Cerebral Infarction Area Research Articles

Tetramethylpyrazine Attenuates Cerebral Ischemia-Reperfusion Injury by Inhibiting Ferroptosis via the AMPK / Nrf2 Pathways.

Ferroptosis is involved in the development and exacerbation of cerebral ischemia-reperfusion injury (CIRI), and its inhibition can alleviate CIRI. Tetramethylpyrazine (TMP) is used for the treatment of ischemic stroke. However, the mechanism by which TMP regulates ferroptosis in CIRI is yet to be explored. This study demonstrated the effects of TMP on ferroptosis and CIRI, including the roles of the adenosine 5'-monophosphate-activated protein kinase (AMPK)/nuclear factor erythroid-2-related factor 2 (Nrf2) signaling pathway. A Sprague-Dawley rat middle cerebral artery occlusion/reperfusion (MCAO/R) model was generated. The extent of neuronal injury was measured using 2,3,5-triphenyl tetrazolium chloride staining and Garcia neurological scoring and behavior was evaluated using open-field tests. Ferroptosis-related indexes were examined and ferroptosis-related proteins were detected using western blotting. The binding modes of TMP and AMPK were evaluated using molecular docking and molecular dynamics simulations. MCAO/R rats showed a reduced cerebral infarct area and improved neurological function after TMP intervention. TMP reduced levels of Fe2+, 4-hydroxynonenal, malonaldehyde, and acyl-coenzyme synthetase long-chain family member 4 and increased levels of glutathione and glutathione peroxidase 4. Increased AMPK phosphorylation and Nrf2 expression were also detected. TMP bound tightly to the AMPKα subunit in silico, and the LEU157, VAL41, LEU33, VAL107, and TYR106 residues were important for binding. Our results indicate that TMP can alleviate CIRI by inhibiting ferroptosis via the activation of the AMPK/Nrf2 pathway, providing a theoretical basis for the clinical use of TMP in treating CIRI.

Traditional Chinese Medicine Borneol-Based Polymeric Micelles Intracerebral Drug Delivery System for Precisely Pathogenesis-Adaptive Treatment of Ischemic Stroke.

The scarcity of effective neuroprotective agents and the presence of blood-brain barrier (BBB)-mediated extremely inefficient intracerebral drug delivery are predominant obstacles to the treatment of cerebral ischemic stroke (CIS). Herein, ROS-responsive borneol-based amphiphilic polymeric NPs are constructed by using traditional Chinese medicine borneol as functional blocks that served as surface brain-targeting ligand, inner hydrophobic core for efficient drug loading of membrane-permeable calcium chelator BAPTA-AM, and neuroprotective structural component. In MCAO mice, the nanoformulation (polymer: 3.2 mg·kg-1, BAPTA-AM: 400 µg·kg-1) reversibly opened the BBB and achieved high brain biodistribution up to 12.7%ID/g of the total administered dose after 3 h post single injection, effectively restoring intracellular Ca2+ and redox homeostasis, improving cerebral histopathology, and inhibiting mitochondrial PI3K/Akt/Bcl-2/Bax/Cyto-C/Caspase-3,9 apoptosis pathway for rescuing dying neurons (reduced apoptosis cell from 59.5% to 7.9%). It also remodeled the inflammatory microenvironment in cerebral ischemic penumbra by inhibiting astrocyte over-activation, reprogramming microglia polarization toward an anti-inflammatory phenotype, and blocking NF-κB/TNF-α/IL-6 signaling pathways. These interventions eventually reduced the cerebral infarction area by 96.3%, significantly improved neurological function, and restored blood flow reperfusion from 66.2% to ≈100%, all while facilitating BBB repair and avoiding brain edema. This provides a potentially effective multiple-stage sequential treatment strategy for clinical CIS.

Na+-K+-ATPase/GLT-1 interaction participates in EGCG protection against cerebral ischemia-reperfusion injury in rats.

