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Liver transplantation is a vital treatment for end-stage liver disease. However, the shortage of donor livers has limited the development of liver transplantation. How to expand the source of donor livers has become a challenge in the academic community. In recent years, the proportion of donors with non-alcoholic fatty liver disease (NAFLD) has been increased. Rational use of steatotic donor livers is a feasible approach to expand the donor pool. Cold ischemia injury during donor liver preservation before liver transplantation increases the risk of postoperative organ dysfunction. Therefore, it is of significance to unravel the mechanism and intervention measures of cold ischemia injury of steatotic donor livers. Cold ischemia injury of steatotic donor livers is characterized as the damage of mitochondria, lysosomes and endoplasmic reticulum at the organelle level, and up-regulated expression of adenosine monphosphate activated protein kinase (AMPK), aldehyde dehydrogenase 2 (ALDH2) and heme oxygenase (HO)-1 at the protein level. In this article, the research progresses on cold ischemia injury of steatotic donor livers and relevant intervention measures were reviewed.
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Objective To observe the protective effect of Shenmajing formula on brain tissue of mice with cerebral ischemic injury and explore the possible mechanism. Methods Thirty SPF-grade C57 BL/6 male mice were randomly divided into model control group, Shenmajing group and nimodipine group, and the animal models of cerebral ischemic injury in mice were prepared by electrocoagulation. The protein expression level in endothelial progenitor cells were detected by Western blot. Results Compared with the model control group, the infarct volume of mice in the Shenmajing group was significantly reduced, and the migration, adhesion and tubule formation ability of endothelial progenitor cells were significantly improved, and the expression level of BDNF protein in endothelial progenitor cells was significantly increased. Conclusion The protective effect of Shenmajing granules on brain tissue of mice with cerebral ischemic injury could be closely related to the regulation of BDNF expression in endothelial progenitor cells and improvement of endothelial progenitor cell function of bone marrow origin.
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Abstract Objective: Given the high proliferative activity of germinal matrix and its direct correlation with hypoxemia, it is necessary to investigate the possible molecular regulation pathways, to understand the existing clinical relationship between the hypoxic-ischemic insult and the biomarkers NF-kB, AKT-3, Parkin, TRK-C and VEGFR-1. Methods: A hundred and eighteen germinal matrix samples of the central nervous system of patients who died in the first 28 days of life were submitted to histological and immunohistochemistry analysis to identify the tissue immunoexpression of those biomarkers related to asphyxia, prematurity, and death events within 24h. Results: A significantly increased tissue immunoexpression of NF-kB, AKT-3 and Parkin was observed in the germinal matrix of preterm infants. In addition, significantly decreased tissue immunoexpression of VEGFR-1 and NF-kB was observed in patients who experienced asphyxia followed by death within 24 hours. Conclusions: The results suggest a direct involvement between the hypoxic-ischemic insult and NF-kB and VEGFR-1 markers since a decreased immunoexpression of these biomarkers was observed in asphyxiated patients. Furthermore, it is suggested that there was not enough time for VEGFR-1 to be transcribed, translated and expressed on the surface of the plasma membrane. This temporality can be observed in the relationship between NF-kB expression and the survival time of individuals who died within 24 hours, suggesting that this factor is essential for the production of VEGFR-1 and, therefore, to carry out the necessary remodeling effect to neovascularize the affected region.
