The protective effects of Axitinib on blood-brain barrier dysfunction and ischemia-reperfusion injury in acute ischemic stroke.

BACKGROUND AND PURPOSE: The pathophysiological features of acute ischemic stroke (AIS) often involve dysfunction of the blood-brain barrier (BBB), characterized by the degradation of tight junction proteins (Tjs) leading to increased permeability. This dysfunction can exacerbate cerebral injury and contribute to severe complications. The permeability of the BBB fluctuates during different stages of AIS and is influenced by various factors. Developing effective therapies to restore BBB function remains a significant challenge in AIS treatment. High levels of vascular endothelial growth factor (VEGF) in the early stages of AIS have been shown to worsen BBB breakdown and stroke progression. Our study aimed to investigate the protective effects of the VEGF receptor inhibitor Axitinib on BBB dysfunction and cerebral ischemia/reperfusion-induced injury. METHODS: BEnd3 cell exposed to oxygen-glucose deprivation (OGD) model was constructed to estimate pharmacological activity of Axitinib (400 ng/ml) on anti-apoptosis and pathological barrier function recovery. In vivo, rats were subjected to a 1 h transient middle cerebral artery occlusion and 23 h reperfusion (tMCAO/R) to investigate the permeability of BBB and cerebral tissue damage. Axitinib was administered through the tail vein at the beginning of reperfusion. BBB integrity was assessed by Evans blue leakage and the expression levels of Tjs claudin-5 and occludin. RESULTS: Our research revealed that co-incubation with Axitinib enhanced the cell viability of OGD-insulted bEnd3 cells, decreased LDH leakage rate, and suppressed the expression of apoptosis-related proteins cytochrome C and Bax. Axitinib also mitigated the damage to Tjs and facilitated the restoration of transepithelial electrical resistance in OGD-insulted bEnd.3 cells. In vivo, Axitinib administration reduced intracerebral Evans blue leakage and up-regulated the expression of Tjs in the penumbra brain tissue in tMCAO/R rats. Notably, 10 mg/kg Axitinib exerted a significant anti-ischemic effect by decreasing cerebral infarct volume and brain edema volume, improving neurological function, and reducing pro-inflammatory cytokines IL-6 and TNF-α in the brain. CONCLUSIONS: Our study highlights Axitinib as a potent protectant of blood-brain barrier function, capable of promoting pathological blood-brain barrier recovery through VEGF inhibition and increased expression of tight junction proteins in AIS. This suggests that VEGF antagonism within the first 24 h post-stroke could be a novel therapeutic approach to enhance blood-brain barrier function and mitigate ischemia-reperfusion injury.

Acute cerebral ischemic stroke as a presentation of polycythemia vera: a case report and a review of literature.

Polycythemia vera (PV) is a disease of stem cells characterized by pan hyperplastic, malignant, and neoplastic bone marrow conditions. It is characterized by an increased absolute red blood cell count due to uncontrolled red blood cell synthesis, as well as excessive white blood cell and platelet production. Although the relationship between PV and stroke, especially ischemic stroke, is widely known around the world, no previous cases have been reported from Somalia. Case presentation: In the presenting study, we report a 60-year-old male patient who presented with a right-side weakness for 3 days. After laboratory and brain imaging, he was diagnosed with an acute cerebral infarct affecting the left basal ganglion secondary to PV. Conclusions: PV as the cause of ischemic stroke is a rare condition but can be encountered in clinical practice, and clinicians should be familiar with this combination.

Increased NOX2 expression in astrocytes leads to eNOS uncoupling through dihydrofolate reductase in endothelial cells after subarachnoid hemorrhage.

Introduction: Endothelial nitric oxide synthase (eNOS) uncoupling plays a significant role in acute vasoconstriction during early brain injury (EBI) after subarachnoid hemorrhage (SAH). Astrocytes in the neurovascular unit extend their foot processes around endothelia. In our study, we tested the hypothesis that increased nicotinamide adenine dinucleotide phosphate oxidase 2 (NOX2) expression in astrocytes after SAH leads to eNOS uncoupling. Methods: We utilized laser speckle contrast imaging for monitoring cortical blood flow changes in mice, nitric oxide (NO) kits to measure the level of NO, and a co-culture system to study the effect of astrocytes on endothelial cells. Moreover, the protein levels were assessed by Western blot and immunofluorescence staining. We used CCK-8 to measure the viability of astrocytes and endothelial cells, and we used the H2O2 kit to measure the H2O2 released from astrocytes. We used GSK2795039 as an inhibitor of NOX2, whereas lentivirus and adeno-associated virus were used for dihydrofolate reductase (DHFR) knockdown in vivo and in vitro. Results: The expression of NOX2 and the release of H2O2 in astrocytes are increased, which was accompanied by a decrease in endothelial DHFR 12 h after SAH. Moreover, the eNOS monomer/dimer ratio increased, leading to a decrease in NO and acute cerebral ischemia. All of the above were significantly alleviated after the administration of GSK2795039. However, after knocking down DHFR both in vivo and in vitro, the protective effect of GSK2795039 was greatly reversed. Discussion: The increased level of NOX2 in astrocytes contributes to decreased DHFR in endothelial cells, thus aggravating eNOS uncoupling, which is an essential mechanism underlying acute vasoconstriction after SAH.