In China, stroke is the primary cause of adult death and disability. Because of the increased rate of blood vessel reperfusion, it is important to prevent cerebral ischemia-reperfusion injury, in which glutamate (Glu) excitotoxicity plays a critical role. The most important Glu transporter, GLT-1, is essential for the regulation of Glu, which is dependent on Na+-K+-ATPase (NKA)-induced ion concentration gradient differences. EGCG, a substance found in tea polyphenols, can reduce infarct areas in ischemia-reperfusion models, reduce stroke incidence, and prolong life in which NKA is involved. In this study, we investigated the potential of EGCG in protecting against cerebral ischemia-reperfusion injury by regulating the interaction between NKA and GLT-1. This study was designed to investigate the protective effects of EGCG against cerebral ischemia-reperfusion injury by modulating the interaction between NKA and GLT-1, utilizing both the rat middle cerebral artery occlusion/reperfusion (MCAO/R) model and the oxygen-glucose deprivation/reoxygenation (OGD/R) model in co-cultures of rat hippocampal neurons and astrocytes. The neuronal survival rate was assessed using CCK8, and the cerebral infarction area and neurological function were determined by TTC staining and neurological deficit scores. NKA activity was measured using an inorganic phosphorous detection method, and NKA and GLT-1 expression was detected using western blotting. The interaction between NKAα2 and GLT-1 was identified by co-immunoprecipitation (CoIP) assay, laser confocal microscopy, and Imaris 3D confocal rendering technology. An adenovirus vector with overexpression of NKAα2 was constructed, packaged, and injected into the rat lateral ventricle. Neurological function and the cerebral infarction area were identified, and the interaction between NKAα2 and GLT-1 was identified using CoIP assay. EGCG reduced the infarction area and neurological deficit scores, restored NKA activity, alleviated the decrease in membrane NKAα2 and GLT-1 expression, and relieved the uncoupling of NKAα2 and GLT-1 in the hippocampal CA1 after rat MCAO/R injury. By promoting the coupling of NKAα2 and GLT-1 in rat MCAO/R models, overexpression of NKAα2 reduced the cerebral infarction area and neurological impairment scores. EGCG improved cerebral ischemia-reperfusion injury by restoring NKA activity and increasing membrane GLT-1 expression due to NKA-GLT-1 interaction. For the first time, our findings demonstrate the critical role that NKA and GLT-1 colocalization plays in cerebral ischemia-reperfusion damage. Our findings provide new strategic directions for the pathogenesis and prevention of thrombolytic injury in the clinical treatment of stroke, while also serving as a basis for further development and utilization of EGCG.

Elemene mitigates oxidative stress and neuronal apoptosis induced by cerebral ischemia-reperfusion injury through the regulation of glutathione metabolism

Ethnopharmacological relevanceChinese materia medica (CMM) has a long history and extensive experience in treating ischemic stroke. Wen Ezhu, the rhizome of Curcuma wenyujin Y.H. Chen et C. Ling, is renowned for promoting blood circulation, dispersing blood stasis, alleviating pain, and eliminating masses. Promoting blood circulation and removing blood stasis are essential principles in Traditional Chinese Medicine for treating stroke. Consequently, Wen Ezhu is frequently used in clinical practice as a key CMM for treating stroke. The Elemene active fraction (ELE), a sesquiterpene compound extracted from Wen Ezhu, primarily consists of β-Elemene. It also contains β-Caryophyllene, γ-Elemene, and δ-Elemene isomers. ELE has shown potential pharmacological effects in various diseases, including ischemic stroke. However, its precise mechanism of action in treating stroke remains to be confirmed. Aim of the studyTo explore the therapeutic potential of ELE in acute ischemic stroke and elucidate its underlying mechanisms. Materials and methodsA rat model of middle cerebral artery occlusion reperfusion (MCAO/R) was used to evaluate ELE’s effects. Therapeutic efficacy was assessed through mNSS scoring, magnetic resonance imaging (MRI), tetrazolium chloride (TTC) staining, Hematoxylin and eosin (H&E), and Nissl staining. Non-targeted metabolomics identified key pathways, confirmed using biochemical analysis, immunohistochemistry, and Western blotting. ROS levels and apoptosis-related proteins were also evaluated. ResultsOur findings show that ELE administration significantly reduced the cerebral infarct area and lowered modified neurological severity scores (mNSS) in animals, indicating a strong neuroprotective effect. Metabolomics results highlight the glutathione (GSH) metabolic pathway as a key mechanism through which ELE exerts its therapeutic effects. Specifically, ELE upregulates glutathione reductase (GR) protein expression and downregulates glutathione peroxidase (GPX) expression. The regulatory process of ELE decreases oxidized glutathione (GSSG) levels and increases GSH levels, effectively reducing oxidative stress damage (lower reactive oxygen species levels) during CI/RI. This results in the downregulation of the pro-apoptotic protein Bax and the upregulation of the pro-survival protein Bcl-2, thus reducing neuronal apoptosis. ConclusionsELE protects neurons in MCAO/R rats through the GSH metabolism pathway, balancing GSH and GSSG levels to mitigate oxidative stress and enhance neuroprotection in cerebral ischemia/reperfusion injury.

Effect of electroacupuncture on neurological function in rats with cerebral ischemia-reperfusion injury based on AMPK/mTOR/ULK1 signaling pathway

