Neurological Function In Rats Research Articles

Effects of lncRNA gm4419 on rats with hypertensive cerebral atherosclerosis through NF-κB pathway.

To explore the effects of long non-coding ribonucleic acid (lncRNA) Gm4419 on rats with hypertensive cerebral atherosclerosis through the nuclear factor-kappa B (NF-κB) pathway. Healthy male rats were selected and randomly divided into control group, model group (hypertensive cerebral atherosclerosis model), and lncRNA group (hypertensive cerebral atherosclerosis model + lncRNA injection). Neurological deficit scoring criteria, flow cytometry, Western blotting, and staining method were adopted to measure the differences in the neurological function score, NF-κB activity, and chemerin level of rats in the three groups. The neurological scores revealed that the neurological function of rats was not damaged in control group, while it was severely damaged in model group. However, the neurological function of rats was more severely damaged in lncRNA group than that in control group and model group, while the neurological function deficits were slighter in model group. In terms of NF-κB expression activity in mononuclear cells, the serum activity of NF-κB in control group appeared the lowest among the three groups and was significantly higher in lncRNA group than in model group. The serum chemerin level was evidently increased in model group compared with control group, while it was significantly decreased in lncRNA group compared with model group and control group. Moreover, the levels of NF-κB and chemerin were most evidently influenced in lncRNA group. Activating the NF-κB signal, lncRNA Gm4419 promotes the expression of chemerin signal, accelerates the apoptosis of nerve cells, and motivates the deterioration of hypertensive cerebral arteriosclerosis.

Edaravone combined with cinepazide maleate on neurocyte autophagy and neurological function in rats with subarachnoid hemorrhage.

Effects of edaravone combined with cinepazide maleate on neurocyte autophagy and neurological function in rats with subarachnoid hemorrhage were investigated. Eighty SD rats were selected to establish subarachnoid hemorrhage (SAH) rat models, which were divided into sham operation group, SAH group, MCI group and combined group. Hippocampal tissue of each group was taken to observe the number of neurocytes. The expression levels of Beclin-1 and (light chain LC3)-II of rats in each group were detected by ELISA. Pearson's correlation factors were used to analyze the correlation between Beclin-1 and LC3-11, and neurological function tests were carried out on rats of each group 14 and 28 days after administration. The morphological and structural damage of nerve cells in the combined group was further alleviated, and the survival rate of neurons significantly increased at all time points (P<0.05). The expression levels of Beclin-1 and LC3-11 in combined group was significantly higher than those in SAH group and CMI group (P<0.05), and Beclin-1 was positively correlated with LC3-11 (r=0.9454). Longa score of the combined group was significantly lower than that of the other two groups, and muscle strength score was significantly higher than that of the other two groups (P<0.05). Edaravone combined with cinepazide maleate can effectively increase the survival rate of brain cells and promote the volatilization of neurological function in the treatment of hemorrhage in the subretinal space of the omentum, which is worthy of popularization and application.

Platelet-Rich Plasma Combined with Low-Dose Ultrashort Wave Therapy Accelerates Peripheral Nerve Regeneration.

Finding treatments that accelerate peripheral nerve regeneration, prolongation, and functional recovery remains a challenging task. Platelet-rich plasma (PRP) contains numerous growth factors and active proteins, and low-dose ultrashort waves (USWs) stimulate the formation of nerve-nourishing vessels, which are powerful for nerve regeneration. The goal of this study was to evaluate the synergistic effects of serial ultrasound-guided PRP injections combined with low-dose USWs radiation on peripheral nerve regeneration in a crush injury model. Fifty rabbits were equally and randomly divided into normal control, model, USW, PRP, and PRP+USW groups. The neurological function, electrophysiological recovery, and histological and morphological evaluation of regenerated nerves, as well as a targeted muscle recovery assessment, were performed to investigate the regenerative effect of PRP combined with USW therapy. Our results showed that the PRP+USW group had the better early axonal regeneration and displayed the earliest positive compound muscle action potential among the treatment groups. At postintervention week 12, a histological evaluation showed similar expression of the S-100 protein in the PRP+USW and normal control groups. Moreover, the morphological assessment revealed a significant increase in the myelinated nerve fiber density and diameter and myelin sheath thickness compared with the USW and PRP groups. The morphometry of the target muscles indicated the lowest reduction in the percent volume in the PRP+USW group, and an ultrasound examination of the targeted muscle showed the best improvement in stiffness and perfusion parameters at 12 weeks after crush injury. Thus, serial ultrasound-guided PRP injections combined with low-dose USW radiation exert a synergistic effect on accelerating functional axon recovery and decreasing atrophy of the target muscles in a crush injury model. Impact Statement This research describes that the application of platelet-rich plasma combined with low-dose ultrashort waves treatment exert a synergistic effect on accelerating peripheral nerve regeneration. With the extensive use of platelet-rich plasma and physical factors in regenerative medicine or clinical rehabilitation medicine, our findings may help establish effective strategies for repairing peripheral nerve injury.