Neurointerventional infusion of hemoglobin oxygen carrier prevents brain damage from acute cerebral ischemia in rats.

Aim: To save brain cells in acute cerebral infarction by injecting hemoglobin oxygen carrier (HBOC) into the blood vessel blockage of the cerebral infarction site through a microcatheter. Methods: 120 male rats were divided into four groups: control (CTRL), ischemia (I), ischemia + low perfusion (I + LP), and ischemia + high perfusion (I + HP). Perfusion groups (ischemia, I + LP, and I + HP) underwent MCAO surgery with intraluminal monofilament. These groups were subdivided into 6 h, 12 h, and 24 h (n = 10/group). RT-PCR, Western-Blot, immunohistochemistry, and apoptosis assays were used to detect apoptosis, hypoxia range and extent, and ischemia. Results: Compared with the I group, the neurological deficit sign scores of the I + HP group were statistically significant at 12 h. Compared with the I group, the neurological deficit sign scores of the I + LP group and the I + HP group were statistically significant at 24 h. At all time points, compared with the I group and the I + LP group, Caspase-3, HIF-1α, and Cytochrome C protein levels were significantly decreased in the I + HP group. Bcl-2 and BAX mRNA levels were also significantly decreased in the same group. TNF-α, IL-6, and IL-1ß cytokines were significantly decreased in the I + HP group as well. The infarct size of rats in the I + HP group was smaller than that of the I + LP group, which was smaller than ischemia alone. Time of perfusion had an obvious effect as infarct size was smaller with longer perfusion. The number of Nissl stained cells in the I + HP group was increased compared with the ischemia and the I + LP group, and was proportional to the time of perfusion. Conclusion: Time- and rate-controlled perfusion of HBOC to acutely occluded cerebral vascular regions through microcatheters can effectively protect ischemic brain tissue in rats.

PRIMED2 Preclinical Evidence Scoring Tool to Assess Readiness for Translation of Neuroprotection Therapies.

Many neuroprotective and other therapies for treatment of acute ischemic stroke have failed in translation to human studies, indicating a need for more rigorous, multidimensional quality assessment of the totality of preclinical evidence supporting a therapy prior to conducting human trials. A consensus panel of stroke preclinical model and human clinical trial experts assessed candidate items for the translational readiness scale, compiled from prior instruments (STAIR, ARRIVE, CAMARADES, RoB 2) based on importance, reliability, and feasibility. Once constructed, the tool was applied by two independent raters to four current candidate acute stroke therapies, including two pharmacologic agents [nerinetide and trans-sodium crocetinate] and two device interventions [cathodal transcranial direct current stimulation and fastigial nucleus stimulation]. The Preclinical evidence of Readiness In stroke Models Evaluating Drugs and Devices (PRIMED2) assessment tool rates the totality of evidence available from all reported preclinical animal stroke model studies in 11 domains related to diversity of tested animals, time windows, feasibility of agent route of delivery, and robustness of effect magnitude. Within each content domain, clearly operationalized rules assign strength of evidence ratings of 0-2. When applied to the four assessed candidate agents, inter-rater reliability was high (kappa = 0.88), and each agent showed a unique profile of evidentiary strengths and weaknesses. The PRIMED2 assessment tool provides a multidimensional assessment of the cumulative preclinical evidence for a candidate acute stroke therapy on factors judged important for successful basic-to-clinical translation. Further evaluation and refinement of this tool is desirable to improve successful translation of therapies for acute stroke.

Omentin Is Independently Associated with Stroke Severity and Ipsilateral Carotid Artery Stenosis in Patients with Acute Cerebral Ischemia.

Mounting evidence indicates an association between adipokines and inflammation-related atherosclerosis. Here, we sought to investigate the association of vaspin and omentin with clinical characteristics and outcomes of patients with acute cerebral ischemia (ACI). Consecutive ACI patients were evaluated within 24 h from symptom-onset. Stroke aetiology was classified using TOAST criteria. Adipokines were assayed using quantikine enzyme immunoassay commercially available kits. Stroke severity was assessed by NIHSS-score, and ipsilateral carotid stenosis (≥50% by NASCET criteria) by ultrasound and CT/MR angiography. Major cerebrovascular events were assessed at three months. We included 135 ACI patients (05 (78%) and 30 (22%) with acute ischemic stroke and transient ischemic attack, respectively; mean age ± SD: 59 ± 10 years; 68% men; median NIHSS-score: 3 (IQR:1-7)). Omentin was strongly correlated to admission stroke severity (Spearman rho coefficient: +0.303; p < 0.001). Patients with ipsilateral carotid stenosis had higher omentin levels compared to patients without stenosis (13.3 ± 8.9 ng/mL vs. 9.5 ± 5.5 ng/mL, p = 0.014). Increasing omentin levels were independently associated with higher stroke severity (linear regression coefficient = 0.290; 95%CI: 0.063-0.516; p = 0.002) and ipsilateral carotid stenosis (linear regression coefficient = 3.411; 95%CI: 0.194-6.628; p = 0.038). No association of vaspin with clinical characteristics and outcomes was found. Circulating omentin may represent a biomarker for the presence of atherosclerotic plaque, associated with higher stroke severity in ACI patients.