To investigate the effect of electroacupuncture (EA) on neurological function in rats with cerebral ischemia-reperfusion injury (CIRI) by regulating adenosine monophosphate-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR)/Unc-51-like kinase 1 (ULK1) signaling pathway. Thirty-three male SD rats were randomly divided into a sham-operation group, a model group and an EA group, with 11 rats in each group. The right middle cerebral artery occlusion/reperfusion (MCAO/R) model was prepared by thread occlusion method in the model group and the EA group. In the sham-operation group, no thread was inserted after vascular separation. After the success of modeling, in the EA group, EA was applied to "Baihui" (GV 20) and "Zusanli" (ST 36) on the affected side, with disperse-dense wave, the frequency of 2 Hz/15 Hz, for 20 min, once a day. EA was delivered continuously for 3 days. On day 1 and day 3 of operation, the score of the modified neurological deficit scale (mNSS) was evaluated. After intervention completion, the cerebral infarction area was measured by the thiazolyl blue tetrazolium chloride (TTC) method. Nissl staining was used to observe the damage of cortical neurons on the ischemic side in each group. Using transmission electron microscopy, the ultrastructure of cortical neurons on the ischemic side was observed. With the immunofluorescence method adopted, the positive expression of the related protein 1 light chain 3 (LC3) and benzyl chloroform (Beclin-1) on the ischemic side was detected. The protein expression of p-AMPK, AMPK, p-mTOR, mTOR, pS757-ULK1, ULK1 and chelating ligand 1 (p62) in the ischemic cortex was detected using Western blot method. ① Compared with the sham-operation group, in the model group, the mNSS score increased (P<0.01), the percentage of infarction area was increased (P<0.01); the cortical neurons on the ischemic side were loosely distributed, with karyopyknosis and vacuolization, and the number of neurons was reduced (P<0.01); the cells were swollen and ruptured, mitochondrial shrunk, electron density higher, and there were a large number of autophagic debris. The positive expression of LC3 and Beclin-1 was elevated (P<0.01), and p-mTOR/mTOR, pS757-ULK1/ULK1 and the protein expression of p62 dropped (P<0.01) in the ischemic cortex. ② Compared with the model group, in the EA group, the mNSS score was reduced (P<0.05). The percentage of cerebral infarction area was descreased (P<0.01); and the neurons were regularly distributed, the number of neurons increased (P<0.01), the structure of mitochondria was clearer, the crest fracture alleviated, and the damage of neurons attenuated. The positive expression of LC3 and Beclin-1 was dropped (P<0.01), and p-AMPK/AMPK reduced (P<0.05), and p-mTOR/ mTOR, pS757-ULK1/ULK1 and the protein expression of p62 increased (P<0.01) in the ischemic cortex. EA at "Baihui" (GV 20) and "Zusanli" (ST 36) inhibits autophay, attenuates neurological deficit and cerebral pathological damage in CIRI rats to protect the nerves, which may be obtained by regulating AMPK/mTOR/ULK1 signaling pathway.

Study on protective effect and mechanism of Shouhui Tongbian Capsules on cerebral ischemia-reperfusion rat models based on metabolomics

This paper explored the protective effect and potential mechanism of Shouhui Tongbian Capsules(SHTB) on cerebral ischemia-reperfusion rat models. Rats were randomly divided into sham surgery group, model group, low-dose SHTB group(0.2 g·kg~(-1)·d~(-1)), high-dose SHTB group(SHTB g·kg~(-1)·d~(-1)), and an edaravone positive drug group(5.4 mg·kg~(-1)·d~(-1)), with 12 rats in each group. Rats were given continuous intragastric administration seven days before surgery, and the suture method was used to establish the cerebral ischemia-reperfusion injury(CIRI) rat model. Zea-Longa rating scale for neurological functions was used to assess the degree of neurological function impairment in rats; hematoxylin-eosin(HE) staining was used to observe the pathological damage of rats' brain tissue; TTC staining was used to measure the area of cerebral infarction in rats; enzyme-linked immunosorbent assay(ELISA) was used to detect the serum levels of interleukin-6(IL-6), interleukin-1β(IL-1β), and tumor necrosis factor-α(TNF-α) in each group. Western blot was used to detect the level of tight junction protein associated with the blood-brain barrier and intestinal barrier, as well as the protein expression of the TLR4/MyD88/NF-κB pathway in brain tissue. Changes in rats' brain tissue and metabolites in serum were detected by ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS), so as to explore the potential mechanism of SHTB treatment for CIRI in rats. Compared with the model group, SHTB could significantly alleviate the pathological damage to the brain of CIRI rats, reduce the volume of cerebral infarction, and lower the level of inflammation in the serum; Western blot results showed that SHTB could regulate the expression of tight junction proteins related to the blood-brain barrier and intestinal barrier in CIRI rats and downregulate the expression of TLR4, NF-κB, and MyD88 proteins in brain tissue. The UPLC-MS/MS results indicated that SHTB could significantly regulate the content of potential differential metabolites such as fatty acids, and serum and brain tissue are involved in pathways such as unsaturated fatty acid biosynthesis. SHTB could repair intestinal barrier function, reduce inflammation levels in the body, and improve the damaged blood-brain barrier, exerting a protective effect on brain nerves. Its mechanism may be achieved by balancing fatty acid metabolism and regulating the expression of TLR4/MyD88/NF-κB signaling pathway proteins.