Protective effect of propofol on neurological function in rats after traumatic brain injury and related mechanism

Objective To investigate the protective effect of propofol on neurological function in rats after traumatic brain injury (TBI) and its possible mechanism. Methods A total of 96 SD rats were randomly divided into sham operation group, sham operation+ propofol group, TBI group and TBI + propofol group, with 24 rats in each group. The TBI model was prepared by modified Feeney method. The sham operation+ propofol group and the TBI+ propofol group were given 50 mg/kg of propofol once daily. The sham operation group and the TBI group were injected with the same amount of normal saline. Modified neurobehavioral functional scores (mNSS) were evaluated at 1, 3, 7 and 14 days after injury; dry-wet specific gravity method was used to detect brain water content in injured area; TUNEL staining was used to detect neuronal apoptosis; chemiluminescence was used to detect activity of Oxygen cluster (ROS) content; Western blot was used to determine the expressions of inositol requirement enzyme 1 (IRE-1), enhancer binding protein homolog protein (CHOP), heme oxygenase 1 (HO-1), quinone oxidoreductase 1 (NQO1) and nuclear factor E2 related factor 2 (Nrf2) protein. Results Compared with the sham operation group and the sham operation + propofol group, the mNSS, brain tissue water content, apoptosis number and ROS increased at 1, 3, 7 and 14 days after TBI in the TBI group and TBI + propofol group (P<0.05). Compared with TBI group, mNSS in TBI+ propofol group decreased significantly [(9.3±1.4)points ∶(10.9±1.2)points] 7 days after injury (P<0.05); the brain tissue water content decreased significantly [(81.0±0.8)%∶(82.1±0.8)%] 3 days after injury (P<0.05); the number of apoptotic cells decreased significantly 7 days after injury[(14.1±1.4)%∶(15.6±1.6)%], with the most significant decrease at 14 days after injury [( 10.4±1.5)%∶(13.2±1.4)% (P<0.05); and ROS decreased significantly 7 days after injury [(61.5±4.0)RFU∶(77.3±5.5)RFU](P<0.05). Compared with the sham operation group and the sham operation+ propofol group, the expressions of IRE-1 and CHOP were significantly up-regulated in the TBI group and the TBI+ propofol group (P<0.05); the expressions of HO-1, NQO1 and Nrf2 in the TBI group were significantly decreased (P<0.05); the expressions of HO-1 and NQO1 in TBI+ propofol group were increased (P<0.05) while the expression of Nrf2 were decreased slightly (P<0.05). Compared with the TBI group, the expressions of IRE-1 and CHOP in TBI+ propofol group were decreased (P<0.05), while the expressions of HO-1, NQO1 and Nrf2 were significantly increased (P<0.05). Conclusion After TBI in rats, propofol can reduce oxidative stress by activating the Nrf2-antioxidant element (ARE) pathway, reduce brain edema, and inhibit neuronal apoptosis, thus playing a neuro-protective role. Key words: Brain injuries; Oxidative stress; Nuclear factor-E2-related factor 2

P5746Human induced pluripotent stem cell-derived mesenchymal stem cell therapy effectively reduced brain infarct volume and preserved neurological function in rat after acute intracranial hemorrhage

Abstract Intracerebral hemorrhage (ICH) causes 10%-20% of all strokes and results in higher morbidity compared to other subtypes of cerebral stroke. Although early surgical intervention can clear the expanding hematoma, clinical outcomes following ICH have not significantly improved over the decades. Since ICH elicits neuroinflammation to exacerbate brain edema, damage the blood-brain barrier (BBB), lead to secondary neuronal injury, anti-inflammation may be a critical therapeutic strategy. Mesenchymal stem cell (MSC) therapy processes anti-inflammatory, immunomodulatory and tissue regenerative properties, suggesting that MSC therapy could be an effective therapy for ICH. Therefore, this study tested the hypothesis that human induced pluripotent stem cell-derived mesenchymal stem cell (iPSC-MSC) therapy could effectively reduce brain-infract volume (BIV) and improve neurological function in rat after acute ICH induced by a weight-drop device. Adult-male SD rats (n=40) were equally divided into group 1 (sham-operated control), group 2 (ICH), group 3 (ICH + hyaluronic acid (HA)/intracranial injection/3h after ICH), group 4 [ICH + HA + iPSC-MSC (1.2x106 cells/intracranial injection/3h after ICH)] and euthanized by day 28 after ICH procedure. In vitro study showed that hemorrhagic-brain tissue augmented protein expressions of inflammation (HMGB1/MyD88/TLR-4/TLR-2/NF-κB/TNF-α/iNOS/IL-1β) in cultured neurons that were significantly inhibited by iPSC-MSC treatment (all p&lt;0.001). By days 7/14 after ICH procedure, circulating inflammatory levels of TNF-α/IL-6/MPO expressed were lowest in group 1, highest in group 2 and significantly lower in group 4 than in group 3 (all p&lt;0.0001). By day 14 after ICH procedure, neurological function and BIV expressed an opposite pattern, whereas protein expressions of inflammation (HMGB1/MyD88/TLR-4/TLR-2/NF-κB/I-kB/TNF-α/iNOS/IL-1β/MMP-9), oxidative stress (NOX-1/NOX-2/oxidized protein) and apoptosis (mitochondrial-Bax/cleaved-caspase-2/PARP) in brain exhibited an identical pattern to circulating inflammation among the four groups (all p&lt;0.001). Microscopy demonstrated that the number of vascular remodeling/GFAP+/53BP1+/γ-H2AX+ cells displayed an identical pattern of inflammation, whereas the NeuN+ cells displayed an opposite pattern of inflammation among the four groups (all p&lt;0.001). In conclusion, iPSC-MSC therapy markedly reduced BIV and preserved neurological function mainly by inhibiting inflammatory/oxidative-stress generation. Acknowledgement/Funding Kaohsiung Chang Gung Memorial Hospital, Taiwan Society of Stem Cell Research