RESUMO Objetivo: Dada a alta atividade proliferativa da matriz germinativa e sua correlação direta com a hipoxemia, é necessário investigar as possíveis vias de regulação molecular para entender a relação clínica existente entre o insulto hipóxico-isquêmico e os biomarcadores NF-kB, AKT -3, Parkina, TRK-C e VEGFR-1. Métodos: Cento e dezoito amostras de matriz germinativa do sistema nervoso central de pacientes que faleceram nos primeiros 28 dias de vida foram submetidas a análise histológica e imuno-histoquímica para identificar a imunoexpressão tecidual desses biomarcadores relacionados a eventos de asfixia, prematuridade e óbito em 24 horas. Resultados: Observou-se uma imunoexpressão tecidual significativamente aumentada de NF-kB, AKT-3 e Parkin na matriz germinativa de prematuros. Além disso, constatou-se uma imunoexpressão tecidual significativamente diminuída de VEGFR-1 e de NF-kB em pacientes que apresentaram asfixia seguida de morte em 24 horas. Conclusões: Os resultados sugerem o envolvimento direto entre o insulto hipóxico-isquêmico e os marcadores NF-kB e VEGFR-1, visto que se observou uma imunoexpressão diminuída destes biomarcadores nos pacientes asfixiados. Além disso, sugere-se que não houve tempo suficiente para que o VEGFR-1 fosse transcrito, traduzido e expresso na superfície da membrana plasmática. Essa temporalidade pode ser observada na relação entre a expressão de NF-kB e o tempo de vida dos indivíduos que morreram em 24 horas, o que sugere que esse fator é essencial para a produção do VEGFR-1 e, portanto, para realizar o efeito remodelador necessário para neovascularizar a região afetada.
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Resumo Fundamento: Embora a elevação não isquêmica da troponina seja frequentemente observada em pacientes admitidos no pronto-socorro (PS), não há consenso quanto ao seu manejo. Objetivos: Este estudo teve como objetivo caracterizar os pacientes admitidos no PS com elevação da troponina não-isquêmica e identificar potenciais preditores de mortalidade nessa população. Métodos: Este estudo observacional retrospectivo incluiu pacientes do PS com resultado positivo no teste da troponina entre junho e julho de 2015. Pacientes com diagnóstico clínico de síndrome coronariana aguda (SCA) foram excluídos. Os dados demográficos dos pacientes e as variáveis clínicas e laboratoriais foram extraídos dos prontuários médicos. Os dados do seguimento foram obtidos por 16 meses ou até a ocorrência de morte. O nível de significância estatística foi de 5%. Resultados: A elevação da troponina sem SCA foi encontrada em 153 pacientes no PS. A mediana (IIQ) de idade dos pacientes foi de 78 (19) anos, 80 (52,3%) eram do sexo feminino e 59 (38,6%) morreram durante o seguimento. A mediana do período de seguimento (IIQ) foi de 477 (316) dias. Os sobreviventes eram significativamente mais jovens 76 (24) vs. 84 (13) anos; p=0,004) e apresentaram uma maior proporção de elevação da troponina isolada (sem elevação da creatina quinase ou mioglobina) em duas avaliações consecutivas: 48 (53,9%) vs. 8 (17,4%), p<0,001. Os sobreviventes também apresentaram menor taxa de tratamento antiplaquetário e internação no mesmo dia. Na regressão logística multivariada com ajuste para variáveis significativas na análise univariada, a elevação isolada da troponina em duas avaliações consecutivas mostrou hazard ratio = 0,43 (IC95% 0,17-0,96, p=0,039); hospitalização, tratamento antiplaquetário anterior e idade permaneceram independentemente associados à mortalidade. Conclusões: A elevação isolada da troponina em duas medidas consecutivas foi um forte preditor de sobrevida em pacientes no PS com elevação da troponina, mas sem SCA.
Abstract Background: Although non-ischemic troponin elevation is frequently seen in patients admitted to the emergency department (ED), consensus regarding its management is lacking. Objectives: This study aimed to characterize patients admitted to the ED with non-ischemic troponin elevation and to identify potential mortality predictors in this population. Methods: This retrospective observational study included ED patients with a positive troponin test result between June and July of 2015. Patients with a clinical diagnosis of acute coronary syndrome (ACS) were excluded. Data on patient demographics and clinical and laboratory variables were extracted from medical records. Follow-up data were obtained for 16 months or until death occurred. The statistical significance level was 5%. Results: Troponin elevation without ACS was found in 153 ED patients. The median (IQR) patient age was 78 (19) years, 80 (52.3%) were female and 59(38.6%) died during follow-up. The median (IQR) follow-up period was 477(316) days. Survivors were significantly younger 76 (24) vs. 84 (13) years; p=0.004) and featured a higher proportion of isolated troponin elevation (without creatine kinase or myoglobin elevation) in two consecutive evaluations: 48 (53.9%) vs. 8 (17.4%), p<0.001. Survivors also presented a lower rate of antiplatelet treatment and same-day hospitalization. In the multivariate logistic regression with adjustment for significant variables in the univariate analysis, isolated troponin elevation in two consecutive evaluations showed a hazard ratio= 0.43 (95%CI 0.17-0.96, p=0.039); hospitalization, previous antiplatelet treatment and age remained independently associated with mortality. Conclusions: Isolated troponin elevation in two consecutive measurements was a strong predictor of survival in ED patients with troponin elevation but without ACS.