Protective effects of puerarin combined with bone marrow mesenchymal stem cells on nerve injury in rats with ischemic stroke

ABSTRACT Background Bone marrow mesenchymal stem cells (BM-MSCs) transplantation shows promise for treating ischemic stroke, but the ischemic environment that follows cerebral infarction hinders the survival of transplanted cells. We aimed to study the effects of puerarin (Pue) in combination with BM-MSCs on cerebral ischemic injury. Methods After middle cerebral artery occlusion (MCAO) models were prepared by suture-occluded method, rats were randomly allocated to the sham, MCAO, Pue (50 mg/kg), BM-MSCs (2×106), and BM-MSCs+Pue groups. The neurological function, infarct area, levels of inflammation-related factors, brain tissue damage, apoptosis, BrdU, Beclin1, and LC3 levels were then assessed. Results Pue and BM-MSCs reduced the modified neurological severity score, cerebral infarction area, and serum inflammation-related factor levels for MCAO rats. Furthermore, Pue and BM-MSCs interventions ameliorated brain tissue damage, and repressed apoptosis of brain tissues in MCAO rats. Moreover, Pue or BM-MSCs enhanced BrdU expression, restrained LC3II/LC3I ratio and Beclin 1 expression in MCAO rats’ brain tissues. Importantly, the combination of Pue and BM-MSCs exhibited more pronounced effects on aforementioned outcomes. Conclusion The combination of Pue and BM-MSCs facilitated the recovery of neurological function in rats after cerebral ischemic damage, and the mechanisms may correlate with the repression of neuronal apoptosis, inflammation, and autophagy.

Management of large or giant Extracranial carotid artery aneurysms: a single-center experience

BackgroundExtracranial carotid artery aneurysm (ECCA) is an infrequent disease with an incidence of less than 1%. However, our understanding is still incomplete, and the preferred method to treat ECAA remains unknown.MethodsTo share our initial experience with treatment options for large ECCAs. We have retrospectively included 15 patients who underwent treatment at our institution from 2015 to 2022. The treatment modality, patient demography, aneurysm morphology, and clinical and radiographic follow-data were collected and analyzed in all patients.ResultsDuring the study period, 15 patients (with 19 ECCAs) were diagnosed and treated, of whom 8 (53.3%) were male. The average age of the patients was 53.6 years. The primary presenting symptoms was pulsatile neck mass (10/15, 66.7%). The etiology of ECAAs included atherosclerotic (6/15, 40.0%), infectious (3/15, 20.0%), and dissecting (1/15, 6.7%), and iatrogenic due to acupuncture (1/15, 6.7%). The mean ± SD maximal diameter of the aneurysms was 23.8 ± 14.1 mm, with more than half of patients having aneurysms larger than 25 mm (52.6%). 79.0% aneurysm had intraluminal thrombus at admission. Six patients underwent successful neurosurgical resection for a total of seven ECCAs. Five patients received endovascular interventional treatment. The remaining four patients who presented with seven ECCAs were placed under observation. The mean follow-up period was 28.1 months. Out of the patients who received treatment, it was discovered that 10 of them had completely occluded at the latest imaging study. During the course of conservative observation, it was observed that in one patient, the aneurysm disappeared and the parent vessel became thinner following anti-infection treatment. Out of all the patients, only one who was treated with Willis stent experienced a large area of cerebral infarction after treatment, which ultimately resulted in their death.ConclusionsECCA is rare and mostly present with mass effect. Neurosurgical treatment was more frequently feasible in large ECCAs, and endovascular surgery was the first choice for pseudoaneurysms and dissecting aneurysms. Anti-inflammation treatment was available for some infectious cases.

Effects of Motherwort on Biological Behavior of Brain Tissue in Rats with Ischemic Stroke and its Relationship with Phosphatidylinositol 3-Kinase/Protein Kinase B Signaling Pathway

Background Motherwort has a protective effect on the nervous system and can improve cerebral ischemia–reperfusion injury to a certain extent. However, the effect of motherwort on the biological behavior of the brain tissue of rats with ischemic stroke and its relationship with the phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) signaling pathway is still unclear. Purpose Therefore, we will explore the effect of motherwort on the biological behavior of the brain tissue of rats with ischemic stroke and its relationship with the PI3K/AKT signaling pathway. Methods There were 17 model rats in each group, divided into a control group, a model group, and a motherwort (low, medium, and high) dose group. Detect cell proliferation, invasion, and so on, and observe the expression of proteins and genes such as nuclear factor kappa B (NF-κB), AKT, Pl3K, and β-actin. Results After successfully constructing a rat model of ischemic stroke, motherwort can improve neurological damage and cerebral infarction in ischemic stroke and is closely related to PI3K/AKT signaling; in addition, motherwort can upregulate PI3K/AKT signaling and improve neurological damage and cerebral infarct size in ischemic stroke rats to a certain extent, and play a role in protecting neurons. Conclusion The antioxidant and antineuronal apoptosis effects of motherwort can improve neurological damage and cerebral infarction area of ischemic stroke rats by upregulating the expression level of the PI3K/AKT signaling pathway, thereby inhibiting nerve cells. Apoptosis further protects neurons and plays a role in slowing down disease progression.

