF-β1 by restraining the activity of astrocytes (AST). [Copyright &y& Elsevier]

Background: Thymus quinquecostatus Celak. has been widely used as a spice and a folk medicine for relieving exterior syndrome and alleviating pain in China.Purpose: To explore the protective effects and the underlying mechanism against cerebral ischemia-reperfusion injury (CIRI) of the T. quinquecostatus combining with its chemical composition.Study Design and Methods: High-polar extract (HPE) was extracted from T. quinquecostatus and polyphenols in HPE were enriched to obtain polyphenol-rich fraction (PRF) using Macroporous resin. The free radicals and zebrafish embryos were used to compare the antioxidant activities of HPE and PRF in vitro and in vivo. Then, the transient middle cerebral artery occlusion (tMCAO) model was established in rats. Neurological deficit score, infarction rate, morphology and apoptosis of neurons were examined to investigate the protective effects of PRF on CIRI. The mRNA and protein levels of nuclear factor erythroid 2-related factor 2 (Nrf2) and hemeoxygenase-1 (HO-1) and the activities of downstream antioxidant enzymes in ischemia tissues were determined to clarify the underlying mechanisms. Also, reactive oxygen species (ROS) level in zebrafish embryos were detected after incubation with PRF for a short time (2 h) to investigate whether PRF could directly eliminate free radicals. Finally, chemical composition of PRF were analyzed to investigate the material basis for antioxidant activity and anti-CIRI effect.Results: Compared with HPE, PRF showed stronger antioxidant activities. PRF exhibited obvious protective effects including ameliorating neurological deficit, lowering infarction rate, and improving the cellular morphology in hippocampus CA1 and cortex after tMCAO. TUNEL staining suggested PRF dose-dependently improved the apoptosis of the neurons in ischemic cortex. RT-qPCR and Western Blot results suggested that PRF regulated oxidative stress (OS) via activating the Keap1/Nrf2/HO-1 signaling pathway. Also, PRF could directly scavenge excessive ROS in zebrafish embryos after a short-time PRF incubation. The anti-CIRI effect might be primarily attributed to the abundant polyphenols in PRF, including flavonoids, polymethoxylated flavonoids, flavonoid glycosides, and phenolic acids.Conclusion: T. quinquecostatus contains abundant polyphenols and exhibited a good protective effect against CIRI via dual antioxidant mechanisms, providing a reference for further research and application for this plant. [ABSTRACT FROM AUTHOR]

Background: To investigate the value of plasma high mobility group box protein 1 (HMGB1) in evaluating the prognosis of cerebral ischemia-reperfusion injury (CIRI) in ischemic stroke patients.Methods: 132 ischemic stroke patients were recruited. Before and after thrombolytic therapy at 2 h, 6 h, 12 h, 24 h, and 36 h, the Glasgow Coma Scale (GCS) and National Institutes of Health Stroke Scale (NIHSS) were recorded. The Modified Rankin scale (mRS) was used to assess the prognosis at 3 months.Results: The NIHSS score, GCS score and plasma HMGB1 level peaked at 6 h after thrombolytic therapy, and plasma HMGB1 level was positively correlated with infarct volume and NIHSS score, and negatively correlated with GCS score. Plasma HMGB1 level at 6 h had the highest value in identifying patients with poor unfavorable functional outcome after 3 months, with a sensitivity of 86.8% and a specificity of 74.0%. Logistic regression results showed that plasma HMGB1 had a strong association with unfavorable functional outcome [odds ratio (OR) =1.621, P<0.001]. After adjusting for infarct volume and NIHSS score did not attenuate the association (OR=1.381, P=0.005). Finally, we found that plasma HMGB1 at 6 h had the highest value in identifying patients with non-survival after 3 months (χ2=28.655, P<0.001). Logistic regression results showed that plasma HMGB1 had a strong association with non-survival (OR=2.315, P<0.001). After adjusting for infarct volume and NIHSS score did not attenuate the association (OR=2.013, P<0.001).Conclusion: Plasma HMGB1 exerts a good predictive value for CIRI in ischemic stroke patients, and its increased expression is correlated with worse prognosis. [ABSTRACT FROM AUTHOR]