Treatment Of Middle Cerebral Artery Occlusion Research Articles

BONE MARROW MESENCHYMAL STROMAL CELL-DERIVED EXOSOMAL NRF2 AMELIORATES CEREBRAL ISCHEMIA-REPERFUSION INJURY BY TRANSCRIPTIONALLY ACTIVATING LIN28A.

Background: Cerebral ischemia-reperfusion (I/R) injury (CIRI) have severe consequences on brain function, and the exciting evidence has revealed protective role of acyl-CoA synthetase long chain family member 4 (Lin28a) against cerebral ischemia-reperfusion injury. The present work aims to reveal its molecular mechanism in regulating CIRI, with the hope of providing a therapeutic method for cerebral I/R injury. We hypothesized that the exosomal nuclear factor erythroid 2-related factor 2 (NRF2) derived from bone marrow mesenchymal stromal cells could transcriptionally activate Lin28a and thereby alleviate cerebral ischemia-reperfusion injury. This hypothesis was validated in the present work. Methods: Middle cerebral artery occlusion (MCAO) model was established using C57BL/6J mice, and the neurological deficit, infarct volume, and brain water content were assessed to evaluate neuron injury. Human glioblastoma cells (A172) were subjected to oxygen-glucose deprivation and reoxygenation (OGD/R) treatment to mimic a cerebral I/R injury cell model. Exosome isolation reagent was used to isolate exosomes from cell supernatant of bone marrow mesenchymal stromal cells through sequential centrifugation and filtration steps. mRNA expression level of Lin28a was detected by quantitative real-time polymerase chain reaction. Protein expression was analyzed by western blotting assay. TUNEL cell apoptosis detection kit was used to analyze cell apoptosis in brain tissues. Enzyme-linked immunosorbent assays and commercial kits were used to detect levels of inflammatory markers and oxidative stress markers. Ferrous Iron Colorimetric Assay Kit and Fe 2+ colorimetric assay kit were used to analyze Fe 2+ level. The association of Lin28a and NRF2 was identified by chromatin immunoprecipitation assay and dual-luciferase reporter assay. Results: The treatment of MCAO substantially augmented infarct volume in mice, impaired neurological function, and elevated brain water content. Lin28a was lowly expressed in brain tissues of mice with CIRI, and its overexpression protected against cerebral I/R injury of MCAO mice. Moreover, Lin28a overexpression protected A172 cells against OGD/R treatment-induced injury. Additionally, NRF2 transcriptionally activated Lin28a in A172 cells. Bone marrow mesenchymal stromal cell-derived exosomes increased Lin28a expression in a NRF2-dependent manner. Bone marrow mesenchymal stromal cell-derived exosomal NRF2 improved OGD/R-induced A172 cell injury by inducing Lin28a production. Conclusion: Bone marrow mesenchymal stromal cell-derived exosomal NRF2 improved CIRI by transcriptionally activating Lin28a.

Up-regulation of lncRNA NEAT1 in cerebral ischemic stroke promotes activation of astrocytes by modulation of miR-488-3p/RAC1

We aim to research the molecular mechanism of lncRNA NEAT1 in the activation of astrocytes in a cerebral ischemia-reperfusion injury model. Mouse model of cerebral ischemia-reperfusion injury was constructed, and shNEAT1 was transfected. The infarct area, brain water content, and neurological deficiency were detected. Immunofluorescence detection and fluorescence in situ hybridization (FISH) assay were processed to detect glial fibrillary acidic protein (GFAP) expression. Astrocyte cells were cultured for oxygen-glucose deprivation/re-oxygenation (OGD)/re-oxygenation model construction. After treatment by shNEAT1, miR-488-3p mimic, miR-488-3p inhibitor, Q-PCR assay, western blot and ELISA were undertaken to detect the expressions of NEAT1, miR-488-3p, RAC1, inflammatory cytokines, RAC1 and GFAP. Dual luciferase reporter assay and RNA-binding protein immunoprecipitation (RIP) assay were used to verify the binding of NEAT1, miR-488-3p and RAC1. The expression of NEAT1 in brain tissue was significantly higher than that in Sham operation group. Knockdown of NEAT1 inhibited the brain damage caused by middle cerebral artery occlusion (MCAO) treatment, reduced the inflammatory response, and suppressed the activation of astrocytes. By constructing an in vitro OGD/R cell model, it was found that NEAT1 knockdown also inhibited the activation of astrocytes caused by OGD/R. Knockdown of NEAT1 caused the up-regulation of miR-488-3p and the down-regulation of RAC1. Knockdown of miR-488-3p or over-expression of RAC1 reversed the inhibitory effect of shNEAT1 on OGD/R-induced astrocyte activation. Over-expression of NEAT1 in cerebral ischemic stroke promotes activation of astrocytes by modulation miR-488-3p/RAC1, which is proved in vitro. Our study may provide a new idea for the diagnosis and treatment of MCAO.