Electroacupuncture alleviated post-stroke cognitive impairment via the mTOR/NLRP3-mediated autophagy-inflammatory pathway

BackgroundPost-stroke cognitive impairment (PSCI) severely reduces quality of life of patients with stroke. This study aimed to assess the effects of electroacupuncture (EA) on PSCI and the role of the mTOR/NLRP3-mediated autophagy-inflammatory pathway in this process.MethodsThe rat focal cerebral ischemia model was established using middle cerebral artery occlusion (MCAO). Following successful induction of the model, EA was applied to the bilateral Fengchi, Fengfu, and Dazhui acupoints, and brain tissue samples were collected on day 15. Cognitive function was assessed using the Morris water maze test. Cerebral infarct volume was quantified by Triphenyltetrazolium chloride (TTC) staining. Hematoxylin–eosin and TUNEL staining were performed to evaluate pathological changes and apoptosis rates. Apoptosis-, inflammation-, and autophagy-related biomarkers were measured, and autophagosomes were visualized using transmission electron microscopy.ResultsMCAO rats exhibited slower weight gain, reduced mobility, increased infarct size, pathological damage, and apoptosis, confirming successful establishment of the MCAO rat model. Following EA treatment, MCAO rats displayed faster weight gain, improved mobility, and shorter escape latency. EA also reduced the area of cerebral infarction and alleviated pathological damage and apoptosis in MCAO rats. Furthermore, EA downregulated IL-1β, IL-18, NLRP3, and LC3 II/LC3 I expression and upregulated p62, mTOR, and Beclin-1 expression in MCAO rats. EA treatment also decreased the number of autophagosomes in these rats.ConclusionsEA effectively mitigates post-stroke cognitive impairment by reducing apoptosis, inflammation, and autophagy through the regulation of the mTOR/NLRP3-mediated autophagy-inflammatory pathway, offering valuable therapeutic insights for stroke rehabilitation.

LncRNA Taurine Up-Regulated 1 Knockout Provides Neuroprotection in Ischemic Stroke Rats by Inhibiting Nuclear-Cytoplasmic Shuttling of HuR.

Background: Long non-coding RNA taurine-upregulated gene 1 (TUG1) is involved in various cellular processes, but its role in cerebral ischemia-reperfusion injury remains unclear. This study investigated TUG1's role in regulating the nucleocytoplasmic shuttling of human antigen R (HuR), a key apoptosis regulator under ischemic conditions. Methods: CRISPR-Cas9 technology was used to generate TUG1 knockout Sprague Dawley rats to assess TUG1's impact on ischemic injury. The infarct area and neuronal apoptosis were evaluated using TUNEL, hematoxylin and eosin (HE), and TTC staining, while behavioral functions were assessed. Immunofluorescence staining with confocal microscopy was employed to examine TUG1-mediated HuR translocation and expression changes in the apoptosis-related proteins COX-2 and Bax. Results: TUG1 knockout rats showed significantly reduced cerebral infarct areas, decreased neuronal apoptosis, and improved neurological functions compared to controls. Immunofluorescence staining revealed that HuR translocation from the nucleus to the cytoplasm was inhibited, leading to decreased COX-2 and Bax expression levels. Conclusions: TUG1 knockout reduces ischemic damage and neuronal apoptosis by inhibiting HuR nucleocytoplasmic shuttling, making TUG1 a potential therapeutic target for ischemic stroke.

Orexin A protects against cerebral ischemia-reperfusion injury by enhancing reperfusion in ischemic cortex via HIF-1α-ET-1/eNOS pathway

The purpose of this study was to investigate the protective effect and underlying mechanism of orexin A on cerebral ischemia-reperfusion injury, specifically through vasodilation mediated by the hypoxia inducible factor-1α (HIF-1α)-Endothelin-1(ET-1)/endothelial nitric oxide synthase (eNOS) pathway. A model of middle cerebral artery occlusion was established in both wild-type SD rats with exogenous orexin A intervention and in orexin A transgenic rats. Neurological deficit scores and cerebral infarction areas were assessed, and ischemic cortical blood flow was monitored. Gene and protein expression levels of HIF-1α, HIF-2α, ET-1, and three types of NOS were detected using real-time RT-qPCR and Western blot analysis, respectively. Additionally, nitric oxide (NO) levels in the cortex were analyzed through biochemical detection methods. Orexin A demonstrated a protective effect by reducing cerebral infarction and improving neurological deficits, which was achieved by increasing cortical blood flow during reperfusion. This protective mechanism was associated with upregulated HIF-1α expression, downregulated ET-1 expression, upregulated eNOS expression, and increased NO production. This study demonstrates the protective effect of orexin A on cerebral ischemia-reperfusion injury, achieved by regulating the release of vasomotor substances to enhance cortical blood flow during reperfusion. These findings suggest that orexin A may represent a potential therapeutic strategy for ischemic stroke.