Cyclosporine A Plus Ischemic Postconditioning Improves Neurological Function in Rats After Cardiac Resuscitation.

Attenuation of neuronal apoptosis helps maintain neurological function in patients after cardiac arrest. After ischemia-reperfusion, both cyclosporin A (CsA) and ischemic postconditioning independently protect mitochondria and thus reduce nerve injury. This study employed a rat model to evaluate the neuroprotective effect of combining ischemic postconditioning with CsA after cardiopulmonary resuscitation (CPR). Rats were apportioned equally to model control, postconditioned, CsA-treated, or CsA + postconditioned groups. Asphyxial cardiac arrest was imposed using modified Utstein-style guidelines. In the appropriate groups, postconditioning was implemented by ischemia and reperfusion (clamping and loosening the left femoral artery); CsA treatment was delivered with a single intravenous dose. Neurological deficits were scored at different times after CPR. Histological evaluation and electron microscopy were used to evaluate tissue damage, and TUNEL and flow cytometry were used to measure the apoptotic rate of hippocampal neurons and size of the mitochondrial permeability transition pore (mPTP) opening. The apoptotic rate was significantly lower in the postconditioned and CsA-treated groups compared with the model control and lowest in the CsA + postconditioned group. By histological evaluation and electron microscopy, the least damage was observed in the CsA + postconditioned group. The neurological deficit score of the CsA + postconditioned group was significantly higher than that of the CsA-treated group, but the size of the mPTP openings of these two groups was comparable. Ischemic postconditioning combined with CsA exerted a better neuroprotective effect after CPR than did either postconditioning or CsA alone. Inhibiting the opening of the mPTP is not the only neuroprotective mechanism.

Granulocyte colony-stimulating factor and stromal cell-derived factor-1 combination therapy: A more effective treatment for cerebral ischemic stroke.

Drugs that promote angiogenesis include statins, recombinant human granulocyte colony-stimulating factor, and stromal cell-derived factor-1. Low doses of atorvastatin could significantly increase the vascular expressions of endothelial growth factor, and the number of peripheral blood endothelial progenitor cells (EPCs), thus improving angiogenesis and local blood flow. G-CSF is an EPC-mobilization agent used in ischemia studies for targeting angiogenesis after cerebral ischemia via EPCs. In previous clinical trials, consistent conclusions have not been reached about the effectiveness of G-CSF on ischemic stroke. Therefore, the therapeutic effect of G-CSF and its combination with other medicines need further experimental verification. It is known that atorvastatin, rhG-CSF, and SDF-1 are considered the most promising neuroprotective candidates, but a comprehensive comparison of their effects is lacking. To compare the effects of atorvastatin, stromal cell-derived factor-1, and recombinant human granulocyte colony-stimulating factor on ischemic stroke. Adult male Sprague-Dawley rats were randomly allocated to three groups: normal, sham-operated, and middle cerebral artery occlusion operated. Middle cerebral artery occlusion operated rats were further allocated into saline, atorvastatin, recombinant human granulocyte colony-stimulating factor, and recombinant human granulocyte colony-stimulating factor + stromal cell-derived factor-1 groups. Neurological function evaluation, cerebral infarction and the blood-brain barrier integrity analysis, identification of angiogenic factors, assessment of angiogenesis, expression of growth-associated protein-43, neuroglobin, glial cell-derived neurotrophic factor, and cleaved caspase 3, were performed. Compared with atorvastatin or recombinant human granulocyte colony-stimulating factor alone, recombinant human granulocyte colony-stimulating factor + stromal cell-derived factor-1 treatment improved neurological performance, reduced cerebral infarction and blood-brain barrier disruption after stroke, and increased the content of stromal cell-derived factor-1, vascular endothelial growth factor, monocyte chemotactic protein 1, and basic fibroblast growth factor in peripheral blood. In addition, recombinant human granulocyte colony-stimulating factor + stromal cell-derived factor-1 promoted greater angiogenesis than atorvastatin or recombinant human granulocyte colony-stimulating factor alone and increased the expression of growth-associated protein-43, neuroglobin, and glial cell-derived neurotrophic factor, while decreasing the levels of cleaved caspase 3 in the brain after ischemic stroke. Combination therapy with recombinant human granulocyte colony-stimulating factor and stromal cell-derived factor-1 is more effective than atorvastatin or recombinant human granulocyte colony-stimulating factor alone in protecting against stroke-induced damage and could be an optimal therapeutic strategy for stroke.