Analysis of regulatory effect of miR-149-5p on Sphingosine-1-phosphate receptor 2 of pericytes and its neuroprotective molecular mechanism after acute cerebral ischemia reperfusion in rats.

To investigate the effect of miR-149-5p on sphingosine-1-phosphate receptor 2 (S1PR2) expression level and contents of matrix metalloproteinase (MMP-9) and superoxide dismutase (SOD) in the pericytes after acute cerebral ischemia reperfusion in rats, so as to clarify the neuroprotective molecular mechanism induced by miR-149-5p and provide references for the treatment of neurological diseases, 60 male SD rats aged 7-8 weeks were selected and divided randomly into test group (establishing middle cerebral artery occlusion (MCAO) model) and control group (no modeling). Rat pericytes and peripheral cerebral infarction tissues were collected 12 h, 1 d, 3 d, 5 d, and 7 d after MCAO modeling, respectively. The pericytes were identified by immunofluorescence assay (IFA) and transfected with miR-149-5p. Fluorescence quantitative PCR (FQPCR) and Western blot were adopted to detect S1PR2 expression level. The expression of S1PR2 in MCAO model rats was detected by IFA. Immunohistochemistry (IHC) and quantitative real-time PCR (qRT-PCR) were used to detect the changes of MMP9 protein and mRNA levels of SOD1, SOD2, and SOD3 in brain tissue. The results showed that mRNA level and protein expression level of S1PR2 in the test group were higher than those in the control group three days after MCAO modeling (P < 0.05); the expression of S1PR2 increased 12 h after MCAO modeling and returned to the normal level on the 5th day, and the content of MMP9 protein in brain tissue of the test group was significantly lower than that of the control group (P < 0.05); the mRNA levels and SODs activity of SOD1, SOD2, and SOD3 in the test group were higher than those in the control group (P < 0.05). Therefore, miR-149-5p played a neuroprotective role by regulating S1PR2 to change the expression levels of SODS and MMP9.

Effect of vitexin on Nrf2/ARE pathway in alleviating oxidative stress in rats with acute cerebral ischemia-reperfusion / 中草药

Objective: To investigate the effect of vitexin on oxidative stress in rats with acute cerebral ischemia-reperfusion by regulating the pathway of nuclear factor E2-related factor 2 (Nrf2)/antioxidant response element (ARE). Methods: A total of 54 SD rats were randomly divided into Sham group, model group, positive control (edaravone 0.56 mg/kg) group and vitexin low, medium, and high dose (10, 20, 40 mg/kg) groups. The rat models with acute cerebral ischemia were established except the Sham group. After reperfusion, rats in Sham group and model group were received ip saline. The edaravone group and vitexin groups were administered according to the corresponding dose, once every 8 h for a total of three times. The neurobehavioral scores of rats before and after intervention were compared. HE staining was used to detect the pathological changes of cerebral cortex. The levels of MDA, NO, SOD, and GSH in cerebral cortex of rats were detected by the kit. The mRNA and protein expression levels of Nrf2/ARE pathway related gene in rat cerebral cortex were detected by real-time fluorescent quantitative PCR (qRT-PCR) and Western blotting. Results: Before and after intervention, there was no significant change in the neurobehavioral score of rats in the Sham group, the model group was higher than before intervention (P < 0.01), the edaravone group and the vitexin groups were lower than before intervention (P < 0.01), and there was a significant difference in the neurobehavioral score between the groups after intervention (P < 0.01). In the Sham group, the distribution of nerve cells was uniform and dense, and the morphology of cell body and nucleus was normal. In the model group, edaravone group and vitexin group, the arrangement of brain tissue was disordered and loose, in which liquefying necrosis and disappearance of cell structure were observed in the model group; in the low-dose group, the arrangement of cells was seriously disordered and loose. In the vitexin medium dose group and edaravone group, the area of liquefying necrosis was small, and most of the cells were normal; In the vitexin high dose group, only a few liquefying necrosis were found, the cell structure was basically normal, and the cell arrangement was slightly disordered. Compared with the Sham group, the levels of MDA and NO in the cerebral cortex of the model group were significantly increased (P < 0.01), the levels of SOD and GSH were significantly reduced (P < 0.01), and the expression levels of Nrf2 and γ-GCS mRNA were significantly increased (P < 0.01), the expression of cytoplasmic Nrf2 and γ-GCS proteins were significantly increased (P < 0.01), and the expression levels of nuclear Nrf2 and HO-1 proteins were significantly decreased (P < 0.01). Compared with the model group, the levels of MDA and NO in the cerebral cortex of rats in the edaravone group and low, medium and high dose groups of vitexin were significantly reduced (P < 0.01), and the levels of SOD and GSH were significantly increased (P < 0.01), Nrf2 and γ-GCS mRNA expression levels were significantly reduced (P < 0.01), cytoplasmic Nrf2, γ-GCS protein expressions were significantly reduced (P < 0.01), and nuclear Nrf2 and HO-1 protein expression levels were significantly increased (P < 0.01). And the levels of MDA, NO, SOD, GSH and Nrf2, γ-GCS, HO-1 mRNA and protein expression in rat cerebral cortex in each dose group were dose-dependent, with significant differences between groups (P < 0.01). Conclusion: Vitexin can alleviate oxidative stress in rats with acute cerebral ischemia-reperfusion. It is speculated that it is related to the regulation of Nrf2/ARE signaling pathway, the up-regulation of Nrf2 gene and protein expression, the promotion of its movement from cytoplasm to nucleus, the up-regulation of HO-1 expression, the inhibition of γ-GCS expression, and the enhancement of the body’s ability to response to oxidative stress.