Study on the Neuroprotective Mechanism of Salidroside Modified with Polydopamine Nanoparticles in Regulating Notch Pathway by Inhibiting Mir-155 in Stroke Rats

Background and Purpose Stroke threatens neurological function. Salidroside is widely used to treat neurological diseases such as stroke. Previous studies have shown that miR-155 plays an important regulatory role in the pathogenesis of stroke and that the Notch pathway plays an important role in nervous system development and regeneration. Methods PDA NPs-SAL complex was prepared, a stroke rat model was constructed, and the cerebral infarction area of rats was measured. Analyze the changes of PDA NPs-SAL on rat brain tissue and evaluate the neuroprotective effect. Detect the changes in miR-155 and Notch pathway-related proteins in stroke rat models to further understand the role of miR-155 and Notch pathway in the pathogenesis of stroke and their relationship. Results Salidroside had certain protective effects on the nerves of stroke rats, and the effect of PDA NPs-SAL was more prominent. At the same time, PDA NPs-SAL promotes the Notch pathway by inhibiting miR-155, thereby exerting a neuroprotective effect on stroke rats. Conclusion Under the intervention of PDA NPs-SAL, the neurological function scores of rats were reduced, and the effect of PDA NPs-SAL was more significant. In addition, PDA NPs-SAL inhibited miR-155 and activated the Notch pathway, thereby reducing the water content of rat brain tissue and cerebral infarction area, thereby exerting a neuroprotective effect on stroke rats.

Real-time detection of cerebral edema in mice based on low-field nuclear magnetic resonance

Cerebral edema is a secondary symptom of several conditions, including stroke, liver, failure, and severe craniocerebral injury. This study aimed to propose a Low-Field Nuclear Magnetic Resonance (LF-NMR) method for rapid and nondestructive measurement of brain water content and water phase state evaluating animal brain edema models. The water transverse relaxation time (T2) distribution in brain tissues was ascertained by measuring the T2 spectrograms of mouse brain tissues following various drying durations, in accordance with the shifting law of T2 peaks. A methodological study was carried out for the determination of water content in mouse brain tissue by the LF-NMR Peak Area (LF-NMR-PA) method using CuSO4 solution as standard solution. This method was studied in comparison with the wet/dry weight method and the Partial Least Squares Regression (PLSR) prediction method in normal and MCAO model mice, and further applied to the study of mannitol treatment of cerebral edema to validate the precision and accuracy of the hydration measurements as well as the method’s impact on subsequent experiments of cerebral infarct area measurement. In the T2 spectra of mouse brain tissues, the separate peaks T21 (0–10 ms), T22 (10–100 ms), and T23 (100–1000 ms) correspond to fat and bound water, water, and free water in brain tissue, respectively. The LF-NMR-PA method had good specificity, linearity, precision, stability, reproducibility, and recovery. The accuracy and range of use of the LF-NMR-PA method were better than those of the PLSR method, and its results were not significantly different from those of the wet/dry weight method. However, the short measurement time of the LF-NMR-PA method ensured the integrity of the tissues, and the determination of the moisture by this method did not have a significant effect on the subsequent determination of the area of cerebral infarction, which ensured the consistency of the experimental results.

Buqi-Huoxue-Tongnao decoction drives gut microbiota-derived indole lactic acid to attenuate ischemic stroke via the gut-brain axis

BackgroundIschemic stroke belongs to “apoplexy” and its pathogenesis is characterized by qi deficiency and blood stasis combining with phlegm-damp clouding orifices. Buqi-Huoxue-Tongnao decoction (BHTD) is a traditional Chinese medicine formula for qi deficiency, blood stasis and phlegm obstruction syndrome. However, its efficacy and potential mechanism on ischemic stroke are still unclear. This study aims to investigate the protective effect and potential mechanism of BHTD against ischemic stroke.Materials and methodsMiddle cerebral artery occlusion (MCAO) surgery was carried out to establish an ischemic stroke model in rats. Subsequently, the rats were gavaged with different doses of BHTD (2.59, 5.175, 10.35 g/kg) for 14 days. The protective effects of BHTD on the brain and gut were evaluated by neurological function scores, cerebral infarction area, levels of brain injury markers (S-100B, NGB), indicators of gut permeability (FD-4) and bacterial translocation (DAO, LPS, D-lactate), and tight junction proteins (Occludin, Claudin-1, ZO-1) in brain and colon. 16S rRNA gene sequencing and metabolomic analysis were utilized to analyze the effects on gut microecology and screen for marker metabolites to explore potential mechanisms of BHTD protection against ischemic stroke.ResultsBHTD could effectively mitigate brain impairment, including reducing neurological damage, decreasing cerebral infarction and repairing the blood–brain barrier, and BHTD showed the best effect at the dose of 10.35 g/kg. Moreover, BHTD reversed gut injury induced by ischemic stroke, as evidenced by decreased intestinal permeability, reduced intestinal bacterial translocation, and enhanced intestinal barrier integrity. In addition, BHTD rescued gut microbiota dysbiosis by increasing the abundance of beneficial bacteria, including Turicibacter and Faecalibaculum. Transplantation of the gut microbiota remodeled by BHTD into ischemic stroke rats recapitulated the protective effects of BHTD. Especially, BHTD upregulated tryptophan metabolism, which promoted gut microbiota to produce more indole lactic acid (ILA). Notably, supplementation with ILA by gavage could alleviate stroke injury, which suggested that driving the production of ILA in the gut might be a novel treatment for ischemic stroke.ConclusionBHTD could increase gut microbiota-derived indole lactic acid to attenuate ischemic stroke via the gut-brain axis. Our current finding provides evidence that traditional Chinese medicine can ameliorate central diseases through regulating the gut microbiology.