P.279Pragmatic cognitive and psychosocial health monitoring in clinical practice in Duchenne muscular dystrophy

We performed a prospective, cross-sectional cognitive assessment in subjects with Duchenne muscular dystrophy (DMD). Thirty subjects (29 boys and 1 girl) with out-of-frame mutations in dystrophin (DMD) gene were assessed using the National institutes of health toolbox cognition battery (NIHTB-CB). The NIH toolbox is a scientifically rigorous, multi-dimensional measure designed to bring uniformity to evaluation of neurological function across the life span. The NIHTB-CB tests have previously been evaluated for their reliability, validity, and other psychometric properties, including among individuals with several different neurodevelopmental disorders. A caregiver completed the Behavior Rating Inventory of Executive Functioning (BRIEF), a standardized rating scale of executive functioning, for their child. Overall, the children with DMD scored one standard deviation (SD) below age-corrected norms on the NIHTB-CB total cognition score. They scored 1.5 SD below age-corrected norms in fluid cognition, which evaluates the cognitive domains of executive function, working memory, episodic memory, attention, and processing speed. In crystallized cognition score, which evaluates vocabulary and reading, their performance was consistent with age expectations (within 1 SD below age-corrected norms). Children with DMD had higher T-scores in several domains of BRIEF, demonstrating greater difficulty in executive functioning. The NIHTB-CB sensitively identifies core deficits in the specific domains of cognition in DMD, and can be easily incorporated in a clinic setting to assess cognitive needs and psychosocial health in DMD.

SOD3 overexpression alleviates cerebral ischemia-reperfusion injury in rats.

BackgroundIschemic stroke is a deadly disease that poses a serious threat to human life. Superoxide dismutase 3 (SOD3, ECSOD) is the main antioxidant enzyme that removes superoxide anions from cells. This study aimed to investigate the effect of SOD3 overexpression on cerebral ischemia‐reperfusion injury in rats.MethodsGV230‐EGFP‐ECSOD, the recombinant SOD3‐overexpressed vector, was constructed by genetic engineering technology, and mesenchymal stem cells (MSCs) were infected with lentiviral packaging. In animal experiment, cerebral ischemia‐reperfusion injury model rats were successfully established. ECSOD‐MSCs are the MSCs that successfully transfected with SOD3 overexpression vector. The animals were injected with ECSOD‐MSCs (ECSOD‐MSC group), normal MSCs (MSCs group), PBS (PBS group), and not do any processing (Model group) via the tail vein. Then MRI was used to detect the infarct volume of rats, modified Neurological Severity Scores (mNSS), and immunohistochemistry were used to evaluate the expression of neurological function and apoptosis‐related genes in rats.ResultsWestern blot analysis revealed that the SOD3 was highly expressed in MSCs. Animal experiments showed that the transplantation of ECSOD‐MSCs significantly reduced the infarct volume of ischemic stroke rats (p < 0.05), significantly improved neurological function in rats (p < 0.05), and found proapoptotic gene, Bax, expression was significantly decreased (p < 0.05), the expression of anti‐apoptotic gene, Bcl‐2, was significantly increased (p < 0.05). The highly expressed SOD3 has no correction with brain infarct volume, and the highly expressed SOD3 has a positive correlation with cell apoptosis. It is speculated that overexpression of SOD3 affects the expression of Bax and Bcl‐2, and improves apoptosis to alleviate ischemic stroke.ConclusionOur results indicated that MSCs transfected with SOD3 can effectively alleviate cerebral ischemia‐reperfusion injury in rats.

Effects of ISO-Alpha-Acids on the Hematoma Volume, Inflammation and Neurological Functions in Intracerebral Hemorrhage Rats and its Potential Mechanism

Abstract INTRODUCTION ISO-alpha-acids (IAA) was reported to be useful for the prevention of dementia as a PPAR-gamma agonist in rats, but the application of IAA in intracerebral hemorrhage (ICH) was still limited. METHODS ICH models were built in Vivo and 90 SD rats were randomized to 5 groups: ICH + IAA intraperitoneal injection (IAA), ICH + normal saline intraperitoneal injection (control), ICH without any intraperitoneal injections (vehicle), Sham, and Mock. Neurological functions of rats, hematoma volume, and brain water content were evaluated 1-d, 3-d, and 7-d postoperation. Microglia of SD rats were sub cultured in Vitro. According to different interventions, 4 groups were set, including IAA, PPAR-gamma antagonist, normal saline (control), and normal cultured (vehicle). Western blotting, immunofluorescence, flow cytometry, and RT-PCR were used to detect the expression of molecular markers, proteins and mRNAs. RESULTS In ICH rats, intraperitoneal injection of IAA was able to promote hematoma clearance, to relieve brain water content and improve neurological functions. Detected by western blot, immunofluorescence, flow cytometry, and reverse transcription polymerase chain reaction (RT-PCR), IAA was able to significantly increase the expression of CD36, CD11b, and CD206, and inhibit the expression of CD80, CD86, TLR4, NF-kappaB, TNF-alpha, and IL-1beta comparing with other groups (P &lt; .05) in Vivo and Vitro. IAA was able to significantly increase both in CD11b and CD206 double-positive anti-inflammatory type microglia and in microglia phagocytosis in the brain (P &lt; .05). CONCLUSION IAA induces microglia to M2 type, characterized by phagocytosis and anti-inflammation, via activating PPAR-gamma signaling pathway in Vivo and Vitro. In ICH rats, administration of IAA is able to facilitate intracerebral hematoma absorption, alleviate brain edema, suppress inflammatory reactions, protect neurological functions, and finally improve rat's outcome.