Protective Effects of Nuciferine in Middle Cerebral Artery Occlusion Rats Based on Transcriptomics

Middle cerebral artery occlusion (MCAO), with the characteristics of high morbidity, high recurrence rate, high mortality, and disability rate, is a typical manifestation of ischemic stroke and has become a hot research topic in the clinical field. The protective effects of nuciferine on brain injury MCAO rats were investigated and its mechanisms of actions were revealed. The MCAO rats were established by the suture method. The pathological staining of the rat brain was processed and observed, the pharmacodynamics assay of nuciferine were studied, and the gene expression regulation by nuciferine was detected by transcriptome technology. The results showed that nuciferine significantly alleviated brain damage in MCAO rats, and the transcriptomic results suggested that nuciferine could exert therapeutic effects through the regulation of lipid metabolism, including arachidonic acid metabolism, sphingolipid metabolism, the PPAR signaling pathway and other related pathways. This finding provided new perspectives on the treatment of MCAO with nuciferine and facilitates the development of novel drugs for this disease.

In Vitro Differentiation of Bone Marrow Mesenchymal Stem Cells into Neuron-Like Cells by Cerebrospinal Fluid Improves Motor Function of Middle Cerebral Artery Occlusion Rats.

Bone marrow mesenchymal stem cells (BMSCs) represent a promising tool for stem cell-based therapies. However, the majority of BMSC transplants only allow for limited recovery of the lost functions. We previously found that human cerebrospinal fluid (hCSF) is more potent than growth factors in differentiating human BMSCs into neuron-like cells in vitro. In this study, we studied the effect of transplantation of rat BMSC-derived neuron-like cells (BMSC-Ns) induced by hCSF into rat brain with middle cerebral artery occlusion (MCAO). The survival and differentiation of the transplanted cells were determined using immunofluorescence staining of bromodeoxyuridine. The recovery of neurological function were observed by the modified neurological severity score (modified NSS) at 4, 15, and 32 days after cell transplantation, HE staining for determination of the infarct volume at day 32 after cell transplantation. Transplantation of BMSC-Ns or BMSCs significantly improved indexes of neurological function and reduced infarct size in rats previously subjected to MCAO compared with those in the control group. Remarkably, 32 days after transplantation, rats treated with BMSC-Ns presented a smaller infarct size, higher number of neuron-specific, enolase-positive, and BrdU-positive cells, and improved neurological function compared with BMSC group. Our results demonstrate that transplantation of hCSF-treated BMSC-Ns significantly improves neurological function and reduces infarct size in rats subjected to MCAO. This study may pave a new avenue for the treatment of MCAO.