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Humans , Male , Female , Aged , Troponin I , Acute Coronary Syndrome/diagnosis , Prognosis , Biomarkers , Emergency Service, Hospital , HospitalizationABSTRACT
Background: Seizure is a pediatric emergency. Accurate determination of the etiology of seizures is very important to start an effective treatment. The study aims to determine the spectrum of Imaging abnormalities by Magnetic Imaging Resonance (MRI) in children who presented with seizures. Methods: It is a hospital-based prospective observational study which was carried out in Government Medical College and Rajindra Hospital, Patiala. This study included 50 pediatric patients in the age group between 0 months to 18 years who were referred to the Department of Radiodiagnosis for brain MRI between October 2017 to September 2019. Results: Neuroimaging abnormality was found in 19 (38%) cases. 31 (62%) patients had no abnormal finding. The most common imaging findings were inflammatory granuloma in 5 (10%) patients. Other findings were- Hypoxic-ischemic injury (HII) in 4 (8%), Mesial temporal sclerosis in 2(4%), cerebral atrophy in 1(2%), Hemorrhage in 1(2%), Tuberous sclerosis in 1(2%), Focal cortical dysplasia in 1(2%), Lissencephaly in 1 (2%), Joubert syndrome in 1(2%), and Arachnoid cyst in 1 (2%) patients. Conclusion: The MRI was able to identify brain lesions in 38% of pediatric patients who presented with seizures. The study revealed inflammatory granuloma as the commonest cause of seizures in children, followed by Hypoxic-Ischemic Injury. Early recognition of potentially treatable diseases helps in timely treatment and arrest of disease progression. It is recommended to use MRI as a primary investigation during the evaluation and management of pediatric seizures.
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OBJECTIVE: To investigate the effect of electroacupuncture (EA) on the expression of interleukin-8 (IL-8), interleukin-10 (IL-10), tyrosine hydroxylase (TH), β3-adrenergic receptor (β3AR), and endothelial nitric oxide synthase (eNOS) in myocardial tissue in ischemic myocardial injury rats, so as to reveal its underlying mechanisms in myocardial protection via anti-inflammation and sympathetic nerve remodeling. METHODS: A total of 48 male Sprague-Dawley rats were randomly divided into sham-operation (sham, n＝9), sham ＋EA (n＝9), model (n＝15) and EA (n＝15) groups. The myocardial ischemia (MI) model was established by ligation of the left anterior descending branch of the left coronary artery. EA (2 Hz/15 Hz，1.5－2 mA) was applied to bilateral "Neiguan" (PC6) for 30 min, once daily for 4 days. The myocardial infarct size was detected by 2, 3, 5 triphenyltetrazolium chloride (TTC) staining, myocardial histopathological changes and inflammatory infiltration were assessed by H．E. staining, and the expression of IL-8, IL-10, TH, β3AR, and eNOS in the myocardium was determined by using Western blot. RESULTS: Compared with the sham group, a marked myocardial infarction was found in the left ventricle tissue, accompanied with disordered arrangement of myocardial fibers and higher degree of inflammatory cell infiltration, and increased expression of IL-8, TH, β3AR and eNOS in the myocardium in the model group (P0.05). After EA intervention and in comparison with the model group, the myocardial infarct size was significantly reduced (P<0.01), the severity of inflammatory cell infiltration and disordered arrangement of myocardial fibers were relieved, and the expression of IL-10 and eNOS proteins were significantly up-regulated (P<0.05), and the markedly up-regulated expression of IL-8, TH, and β3AR were significantly suppressed in the EA group (P<0.01)．. CONCLUSION: EA intervention can reduce the myocardial infarct size (protective effect) in MI rats possibly by reducing inflammatory reaction and sympathetic nerve remodeling.