Effect of miR-124 on neuronal apoptosis in rats with cerebral infarction through Wnt/β-catenin signaling pathway.

The aim of this study was to observe the influence of micro ribonucleic acid (miR)-124 on neuronal apoptosis in rats with cerebral infarction (CI), and to further investigate the underlying mechanism of miR-124 in CI occurrence and development. A total of 60 adult male Wistar rats were randomly divided into the Sham group, the CI group and the CI + miR-124 mimics group using a random number table. The focal CI model was established using the suture-occluded method. After successful modeling, miR-124 mimics were stereotactically injected into the lateral ventricle of rats. 24 h after operation, the neurological function of rats in each group was scored using modified neurological severity score (mNSS). Meanwhile, the infarction area of brain tissues was evaluated by the triphenyl tetrazolium chloride (TTC) method. The protein expression levels of apoptosis-related genes, including B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X protein (Bax), C-Caspase and T-Caspase, were detected via Western blotting. The expression and location of caspase-3 in brain tissues were detected via immunofluorescence staining. Moreover, the level of apoptosis in each group was detected via terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. In addition, the expression levels of the Wnt/β-catenin signaling pathway-related proteins were detected via Western blotting. Polymerase Chain Reaction (PCR) results revealed that the expression level of miR-124 in the CI group was significantly decreased when compared with that of the Sham group (p<0.05). The mNSS and TTC staining results manifested that injection of miR-124 mimics could significantly reduce the CI-induced neurological deficits and CI area (p<0.05). At the same time, the levels of Bax and C-caspase/T-Caspase were significantly decreased, whereas the expression of Bcl-2 was remarkably increased after the injection of miR-124 mimics (p<0.05). Besides, the number of apoptotic cells in the CI + miR-124 mimic group was remarkably decreased (p<0.05). In addition, miR-124 mimics significantly activated the expression levels of Wnt and β-catenin (p<0.05). The inhibitory effect of miR-124 on neuronal apoptosis in CI rats is probably related to the activation of the Wnt/β-catenin signaling pathway. Furthermore, miR-124 is expected to be a target drug for the clinical treatment of CI.

14-3-3ζ protein protects against brain ischemia/reperfusion injury and induces BDNF transcription after MCAO in rat.

Brain ischemia is a leading cause of death and disability worldwide that occurs when blood supply of the brain is disrupted. Brain-derived neurotrophic factor (BDNF) is a protective factor in neurodegenerative conditions. Nevertheless, there are some problems when exogenous BDNF is to be used in the clinic. 14-3-3ζ is a pro-survival highly-expressed protein in the brain that protects neurons against death. This study evaluates 14-3-3ζ effects on BDNF transcription at early time point after ischemia and its possible protective effects against ischemia damage. Human 14-3-3ζ protein was purified after expression. Rats were assigned into four groups, including sham, ischemia, and two treatment groups. Stereotaxic cannula implantation was carried out in the right cerebral ventricle. After one week, rats underwent middle cerebral artery occlusion (MCAO) surgery and received 14-3-3ζ (produced in our laboratory or standard form as control) in the middle of ischemia time. At 6 h of reperfusion after ischemia, brain parts containing the hippocampus, the cortex, the piriform cortex-amygdala and the striatum were collected for real time PCR analysis. At 24 h of reperfusion after ischemia, neurological function evaluation and infarction volume measurement were performed. The present study showed that 14-3-3ζ could up-regulate BDNF mRNA at early time point after ischemia in the hippocampus, in the cortex and in the piriform cortex-amygdala and could also improve neurological outcome and reduce infarct volume. It seems that 14-3-3ζ could be a candidate factor for increasing endogenous BDNF in the brain and a potential therapeutic factor against brain ischemia.

Hypothermic properties of dexmedetomidine provide neuroprotection in rats following cerebral ischemia-reperfusion injury.