The Safety and Efficacy of Continuous Transcranial Duplex Doppler Monitoring of Middle Cerebral Artery Occlusion in Acute Stroke Patients: Comparison of TCDD and Thrombolysis in MCA Recanalization

Assess the safety and efficacy of the continuous transcranial duplex Doppler (TCDD) monitoring of middle cerebral artery (MCA) (M1-2) occlusion in acute ischemic stroke (IS) patients and compare TCDD to intra-arterial thrombolysis (IAT) and intravenous thrombolysis (IVT). Forty consecutive acute IS patients were analyzed. Standard IVT was performed within 3 hours since stroke onset in 20 patients; between 3 and 6 hours continual 60 minute TCDD monitoring of occluded MCA using 2-MHz probe was performed in 10 patients and IAT in 10 patients. Neurological deficit was evaluated using the National Institutes of Health Stroke Scale and clinical outcome using modified Rankin Scale. The incidence of symptomatic intracerebral hemorrhage (sICH), clinical outcomes, and recanalization rates were compared among TCDD, IVT, and IAT. Analysis of variance, Kruskal-Wallis, and chi(2) tests were used for statistical evaluation. Incidence of sICH was 0% in TCDD group, 5% in IVT, and 20% in IAT (P= .198). Good 90-day clinical outcome (mRS 0-2) was achieved in 70% of TCDD patients (P= .570); recanalization after TCDD was found in 60% of patients (IVT 45%, IAT 70%) (P= .185). Continual TCDD monitoring might be safe and potentially beneficial in treatment of MCA occlusion.

Emergency embolectomy for treatment of acute middle cerebral artery occlusion

The authors evaluated the efficacy of emergency embolectomy in patients with acute middle cerebral artery occlusion (MCAO). A retrospective review of the charts for patients undergoing embolectomy at the authors' institution was performed. Between October 1997 and May 2004, 12 patients (mean age 70 years) with acute MCAO were treated using embolectomy. Local intraarterial fibrinolysis with urokinase was initially undertaken in eight of 12 patients. Sufficient recanalization was not achieved with fibrinolysis in any patient, and thus embolectomy was performed immediately thereafter. Recanalization by embolectomy was achieved in all patients (mean occlusion time 6 hours, 11 minutes). Each patient's condition was evaluated on discharge. Outcomes in the 12 patients according to the Glasgow Outcome Scale were good recovery in five, moderate disability in two, severe disability in three, vegetative state in one, and death due to a cardiac complication in one. None of the 12 patients had symptomatic hemorrhagic infarction. Emergency embolectomy can be performed in patients with MCAO with minimal morbidity and death. The procedure can be used to achieve good recovery even in patients in whom fibrinolysis is insufficient after acute MCAO and should be a part of the algorithm for the treatment of MCAO.

Expression of insulin-like growth factor 1 and receptor in ischemic rats treated with human marrow stromal cells

Human bone marrow stromal cells (hMSCs) enhance neurological recovery after stroke in rodents, possibly via induction of growth factors. We therefore elected to test the effects of hMSC treatment on insulin-like growth factor 1 (IGF-1), which plays an important role in growth, development, neuroprotection and repair in the adult. Rats ( n=57) were subjected to permanent middle cerebral artery occlusion (MCAo) and injected intravenously with 3×10 6 hMSCs or phosphate-buffered saline (PBS) at 1 day after MCAo. Functional outcome was measured after MCAo using a modified Neurological Severity Score (mNSS). Gene expression of IGF-1 and IGF-1 receptor (IGF-1R) in the ischemic brain tissue were measured at 2 and 7 days after MCAo using reverse transcription–polymerase chain reaction (RT-PCR). Immunohistochemistry was performed to measure the expression of bromodeoxyuridine (BrdU), doublecortin (DCX), IGF-1 and IGF-1R at 7, 14 and 30 days after MCAo. Treatment of MCAo with hMSCs significantly improved functional recovery from 14 to 30 days. MAB1281-labeled hMSCs entered the ischemic brain and increased time-dependently. hMSC treatment significantly increased IGF-1 mRNA and BrdU +, DCX +, IGF-1 + and IGF-1R + cells compared to PBS-treated rats ( p<0.05). The percentage of BrdU + or DCX + cells colocalized with IGF-1 increased in the hMSC-treated rats compared to the PBS-treated rats ( p<0.05). IGF-1 and IGF-1R may contribute to improved functional recovery and increased neurogenesis after treatment of stroke with hMSCs.