[Scalp-acupuncture improves neurological function by regulating expression of IL-10 mRNA, IL-6 mRNA and TNF-α of parahippocampal gyrus in cerebral ischemic rats].

OBJECTIVE: To observe the effect of scalp-acupuncture intervention on the expression of Interleukin (IL)-10 mRNA, IL-6 mRNA and tumor necrosis factor (TNF) - α in the parahippocampal gyrus of cerebral ischemia (CI) rats, so as to explore its molecular mechanisms underlying improvement of CI. METHODS: A total of 64 male SD rats were randomized into normal control, model, medication and scalp-acupuncture groups (n＝16 rats in each group). The focal CI model was established by middle cerebral artery occlusion. Intraperitoneal injection of Ammonium Pyrrolidine Dithiocarbamate (100 mg•kg－1•d－1) was administrated for rats in the medication group, once a day for 7 days. For rats of the scalp-acupuncture group, the acupuncture needles were rapidly inserted into bilateral Dingnieqianxiexian (MS6), followed by twirling the needles at 100 cycles/min for 1 min, once again every 10 min during 20 min' needle retention. The treatment was conducted once a day for 7 days. The neurologic deficit score (0－4 points, impaired consciousness, death, etc.) and neurological function score (motor, sensory and sensory tests, 0－10 points) were assessed according to Longa's (1989) and Schabitz's (2004) methods, respectively. The expression levels of IL-10 mRNA and IL-6 mRNA were detected by fluorescence quantitative PCR, and the expression of TNF-α was detected by immunohistochemistry. RESULTS: After modeling, the neurologic deficit and neurological function scores and the expression levels of IL-10 mRNA, IL-6 mRNA and TNF-α protein in the parahippocampus were significantly increased in the model group than in the normal control group (P<0.01). Following the intervention, the neurologic deficit and neurological function scores as well as IL-6 mRNA and TNF-α protein expression were significantly down-regulated in both scalp-acupuncture and medication groups (P<0.05), and the expression of IL-10 mRNA was obviously increased (P<0.05) relevant to the model group. CONCLUSION: Scalp-acupuncture can improve neurologic function in CI rats, which is related to its effects in up-regulating the expression of IL-10, then down-regulating the expression of IL-6 and TNF-α (reducing inflammatory response) in the parahippocampal gyrus.

Autonomic Cardiac Regulation in Experimental Comorbidity of Chronic Obstructive Pulmonary Disease and Acute Cerebral Ischemia.

Autonomic regulation of the heart was examined in 5 groups of rats: intact, sham-operated, experimental chronic obstructive pulmonary disease, acute cerebral ischemia, and acute cerebral ischemia modeled against the background of chronic obstructive pulmonary disease. The latter was provoked by combination of inhaled papain and intraperitoneal bacterial LPS, whereas acute cerebral ischemia was modeled by single-stage bilateral occlusion of the common carotid arteries. Chronic obstructive pulmonary disease was verified by X-ray computed microtomography. The disturbances in autonomic control of the heart during comorbid pathologies were most prominent; they were manifested by overstrain and decompensation of the mechanisms implicated in the heart control and systolic-diastolic arterial hypotension. The correlations were established between blood oxygenation, respiration rate, and some parameters of autonomic cardiac regulation. The data attest to relevance and usefulness of the developed model of respiratory and cerebrovascular comorbidity in assessment of pathophysiological mechanisms underlying dysregulation of the heart and the development of personalized approaches for its pharmacological correction.

Timing of Decompressive Craniectomy for Malignant Middle Cerebral Artery Infarction: A Single-Center Analysis.

Background and aim: Malignant middle cerebral artery infarction (MMCAI) usually leads to brain edema that may result in transtentorial herniation and brainstem compression. The prognosis of MMCAI is generally poor. The aim of this study was to discuss our experience with surgical decompression for MMCAI, and determine the association between timing of craniectomy and neurological outcomes. Methods: We identified consecutive patients diagnosed with MMCAI who underwent decompressive craniectomy (DC). Clinical and demographic data were obtained from electronic medical records, including: age, sex, preoperative Glasgow Coma Scale (GCS) score, surgery timing, postoperative GCS scores, and modified Rankin Scale (mRS) scores. Results: This study included 27 stroke patients (aged 38⁻80 years) operated within 72 h of the onset of neurological symptoms. Sixteen, five, and six patients underwent DC within 24 h, between 24 and 48 h, and after 48 h after onset of symptoms, respectively. Five patients died after the surgery. Patients who underwent DC within 24 h and 24⁻48 h had better mean GCS scores than those who underwent DC after 48 h (p = 0.000, p = 0.015). In addition, patients who underwent DC within 24 h had better mean postoperative mRS scores (p = 0.000) than other patients. Patients older than 60 years had significantly lower GCS scores (p = 0.027) and higher mRS scores (p = 0.033) than younger patients. Conclusion: Our findings support that DC had satisfying outcomes in patients who underwent DC within 24 h. Older age and lower Glasgow Coma Scale scores among DC patients with MMCAI are associated with high morbidity and mortality.