Dexmedetomidine (Dex) is a sedative and analgesic agent that is widely administered to patients admitted to the intensive care unit, and has been demonstrated to result in hypothermia. Many patients have been revealed to benefit from therapeutic hypothermia, which can mitigate cerebral ischemia/reperfusion (I/R) injury following successful cardiopulmonary resuscitation. However, studies investigating the efficacy of Dex in I/R treatment is lacking. The present study aimed to investigate the efficacy of Dex in mitigating neuronal damage, and to determine the possible mechanism of its effects in a rat model of cardiac arrest (CA). CA was induced in Sprague-Dawley rats by asphyxiation for 5 min. Following successful resuscitation, the surviving rats were randomly divided into two treatment groups; one group was intraperitoneally administered with Dex (D group), whereas the control group was treated with normal saline (N group). Critical parameters, including core temperature and blood pressure were monitored following return of spontaneous circulation (ROSC). Arterial blood samples were collected at 10 min after surgery (baseline) 30 and 120 min post-ROSC; and neurological deficit scores (NDS) of the rats were taken 12 or 24 h after ROSC prior to euthanasia. The hippocampal tissue was then removed for analysis by histology, electron microscopy and western blotting. Rats in the D group exhibited a lower core temperature and higher NDS scores compared with the N group (P<0.05). In addition, Dex injection resulted in reduced expression of apoptotic and autophagy-associated factors in the hippocampus (P<0.05). Dex treatment induced hypothermia and improved neurological function in rats after ROSC following resuscitation from CA by inhibiting neuronal apoptosis and reducing autophagy, which suggested that Dex may be a potential therapy option for patients with CA.

Mitoquinone attenuates blood-brain barrier disruption through Nrf2/PHB2/OPA1 pathway after subarachnoid hemorrhage in rats

Background and purposeMitochondrial dysfunction is involved in the mechanism of early brain injury (EBI) following subarachnoid hemorrhage (SAH). Blood-brain barrier disruption is a devastating outcome in the early stage of SAH. In this study, we aimed to investigate the role of a mitochondria-related drug Mitoquinone (MitoQ) in blood-brain barrier disruption after SAH in rats. MethodsA total of 181 male Sprague–Dawley SAH rats with the endovascular perforation model were utilized. Intraperitoneal MitoQ was given 1 h (h) post-SAH. Cerebroventricular ML385, an inhibitor of NF-E2-related factor 2 (Nrf2) and Small interfering ribonucleic acid (siRNA) for Prohibitin 2 (PHB2) were injected respectively 24 h and 48 h before SAH. Neurological function evaluation was performed before sacrifice. SAH grade was measured during the sacrifice of each animal. Brain water content was performed at 24 h. Co-immunoprecipitation was used to demonstrate the relationship of proteins Nrf2 and PHB2. Mitochondrial and cytoplasmic fractions were gathered using mitochondria isolation kits. Pathway related proteins were investigated with Western blot and immunofluorescence staining. Transmission electron microscopy was performed for mitochondrial morphology. ResultsExpression of Nrf2 levels peaked at the 3 h time point following SAH and then decreased to normal levels at 24 h, while PHB2 and Optic Atrophy 1 (OPA1) decreased at 24 h and 72 h after SAH compared with the Sham group. MitoQ treatment attenuated neurological deficits and brain edema, thereby resulting in a decreased expression of Albumin, while an increase of Nrf2, PHB2, OPA1 and Claudin-5 proteins compared with SAH + vehicle group. With co-immunoprecipitation, Nrf2 and PHB2 were further demonstrated to show their interaction. And MitoQ administration lead to more binding of the two proteins. ML385 abolished the effects of MitoQ on neurobehavior and protein levels post-SAH. Similarly, PHB2 siRNA reversed the neuroprotection of MitoQ administration with the decreased expression of PHB2 and OPA1 after SAH. Further, MitoQ treatment improved mitochondrial morphology after SAH with an increase of PHB2 and OPA1 in mitochondrial extraction. ConclusionsMitoQ attenuates blood-brain barrier disruption via Nrf2/PHB2/OPA1 pathway after SAH in rats. MitoQ may serve as a potential therapeutic strategy for SAH patients.

Repetitive Transcranial Magnetic Stimulation Improves Neurological Function after Cerebral Ischemia in Rats by Increasing CREB-Regulated TrkB &lt;i&gt;via&lt;/i&gt; Activation of cAMP/PKA and Ca &lt;sup&gt;2+&lt;/sup&gt;/CaMKIV Signaling Pathways