The expression profile of brain-derived exosomal miRNAs reveals the key molecules responsible for spontaneous motor function recovery in a rat model with permanent middle cerebral artery occlusion.

The analysis of alterations in the expression and functionality of brain-derived exosomal miRNAs within ischemic stroke lesions provides significant insights into the mechanisms that contribute to disease recovery. We assessed spontaneous motor function in a rat model of permanent middle cerebral artery occlusion (pMCAO) using motor function scores and magnetic resonance imaging (MRI). Brain-derived exosomes from the infarcted brain tissue of the animal model were extracted and high-throughput sequencing of them was performed followed by bioinformatics analysis for differentially expressed miRNAs target genes. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to measure expression levels of differentially expressed miRNAs at various time points. The oxygen-glucose deprivation (OGD) model was established to investigate gene function through the assessment of cell proliferation and apoptosis using EdU proliferation and JC-1 apoptosis assay. The rat model demonstrated a spontaneous recovery of motor function and a reduction in cerebral infarction area from day 1 to day 14 post-operation. Over the course of the recovery period, miR-24-3p, miR-129-1-3p, and miR-212-5p maintained consistent expression levels, reaching their peak on the initial day following surgery. In the cell model, EdU detection indicated that miR-129-1-3p promoted cellular proliferation, while JC-1 detection revealed its suppressive impact on cellular apoptosis. The current research findings indicated the presence of spontaneous motor function restoration in a rat model of ischemic stroke. MiR-24-3p, miR-129-1-3p, and miR-212-5p were identified as pivotal genes in this recovery process, with miR-129-1-3p potentially influencing the restoration of spontaneous motor function in ischemic stroke through the regulation of neuronal proliferation and apoptosis.

Plasma Levels of Neuron/Glia-Derived Apoptotic Bodies, an In Vivo Biomarker of Apoptosis, Predicts Infarct Growth and Functional Outcome in Patients with Ischemic Stroke.

Evidence demonstrating the involvement of apoptosis in the death of the potentially salvageable area (penumbra zone) in patients during stroke remains limited. Our aim was to investigate whether apoptotic processes occur in penumbral brain tissue by analyzing circulating neuron- and glia-derived apoptotic bodies (CNS-ApBs), which are vesicles released into the bloodstream during the late stage of apoptosis. We have also assessed the clinical utility of plasma neuronal and glial apoptotic bodies in predicting early neurological evolution and functional outcome. The study included a total of 71 patients with acute hemispheric ischemic stroke (73 ± 10years; 30 women). Blood samples were collected from these patients immediately upon arrival at the hospital (within 9h) and at 24 and 72h after symptom onset. Subsequently, isolation, quantification, and phenotypic characterization of CNS-ApBs during the first 72h post-stroke were performed using centrifugation and flow cytometry techniques. We found a correlation between infarct growth and final infarct size with the amount of plasma CNS-ApBs detected in the first 72h after stroke. In addition, patients with neurological worsening (progressive ischemic stroke) had higher plasma levels of CNS-ApBs at 24h after symptom onset than those with a stable or improving course. Circulating CNS-ApB concentration was further associated with patients' functional prognosis. In conclusion, apoptosis may play an important role in the growth of the cerebral infarct area and plasma CNS-ApB quantification could be used as a predictive marker of penumbra death, neurological deterioration, and functional outcome in patients with ischemic stroke.

Huanglian Jiedu decoction alleviates ischemia-induced cerebral injury in rats by mitigating NET formation and activiting GABAergic synapses.

Huanglian Jiedu decoction (HLJD) has been used to treat ischemic stroke in clinic. However, the detailed protective mechanisms of HLJD on ischemic stroke have yet to be elucidated. The aim of this study is to elucidate the underlying pharmacological mechanisms of HLJD based on the inhibition of neuroinflammation and the amelioration of nerve cell damage. A middle cerebral artery occlusion reperfusion (MCAO/R) model was established in rats and received HLJD treatment. Effects of HLJD on neurological function was assessed based on Bederson's score, postural reflex test and asymmetry score. 2, 3, 5-Triphenyltetrazolium chloride (TTC) staining, Hematein and eosin (HE) and Nissl staining were used to observe the pathological changes in brain. Then, transcriptomics was used to screen the differential genes in brain tissue in MCAO/R model rats following HLJD intervention. Subsequently, the effects of HLJD on neutrophil extracellular trap (NET) formation-related neuroinflammation, gamma-aminobutyric acid (GABA)ergic synapse activation, nerve cell damage and proliferation were validated using immunofluorescence, western blot and enzyme-linked immunosorbent assay (ELISA). Our results showed that HLJD intervention reduced the Bederson's score, postural reflex test score and asymmetry score in MCAO/R model rats. Pathological staining indicated that HLJD treatment decreased the cerebral infarction area, mitigated neuronal damage and increased the numbers of Nissl bodies. Transcriptomics suggested that HLJD affected 435 genes in MCAO/R rats. Among them, several genes involving in NET formation and GABAergic synapses pathways were dysregulated. Subsequent experimental validation showed that HLJD reduced the MPO+CitH3+ positive expression area, reduced the protein expression of PAD4, p-P38/P38, p-ERK/ERK and decreased the levels of IL-1β, IL-6 and TNF-α, reversed the increase of Iba1+TLR4+, Iba1+p65+ and Iba1+NLRP3+ positive expression area in brain. Moreover, HLJD increased GABA levels, elevated the protein expression of GABRG1 and GAT3, decreased the TUNEL positive expression area and increased the Ki67 positive expression area in brain. HLJD intervention exerts a multifaceted positive impact on ischemia-induced cerebral injury in MCAO/R rats. This intervention effectively inhibits neuroinflammation by mitigating NET formation, and concurrently improves nerve cell damage and fosters nerve cell proliferation through activating GABAergic synapses.