Wogonin improves functional neuroprotection for acute cerebral ischemia in rats by promoting angiogenesis via TGF-ß1.

BACKGROUND: Previous studies showed that wogonin is a potential candidate for more effective treatment of neuronal and inflammatory disease and could offer neuroprotective activity in various models, but all these studies were in vitro. Our research aimed to investigate the neuroprotective effect of wogonin on focal cerebral ischemia in rats and uncover its potential mechanism. METHODS: A total of 80 male SD rats were randomly divided into a sham operation group (Sham group, 20 rats), a normal saline group (NS group, 20 rats), and a wogonin intervention group (W2W group, 20 rats), while the remaining 20 rats were kept as a substitute. The model of focal cerebral ischemia (MCAO) was established by thread embolization. The neurological deficits were evaluated by the modified neurological deficit scale (mNSS). The laser confocal technique was used to observe the diameter, density, and total area of microvessel. Lastly, the expression of transforming growth factor-ß1 (TGF-ß1) was detected by Western blot. RESULTS: The mNSS scores of the NS group and Wn2W group were 6.57±1.13 and 4.39±0.92 respectively, and the difference between NS group and Wn2W group was statistically significant (P<0.05); the vascular diameter of the Wn2W group, Sham group, and NS group were 2.93±0.19, 4.24±0.16, and 3.56±0.22 µm respectively, and the differences among these groups were statistically significant (F=102.142, P<0.01). Furthermore, the differences in the vascular density (F=290.49, P<0.01) and total microvessel area (F=163.08, P<0.01) among these groups were also statistically significant. The expression of TGF-ß1 in ischemic brain tissue of the Sham group, NS group, and Wn2W group were 0.46±0.14, 0.62±0.18, and 0.94±0.21 respectively, and the differences among these groups were statistically significant (F=102.142, P<0.01). CONCLUSIONS: Wogonin can markedly reduce nerve injury and improve nerve function in rats with cerebral ischemia, which may be related to the TGF-ß1 pathway.

Scalp-acupuncture improves neurological function by regulating expression of IL-10 mRNA, IL-6 mRNA and TNF-α of parahippocampal gyrus in cerebral ischemic rats / 针刺研究

OBJECTIVE: To observe the effect of scalp-acupuncture intervention on the expression of Interleukin (IL)-10 mRNA, IL-6 mRNA and tumor necrosis factor (TNF) - α in the parahippocampal gyrus of cerebral ischemia (CI) rats, so as to explore its molecular mechanisms underlying improvement of CI. METHODS: A total of 64 male SD rats were randomized into normal control, model, medication and scalp-acupuncture groups (n＝16 rats in each group). The focal CI model was established by middle cerebral artery occlusion. Intraperitoneal injection of Ammonium Pyrrolidine Dithiocarbamate (100 mg•kg－1•d－1) was administrated for rats in the medication group, once a day for 7 days. For rats of the scalp-acupuncture group, the acupuncture needles were rapidly inserted into bilateral Dingnieqianxiexian (MS6), followed by twirling the needles at 100 cycles/min for 1 min, once again every 10 min during 20 min' needle retention. The treatment was conducted once a day for 7 days. The neurologic deficit score (0－4 points, impaired consciousness, death, etc.) and neurological function score (motor, sensory and sensory tests, 0－10 points) were assessed according to Longa's (1989) and Schabitz's (2004) methods, respectively. The expression levels of IL-10 mRNA and IL-6 mRNA were detected by fluorescence quantitative PCR, and the expression of TNF-α was detected by immunohistochemistry. RESULTS: After modeling, the neurologic deficit and neurological function scores and the expression levels of IL-10 mRNA, IL-6 mRNA and TNF-α protein in the parahippocampus were significantly increased in the model group than in the normal control group (P<0.01). Following the intervention, the neurologic deficit and neurological function scores as well as IL-6 mRNA and TNF-α protein expression were significantly down-regulated in both scalp-acupuncture and medication groups (P<0.05), and the expression of IL-10 mRNA was obviously increased (P<0.05) relevant to the model group. CONCLUSION: Scalp-acupuncture can improve neurologic function in CI rats, which is related to its effects in up-regulating the expression of IL-10, then down-regulating the expression of IL-6 and TNF-α (reducing inflammatory response) in the parahippocampal gyrus.