Cerebral ischemia is the most prevalent form of clinical stroke. Repetitive transcranial magnetic stimulation (rTMS) can modulate excitability of the cerebral cortex, and this effect is maintained after the stimulation is terminated. However, the underlying mechanisms of rTMS in cerebral ischemia remain unclear. Herein, we identified the effect of rTMS on cerebral ischemia. The in vitro model was established using primary cultured neurons under the condition of oxygen-glucose deprivation (OGD), followed by 1 Hz or 10 Hz rTMS treatment. The levels of CREB, PKA and CaMKIV were depleted in neurons to explore the underlying regulation mechanisms of TrkB by rTMS via CREB. Rat models with cerebral ischemia were established by middle cerebral artery occlusion (MCAO) and stimulated with 1 Hz or 10 Hz rTMS to investigate the effect of rTMS on neurobehavior, CREB expression, cAMP/PKA and Ca2+/CaMKIV pathways. rTMS was observed to promote the nerve recovery ability in rats with cerebral ischemia, which was accompanied by high expression of TrkB. Besides, in OGD-treated neurons, rTMS activated CREB by upregulating cAMP/PKA and Ca2+/CaMKIV pathways. Moreover, rTMS induced the activation of CREB to upregulate TrkB. In MACO rats, rTMS promoted CREB expression and the content of CREB, cAMP and PKA phosphorylation, increased the CaMKIV phosphorylation, and promoted the interaction between CREB and TrkB. Taken together, rTMS promoted CREB expression and up-regulated TrkB to improve the neurological function in rats with cerebral ischemia by activating cAMP/PKA and Ca2+/CaMKIV pathways, which could be of great significance for cerebral ischemia therapy. Funding Statement: The authors declare: None. Declaration of Interests: The authors declare: None. Ethics Approval Statement: The animal experiment was performed in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. The protocol was approved by the Animal Ethics Committee of North China University of Science and Technology Affiliated Hospital.

Neurological functional evaluation based on accurate motions in big animals with traumatic brain injury.

An accurate and effective neurological evaluation is indispensable in the treatment and rehabilitation of traumatic brain injury. However, most of the existing evaluation methods in basic research and clinical practice are not objective or intuitive for assessing the neurological function of big animals, and are also difficult to use to qualify the extent of damage and recovery. In the present study, we established a big animal model of traumatic brain injury by impacting the cortical motor region of beagles. At 2 weeks after successful modeling, we detected neurological deficiencies in the animal model using a series of techniques, including three-dimensional motion capture, electromyogram and ground reaction force. These novel technologies may play an increasingly important role in the field of traumatic brain injury diagnosis and rehabilitation in the future. The experimental protocol was approved by the Animal Care and Use Committee of Logistics University of People's Armed Police Force (approval No. 2017-0006.2).

Application of 18F-FDG microPET/CT in the screening of cerebral ischemia reperfusion models

Objective To investigate the feasibility of 18F-fluorodeoxyglucose (FDG) microPET/CT in the screening of cerebral ischemia reperfusion (CIR) models. Methods The suture-occluded method was used to establish CIR rat models with reversible middle cerebral artery embolism. After that only awake rats whose Zea-Longa scores were 1-4 were selected for the following experiments, and 18 male SD rats were selected. Garcia scale with 18 points was used to evaluate the neurological function of rats at 2 and 24 h post-operation. At the same time points, 18F-FDG was injected into caudal vein after anesthesia and microPET/CT scan was conducted at 40 min post-injection. Visual and semi-quantitative analyses were adopted to analyze the images. The autopsy and HE staining were performed on accidentally dead rats. The other alive rats were sacrificed after microPET/CT scan at 24 h post-operation, and their brain tissues were taken out quickly to detect the infarction by triphenyl tetrazolium chloride (TTC) staining. The pathological results were taken as the gold criteria. Fisher exact test was used to compare the difference of accuracy for diagnosing CIR models between neurological function score (NFS) and microPET/CT. Results According to the pathology, there were 11 CIR models, 4 with subarachnoid hemorrhage (SAH), 3 with SAH and cerebral hemorrhage. Between 8-12 h post-operation, 4 rats died accidentally. At 2 h post-operation, the diagnostic accuracies of NFS and microPET/CT were 11/18 and 15/18 (P 0.05). Conclusion 18F-FDG microPET/CT is better than NFS in screening CIR models in early stage. Key words: Brain ischemia; Reperfusion; Positron-emission tomography; Tomography, X-ray computed; Deoxyglucose; Rats

Jobelyn® ameliorates neurological deficits in rats with ischemic stroke through inhibition of release of pro-inflammatory cytokines and NF-κB signaling pathway

The effects of Jobelyn® (JB) on neurological deficits and biochemical alterations associated with ischemic stroke induced by bilateral common carotid artery occlusion (BCCAO) in rats were investigated in this study. Male Wistar rats were divided into five groups (n = 8): group 1 served as Sham control; group 2, which served as negative control received normal saline while groups 3–5 were given JB (25, 50 and 100 mg/kg, p.o) daily for 28 days. Then, rats in groups 2–5 were subjected to BCCAO for 30 min and reperfusion afterwards. Neurological deficits were assessed 3 h post-reperfusion using a 9-point neurological scoring scale. The levels of biomarkers of oxidative stress and pro-inflammatory cytokines (tumour necrotic factor-α and interleukin-6), expressions of immunopositive cells of nuclear factor-kappa B (NF-κB) and acetyl-cholinesterase (AChE) activity were determined in brain tissues. Histology of the striatum, prefrontal cortex (PFC) and hippocampus (CA1) was also evaluated. JB improved BCCAO-induced neurological deficits and attenuated increased oxidative stress and AChE activity in rats subjected to BCCAO (p < 0.05). Increased brain levels of tumour necrotic factor-α and interleukin-6 as well as expressions of immunopositive cells of NF-κB were decreased by JB. JB reduced brain damage and also increased population of viable neurons in the striatum, PFC and hippocampus of ischemic stroke rats. These findings suggest that the positive effect of JB on neurological function in rats with ischemic stroke may be related to inhibition of oxidative stress, release of pro-inflammatory cytokines and expressions of immunopositive cells of NF-κB.