Impact of acupuncture on ischemia/reperfusion injury: Unraveling the role of miR-34c-5p and autophagy activation

We have previously reported that the expression of miR-34c-5p was up-regulated during acupuncture treatment in the setting of a cerebral ischemia/reperfusion injury (CIRI), indicating that miR-34c-5p plays an important role in healing from a CIRI-induced brain injury. This study sought to evaluate the effects of acupuncture on miR-34c-5p expression and autophagy in the forward and reverse directions using a rat focal cerebral ischemia/reperfusion model. After 120 minutes of middle cerebral artery occlusion and reperfusion, rats were treated with acupuncture at the "Dazhui" (DU20), "Baihui" (DU26) and "Renzhong" (DU14) points. Neurologic function deficit score, cerebral infarct area ratio, neuronal apoptosis and miR-34c-5p expression were evaluated 72 hr after treatment. The autophagy agonist RAPA and the antagonist 3MA were used to evaluate the neuro protective effects of autophagy-mediated acupuncture. We found that acupuncture treatment improved autophagy in the brain tissue of CIRI rats. Acupuncture reversed the negative effects of 3MA on CIRI, and acupuncture combined with RAPA further enhanced autophagy. We also found that acupuncture could increase miR-34c-5p expression in hippocampal neurons after ischemia/reperfusion. Acupuncture and a miR-34c agomir were able to enhance autophagy, improve neurologic deficits, and reduce the cerebral infarct area ratio and apoptosis rate by promoting the expression of miR-34c-5p. Silencing miR-34c resulted in a significantly reduced activating effect of acupuncture on autophagy and increased apoptosis, neurologic deficit symptoms, and cerebral infarct area ratio. This confirms that acupuncture can upregulate miR-34c-5p expression, which is beneficial in the treatment of CIRI.

Prevalence and Radiographic Characteristics of Cerebral Infarction after Surgery in Patients with Glioma: A Retrospective Study.

The aim of our study was to analyze risk factors for postoperative cerebral infarction in patients with glioma in our hospital, and to compare medical imaging techniques for early diagnosis of postoperative cerebral infarction. A retrospective analysis was conducted on 178 patients (male: 78, female: 100) who underwent glioma surgery at our hospital between May 2015 and October 2023. They were divided into two groups based on the presence of postoperative cerebral infarction within 7 days: the cerebral infarction group (n = 85) and the non-cerebral infarction group (n = 93). Magnetic resonance imaging (MRI) was used to assess the location, distribution, and volume of the tumor before surgery. During the perioperative period, patient postoperative time, intraoperative blood loss, and other relevant data were documented. Computed tomography perfusion (CTP) and diffusion-weighted imaging (DWI) imaging techniques were employed to evaluate the occurrence, area, location, and shape of cerebral infarction. The imaging characteristics of postoperative cerebral infarction were noted. Apparent diffusion coefficient values, apparent diffusion coefficient (ADC) of whole-brain CTP parameters, cerebral blood flow (CBF), cerebral blood volume (CBV), time to peak (TTP), mean transit time (MTT), and DWI parameters were measured. The sensitivity and specificity of CTP, DWI, and their combined diagnosis for postoperative cerebral infarction were compared, with consistency assessed using the Kappa value. This study found that 85 patients (47.8%) experienced postoperative cerebral infarction. Significant risk factors included tumor location in the temporal lobe, tumor volume ≥23.57 cm3, number of surgeries >1, World Health Organization (WHO) grade >3, and intraoperative blood loss >79.83 mL (p < 0.05). Imaging examinations revealed that CTP combined with DWI diagnosis detected cerebral infarctions in 84 patients, showing lower CBF and CBV, and higher TTP, and MTT in the infarct group (p < 0.05). The Kappa values for CTP, DWI, and the combined diagnosis were 0.762, 0.833, and 0.937, respectively (p < 0.001). The prevalence of cerebral infarction in patients with glioma is high and is affected by many factors. Timely imaging examination can detect and predict the occurrence of cerebral infarction in patients after surgery, which is of great significance for improving the prognosis of patients.