Neuroprotective effect of vitexin on acute cerebral ischemia reperfusion rats and its effect on Th1/Th2 drift / 中草药

Objective: To investigate the neuroprotective effect of vitexin (VT) on acute cerebral ischemia (ACI) reperfusion rats and its effect on helper T cell 1 (Th1)/Th2 drift. Methods: Rat models of ACI reperfusion were established and divided into model group, low, medium and high doses of VT (0.94, 1.88, 3.76 mg/kg) groups and sham operated group. After 1 h of ischemia-reperfusion, rats in different doses of VT were given different concentrations of VT by ip, and rats in sham operated group and model group were given the same amount of saline by ip for three consecutive days. The general state of rats was observed. Longa neurological score before and after administration, and morphological changes of neurons in brain tissue after administration were evaluated and compared. The damage rates of single and double strands breaks of DNA in brain tissue were compared. The levels of Th1 and Th2 markers interferon-γ (INF-γ) and interleukin-4 (IL-4) in brain tissue were measured and the INF-γ/IL-4 was calculated. Results: The success rate of ACI reperfusion model was 88.89%. The mental state of rats in the model group was not good, which was improved in the three doses of VT groups after administration. Compared with model group, the neurological function scores of rats in each dose group of VT were significantly decreased 1 d and 3 d after administration (P < 0.05) in a dose-dependent manner. The results of HE showed that the volume of neurons and nucleus of neurons in model group were reduced, and the injury of neurons in VT groups was alleviated after administration, especially in high dose group. Compared with the sham operated group, the damage rates of single and double strand breaks of DNA, INF-γ level and INF-γ/IL-4 in the model group were significantly higher (P < 0.05), while IL-4 level was significantly lower (P < 0.05). Compared with the model group, the rate of DNA single- and double-strand breakage damage, INF-γ level and INF-γ/IL-4 were significantly decreased (P < 0.05) and the level of IL-4 was significantly increased (P < 0.05) in each dose group of VT in a dose-dependent manner. Conclusion: VT has protective effect on nerve function in ACI reperfusion rats, and with the best protective effect at dose of 3.76 mg/kg, which may be related to regulating the balance of Th1/Th2 cells to shift to Th2 and alleviating DNA damage in brain cells.

Effect of Ginkgo biloba extract on TLR4/NF-κB pathway in alleviating cerebral ischemia in mice with acute cerebral ischemia / 中草药

Objective: To observe the effect of Ginkgo biloba extract on cerebral cortical ischemia in mice with acute cerebral ischemia and explore its effect on Toll-like receptor 4 (TLR4)/nuclear factor-κB (NF-κB) pathway. Methods: Sixty adult healthy male CD1 mice were randomly divided into sham operated group, model group, positive control group (3 × 104 IU/kg ulinastatin) and Ginkgo biloba extract low, medium and high doses (10, 20, 40 mg/kg) groups. The acute cerebral ischemia model in other five groups were all established except sham operated group. After successful modeling, mice in each group were given corresponding drugs iv tail, sham operation group and model group were given the same amount of saline. The pathological changes and histological grades of the cerebral cortex were observed. The survival neuron density of each group was compared. Malondialdehyde (MDA) and superoxide dismutase (SOD) were measured by colorimetry and xanthine oxidation respectively. The expressions of Toll like receptor 4 (TLR4) and nuclear transcription factor NF-κB p65 (NF-κB p65) mRNA were detected by real-time fluorescent quantitative PCR (qRT-PCR). Western blotting was used to detect TLR4 and NF-κB p65 protein expression. Results: In sham-operated group, the structure of cerebral cortex was normal, and the cells were full and arranged tightly and orderly. In the blank control group, the structure of cerebral cortex was severely damaged, and the cells were severely shrunk and disordered. Pathological changes in the Ulinastatin group and Ginkgo biloba extract groups were alleviated and dose-dependent. Compared with the sham operation group, the other five groups showed higher histological grade and MDA level in cerebral cortex (P < 0.05). Compared with the model group, the histological grade and MDA level of cerebral cortex decreased in ulinastatin group and Ginkgo biloba extract groups (P < 0.05). Compared with ulinastatin group, the histological grade and MDA content of brain cortex decreased in high dose group (P < 0.05). Compared with the sham operated group, the density of surviving neurons and the level of SOD in the other five groups decreased (P < 0.05). Compared with the model group, the density of survival neurons and the level of SOD in cortex increased in ulinastatin group and Ginkgo biloba extract group (P < 0.05). Compared with ulinastatin group, the density of survival neurons and the level of SOD in cortex increased in high dose group (P < 0.05). Compared with the sham operated group, the expression of TLR4, NF-κB p65 mRNA and protein in the other five groups increased (P < 0.05). Compared with the model group, the expression of TLR4, NF-κB p65 mRNA and protein in cortex of mice in ulinastatin group and Ginkgo biloba extract group decreased (P < 0.05). Compared with ulinastatin group, the expression of TLR4, NF-κB p65 mRNA and protein in the brain cortex of mice decreased in the high dose group (P < 0.05). Conclusion: Ginkgo biloba extract can alleviate the pathological changes caused by cerebral ischemia in mice with acute cerebral ischemia, alleviate neurological impairment, and improve oxidative stress indexes of cerebral cortex tissues. It is presumed that it may be achieved by regulating TLR4/NF-κB p65 signaling pathway and inhibiting the expressions of TLR4 and NF-κB p65 proteins.