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Objective:To investigate the protective effect of Yiqi Huoxue recipe on rats with cerebral ischemia injury by using oxidative stress injury as an entry point. Method:SD rats were randomly divided into model group, sham operation group, nimodipine group (20 mg·kg-1), Yiqi Huoxue recipe high, medium and low dose group (2.916,1.458,0.729 g·kg-1). After 14 days of stomach, acute cerebral ischemic injury model was established by ligation of bilateral common carotid arteries. Ultrasound of synapse was observed by transmission electron microscopy. Total superoxide dismutase (T-SOD) and dialdehyde (MDA), lactate dehydrogenase (LDH), glutathione peroxidase (GSH-Px), horizontal adenine nucleoside triphosphate (ATP) levels were detected by biochemical method. Western blot and Real-time PCR was used to determine the expression of heme oxygenase-1(HO-1) and nuclear factor E2-related factor 2 (Nrf2) protein and mRNA in the ischemic cortex of rats. Result:Transmission electron microscopy showed that Yiqi Huoxue recipe had a significant improvement on the degree of cerebral ischemic injury. Compared with sham operation group, MDA levels in the brain homogenate of model group increased significantly, T-SOD and GSH-Px levels were significantly decreased (P+-K+-ATP ase, Ca2+-Mg2+-ATP ase and ATP was significantly decreased (PPPPP+-K+-ATP ase, Ca2+-Mg2+-ATP ase and total ATP activity(PPPPPPConclusion:Yiqi Huoxue recipe may protect against cerebral ischemic injury by inhibiting oxidative stress through Nrf2/HO-1 signaling pathway.
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Objective To investigate the effects of bone marrow mesenchymal stem cells (BMSCs) transplantation on the expression of nerve growth factor (NGF) and Caspase-3 in rat hippocampus after cardiac arrest (CA). Methods Sprague-Dawley (SD) rats were randomly divided into 3 groups: sham group (n=6), CA group (n=6), and BMSCs group (n=6). CPR was performed on the groups after the induction of asphyxial cardiac arrest. Animals in the BMSCs group or the CA group were respectively injected with a dose of 1×106 BMSCs in 0.5 mL phosphate buffer solution (PBS) or 0.5 mL PBS alone via the vena caudalis 1 h after successful resuscitation. The neurological status after restoration of spontaneous circulation (ROSC) were assessed by modified neurological severity scores (mNSS); serum levels of S100B were assayed, and the expression of NGF and Caspase-3 in hippocampus was detected by immunohistochemistry. Results Compared with the CA group, mNSS and S100B levels were lower in the BMSCs group on the 7th day after ROSC [(0.9±0.3) vs (4.5±0.6), (90.12±4.62) pg/mL vs (182.30±2.58) pg/mL, both P<0.05] with higher expression of NGF and lower expression of Caspase-3 [(11.391±1.297) vs (7.744±1.334), (6.256±1.036) vs (8.506±1.742), both P< 0.05]. Conclusions BMSCs transplantation might improve rat's neurological functions after cardiac arrest, which may be related to up-regulation of NGF expression and down-regulation of Caspase-3 expression.