Application of microprobe vascular Doppler in STA-MCA bypass intraoperation

Objective To assess the reliability and efficacy of microprobe vascular Doppler(MVD) evaluating the patency of anastomotic vessels in patients with unilateral symptomatic carotid artery occlusion(SICAO) undergoing superficial temporal artery to middle cerebral artery bypass surgery(STA-MCA bypass). Methods Total 50 cases of SICAO patients from July 2012 to May 2016 at Department of Neurosurgery, the First Affiliated Hospital of Soochow University, confirmed by DSA and CT perfusion were treated with STA-MCA bypass surgery. The hemodynamics parameters of M4 segment of middle cerebral artery(MCA) and superficial temporal artery (STA) were evaluated by MVD intraoperation. Postoperative CTP and DSA examinations were performed to evaluate the perfusion of the ischemic area and the patency of anastomotic vessels. Recurrent stroke was recorded 6-12 months after operation and neurological recovery was assessed by modified Rankin scale(mRS). Results (1)Intraoperation: MVD revealed the blood flow velocity of MCA M4 segment and the pulsatility index(PI) increased(all P<0.001) and the flow direction of M4 proximal was reversed. The blood flow velocity of STA increased but the PI sharply decreased(all P<0.001) and it appeared intracranial arterial spectrum . (2)Seven days after operation: Carotid Doppler ultrasonography (CDU) revealed the diameter and the blood flow volume of STA anterior to the tragus obviously increased (P<0.05). CTP revealed the rCBF of basal ganglia level increased, the rMTT and rTTP decreased and the difference were statistically significant(all P<0.05). (3)Six months after operation: DSA revealed the patency of anastomotic vessels access 96%(48/50). (4)Evaluation of recurrent stroke and neurological function: The postoperative mRS score decreased or unchanged in 96%(48/50)of the patients. No recurrent stroke was found in 48 patients with the patency of anastomotic vessels. Conclusions MVD can evaluate hemodynamic changes in STA-MCA bypass surgery in a simple, no-invasive and dynamic way. When STA showing the Intracranial arterial spectrum is a reliable index to judge the patency of anastomotic vessels. Key words: Ultrasonography, Doppler; Carotid artery, internal; Hemodynamics; Microsurgery

Effects of erythropoietin administrated by intranasal on nerve function and CD31 in cerebral ischemia reperfusion rats

Objective To explore the protective effect of erythropoietin(EPO) administrated by intranasal on cerebral ischemia reperfusion in rats with acute cerebral infarction reperfusion. Methods Total of 100 SD rats were divided into model control group, sham operation group, intraperitoneal administration group (IPEPO group), nasal saline group (INNS group) group, and nasal drug delivery group (INEPO group) with 20 in each group.The middle cerebral artery occlusion model of rat was established by thread embolism method and the NSS method was used to evaluate the neural behavior of rats.The expression of EPO in peripheral blood, cerebrospinal fluid and brain regions of rats were detected by Elisa.The vascular endothelial growth factor(VEGF) in brain was detected by immunofluorescence and then the density of newborn blood vessels in the brain was measured. Results Fifteen days after the operation, the NSS score of INEPO group(3.80±1.61) was significantly lower than that of IPEPO group (11.53±2.11), and the difference was statistically significant(P<0.01). And the levels of EPO in blood, cerebrospinal fluid and different brain regions of rats in INEPO group were higher than that of INNS group(all P<0.01). Compared with IPEPO group, the level of EPO cerebrospinal fluid and different brain regions of rats in INEPO group increased obviously, the difference was statistically significant (all P<0.01), and the EPO concentration of the olfactory bulb was the most obvious(INEPO group: (1 456.90±128.22)pg/ml, IPEPO group: (426.11±36.68)pg/ml, P<0.01). Seventy-two hours after operation, the expression of CD31 in ischemic penumbra of rats of model control group (18.21±3.45), INNS group(18.54±2.58), IPEPO group(27.01±2.13)and INEPO group(35.52±2.79)was increased compared with sham operation group (5.14±1.28), and the difference was statistically significant (all P<0.05). The expression of CD31 in IPEPO group and INEPO group was significantly higher than that in INNS group (P<0.05). In INEPO group, the expression of CD31 increased significantly compared with that of IPEPO group (P<0.05). Conclusion Nasal administration of EPO can effectively improve the neurological function of rats with ischemia-reperfusion, and increase the expression of CD31 in the brain tissue of rats.The effect of nasal administration is better than that of intraperitoneal administration. Key words: Nasal administration; Erythropoietin; Ischemia reperfusion; Cerebral infarction; Angiogenesis; Rat