Metabonomics Analysis of Sailuotong Capsules in Treatment of Acute Cerebral Ischemia in Rats / 中国实验方剂学杂志

Objective:To explore the mechanism of Sailuotong capsules in treating acute cerebral ischemia from the perspective of metabonomics. Method:A total of 24 SD rats were randomly divided into 3 groups, including sham-operated group, model group and Sailuotong group (33 mg·kg-1). The rat model of acute multiple cerebral infarction was established by injecting fluorescent microspheres into internal carotid artery. After the successful operation, rats in Sailuotong group were administered by duodenal injection immediately, and the dosage volume was 2 mL·kg-1. Endogenous metabolites in rat brain tissues of each group were determined by UPLC-Q-TOF-MS. The relevant data and biomarkers were analyzed by principal component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA). Result:The analysis of pattern recognition indicated that the metabolite profiles in model group and sham-operated group were separated obviously, and ten biomarkers related to acute cerebral ischemia were also identified. Compared with the sham-operated group, contents of N-acetylaspartate (NAA), fumaric acid, glutathione, dehydroascorbic acid, aspartic acid and S-adenosylhomocysteine were decreased, while the contents of arginine, citrulline, saccharopine and hydantoin-5-propionic acid were increased in the model group. Meanwhile, the ten abnormal biomarkers mentioned above got restoration in Sailuotong group. Conclusion:The main regulated metabolic pathways of Sailuotong capsules are NAA metabolism, arginine metabolism, energy metabolism, oxidative stress, etc.

Diagnostic value of acoustic radiation force impulse imaging combined with contrast-enhanced ultrasound in patients with acute cerebral ischemia / 中国中西医结合急救杂志

Objective To investigate the value of acoustic radiation force impulse imaging (ARFI) combined with contrast-enhanced ultrasound (CEUS) in diagnosis of acute cerebral ischemia (ACI) in patients with carotid artery vulnerable plaque (VP). Methods One hundred and ten patients with carotid artery VP diagnosed by routine ultrasound admitted to Renhe Hospital Affiliated to Three Gorges University from October 2017 to January 2019 were selected as subjects, including 59 patients with ACI (ACI group) and 51 patients with non-ACI (non-ACI group) confirmed by magnetic resonance imaging (MRI). All patients underwent routine ultrasound, CEUS and ARFI examinations, the differences in sizes of VP, shear wave velocity (SWV) and enhancement intensity (EI) were compared, and the diagnostic values of SWV and EI were evaluated by the analyses of receiver operating characteristic curve (ROC). Results Two hundred and ten VPs were found in 110 patients, including 112 grade 1 plaques, 53 grade 2 plaques, and 45 grade 3 plaques, compared with grade 1 plaques, the proportion of mixed echo (MP) and strong echo (SP) in grade 2 plaques, and the proportion of all echo types in grade 3 plaques were all significantly different from that of grade 1 plaques (all P < 0.05); compared with grade 1 plaques, the proportion of grade 2 plaques with thickness ≤ 2.0 mm, and the proportion of grade 3 plaques with thickness ≤ 2.0 mm and more than 3.1 mm were all significantly different from that of grade 1 plaques (all P < 0.05). SWV in ACI group was obviously lower than that in non-ACI group (m/s: 1.91±0.54 vs. 2.41±0.57), and EI in ACI group was significantly higher than that in non-ACI group (dB: 3.62±1.13 vs. 2.81±1.05), the difference being statistically significant (both P < 0.05). The area under the ROC curve (AUC) of SWV was 0.681 and the cutoff value was 2.21 m/s, sensitivity, specificity, positive predictive value and negative predictive value were 83.05%, 80.39%, 83.05%, 80.39%; the AUC of EI was 0.638, and the cutoff value was 3.71 dB, sensitivity, specificity, positive predictive value and negative predictive value were 79.66%, 74.51%, 78.33%, 76.00%. AUC of SWV combined with EI was 0.812, sensitivity, specificity, positive predictive value and negative predictive value were 93.22%, 94.12%, 94.83%, 92.31%, significantly higher than those of SWV or EI alone (all P < 0.05). Conclusion In ACI patients, the SWV of VP decreases and EI of VP increases, the detection efficacy of SWV combined with EI for diagnosis of ACI has relatively high clinical value, as the combined diagnostic efficiency is significantly higher than that of either SWV or EI alone.

Lentivirus-Mediated Gene Silencing of Lymphocyte Function-Associated Antigen 1 Inhibits Apoptosis of Hippocampal Neurons in Rats with Acute Cerebral Ischemia After Cerebral Lymphatic Blockage.

BACKGROUND/AIMS: Cerebral ischemia is considered to be the most common cause of stroke with high mortality. It occurs as a result of the damage of the hippocampal neurons with lymphocyte function-associated antigen (LFA)-1 being emphasized to play a role in the biological functions of hippocampal neurons. This study was conducted in order to investigate the effects of specific knockdown of LFA-1 expression by lentivirus had on the apoptosis of the hippocampal neurons, simulated by rat models of acute cerebral ischemia after cerebral lymphatic blockage. METHODS: A total of 60 Wistar rats were selected as subjects, among which 50 were used to establish models of the acute cerebral ischemia after cerebral lymphatic blockage, while the remaining 10 rats were treated with the sham operation. The underlying regulatory mechanisms regarding LFA-1 were analyzed with the treatment of si-LFA-1 and LFA-1 vector in the hippocampal CA1 area of brain tissues isolated from the rats with acute cerebral ischemia. The brain water content, electrolyte content, and blood-brain barrier permeability located in ischemic area of rats were measured. TUNEL staining and immunochemistry methods were employed in order to determine the apoptosis rate and positive levels of LFA-1, MMP-9, and Caspase-3. The mRNA and protein levels of related genes were also detected by means of RT-qPCR and western blot assay. RESULTS: The brain water content, Na+ and Ca+ contents, blood-brain barrier permeability, apoptosis rate, positive levels of LFA-1, MMP-9, and Caspase-3 were decreased, and the K+ content was increased in ischemic tissues treated with si-LFA-1. The mRNA and protein levels of LFA-1, MMP-9, Caspase-3, and Bax had all decreased, while the mRNA and protein levels of Bcl-2 were elevated in the hippocampal CA1 area of rat brain tissues treated with si-LFA-1. These situations could be reversed through the up-regulation of LFA-1. CONCLUSION: In conclusion, LFA-1 gene silencing could improve the acute cerebral ischemia after cerebral lymphatic blockage by inhibiting apoptosis of the hippocampal neurons in rats.

Brain-Derived Glia Maturation Factor ß Participates in Lung Injury Induced by Acute Cerebral Ischemia by Increasing ROS in Endothelial Cells.

Brain damage can cause lung injury. To explore the mechanism underlying the lung injury induced by acute cerebral ischemia (ACI), we established a middle cerebral artery occlusion (MCAO) model in male Sprague-Dawley rats. We focused on glia maturation factor ß (GMFB) based on quantitative analysis of the global rat serum proteome. Polymerase chain reaction, western blotting, and immunofluorescence revealed that GMFB was over-expressed in astrocytes in the brains of rats subjected to MCAO. We cultured rat primary astrocytes and confirmed that GMFB was also up-regulated in primary astrocytes after oxygen-glucose deprivation (OGD). We subjected the primary astrocytes to Gmfb RNA interference before OGD and collected the conditioned medium (CM) after OGD. We then used the CM to culture pulmonary microvascular endothelial cells (PMVECs) acquired in advance and assessed their status. The viability of the PMVECs improved significantly when Gmfb was blocked. Moreover, ELISA assays revealed an elevation in GMFB concentration in the medium after OGD. Cell cultures containing recombinant GMFB showed increased levels of reactive oxygen species and a deterioration in the state of the cells. In conclusion, GMFB is up-regulated in astrocytes after ACI, and brain-derived GMFB damages PMVECs by increasing reactive oxygen species. GMFB might thus be an initiator of the lung injury induced by ACI.

[Scalp-acupuncture Improves Neurological Function by Regulating Expression of NF-κB p 65 mRNA, IκB mRNA and IL-1 ß and TNF-α in Parahippocampal Gyrus of Cerebral Ischemia Rats].

OBJECTIVE: To observe the effect of scalp-acupuncture intervention on the expression of parahippocampal factor-κB p 65 mRNA (NF-κB p 65 mRNA), IκB mRNA, interleukin-1 ß (IL-1 ß) and tumor necrosis factor-α (TNF-α) in rats with cerebral ischemia (CI), so as to investigate its molecular mechanisms underlying improving CI by reducing inflammatory response. METHODS: A total of 64 SD rats were randomized into normal control, model, medication and scalp-acupuncture groups, with 16 rats in each group. The focal CI model was established by middle cerebral artery occlusion (MCAO). Intraperitoneal injection of Pyrrolidine Dithiocarbamate (100 mg•kg－1•d－1) was administrated for rats in the medication group, once a day for 7 days. For rats of the scalp-acupuncture group, the acupuncture needles were rapidly inserted into bilateral Dingnieqianxiexian (MS 6) and Dingniehouxiexian (MS 7), followed by twirling the needles at 200 cycles/min for 1 min, once again every 10 min during 30 min's needle retention. The treatment was conducted once a day for 7 days. The neurologic deficit score (0－5 points, impaired consciousness, death, etc.) and neurological function score (motor, sensory and sensory tests, 0－10 points) were assessed according to Longa's (1989) and Schäbitz's (2004) methods, respectively. The expression levels of NF-κB p 65 mRNA and IκB mRNA in the parahippocampus gyrus tissue were detected by fluorescence quantitative reverse transcription-PCR, and IL-1 ß and TNF-α proteins in the parahippocampus gyrus tissue were detected by immunohistochemistry. RESULTS: After modeling, the neurologic deficit and neurological function scores and the expression levels of NF-κB p 65 mRNA, IL-1 ß and TNF-α in the parahip-pocampus were significantly increased in the model group than in the normal group (P<0.01), while the expression of IκB mRNA was considerably down-regulated (P<0.01). Following treatment intervention, the neurologic deficit and neurological function scores as well as NF-κB p 65 mRNA, and IL-1 ß and TNF-α protein expression were significantly decreased in both scalp-acupuncture and medication groups compared with the model group (P<0.05, P<0.01), and the decreased expression of IκB mRNA was obviously increased (P<0.05)．. CONCLUSION: Scalp-acupuncture can improve neurologic function in cerebral ischemic rats, which is related with its effects in up-regulating the expression of IκB to inhibit the dissociation of NF-κB, then decreasing the expression of IL-1 ß and TNF-α (reducing inflammatory response) in the parahippocampal gyrus tissue.