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Objective To study the metabolic process of ginkgolides in rats with cerebral ischemic injury based on pharmacokinetic- pharmacodynamics (PK-PD) binding model. Methods The middle cerebral artery occlusion (MCAO) model was established by the suture method. After reperfusion, rats were randomly assigned to nasal administration, ig administration, and iv administration group.The orbital blood was taken at different time points of 0.25, 0.33, 0.5, 0.75, 1.0, 1.25, 1.5, 2.0, 4.0, 6.0, 9.0, and 12.0 h after the administration of the ginkgolides solution. The drug-time curve of ginkgolide B in plasma were drawn according to the concentration measured by LC-MS. The time-effect curve of superoxide dismutase (SOD) and malondialdehyde (MDA) were drawn based on the value measured by the kit. The pharmacokinetic parameters were calculated by DAS 2.0 software to fit the PK-PD binding model. Results The t1/2 of ginkgolide B of the rats in the administration group was smaller than that in the MCAO model group. The area under the curve (AUC) of nasal administration was significantly higher than intragastric administration and intravenous administration. Conclusion Ginkgolide B has a good protective and mitigating effect on ischemic stroke. The pharmacokinetics of nasal administration is better than iv and ig administration, which can provide reference for the development of nasal administration of ginkgolide B.
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The in-depth study on the law of drug pair compatibility and its mechanism has important scientific connotation for the clinical prescriptions and the improvement of curative effect. In recent years, many prevention and treatment of cerebral ischemic injury found that the rational use of Chinese materia medica pairs by optimizing the combination of Chinese medicine compound played a multi-target, multi-level role in the ischemic brain tissue of the neurovascular units, multi-channel regulation of the relevant signal pathways, which can significantly reduce the damage of ischemic penumbra brain tissue, relieve the inflammatory cascade and reperfusion injury caused by cerebral ischemia injury, promptly restore the blood flow in the brain and effectively protect the neurons, achieve “re-flow” and “brain protect” similar effects, and further promote the repair of nerve function.
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AIM To observe the oxidant stress and opoptotic effects of anisodine hydromide (AH) on chronic cerebral hypoperfusion (CCH) rats.METHODS In vivo CCH models were established in adult male SpragueDawley rats by permanent ligation of bilateral common carotid arteries [two-vessel occlusion (2-VO)] surgery.Rats were randomly divided into six groups,sham group,model group,positive group of n-butylphthalide and sodium chloride injection,and AH groups (1.2 mg/kg high-dose group,0.6 mg/kg medium-dose group,and 0.3 mg/kg low-dose group).Antioxidant indices including the activity of SOD,CAT,LDH and iNOS and the content of GSH and NO were measured.In the in vitro trial,PC12 cells were divided into control group,model group,positive group of n-butylphthalide,and AH groups (100 μmol/L high-dose group,50 μmol/L mediumdose group,and 25 μmol/L low-dose group),and the hypoxic models were established by treating PC12 cells with CoCl2.The cells had their release of NO and LDH detected,their cellular apoptosis determined by Hochest 33342 fluorescence staining,and the expression of P53 protein identified by IF (immunofluorescence) and Western blotting method.RESULTS The in vivo trial revealed AH's enhancement in serum SOD activity and inhibition in serum iNOS activityof the CCH rats,and its power in the cerebral GSH and LDH release reduction.The in vitro trial showed the resultant lower LDH and NO release,decreased number of neuro-apoptosis,and inhibited P53 pro tein expression after AH intervention.CONCLUSION The antioxidant and antiapoptotic effects of AH on CCH rats may be associated with down regulation of P53 protein.
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Fibroblast growth factor 21 (FGF21) is regarded as an important metabolic regulator playing a therapeutic role in diabetes and its complications.Recently, growing evidence showed that heart is a key target as well as a source of FGF21 which is involved in heart development and also induces beneficial effects in cardiovascular diseases.Strong evidences showed that the development of cardiovascular diseases including atherosclerosis, atherosclerosis, coronary heart disease, myocardial ischemia, cardiac hypertrophy, and diabetic cardiomyopathy is associated with increased serum FGF21 levels which was regarded as a compensatory response to induced cardiac protection.This review tries to illuminate the underlying relationship between FGF21 and cardiovascular diseases and the possible mechanisms.
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AIM To investigate the anti-neuroinflammation effects of 4-hydroxybenzyl aldehyde (4-HBAL) from Gastrodia elata Blume on acute cerebral ischemic injury in rats and its nechanism of action.METHODS The rat model of acute cerebral ischemic injury was induced by injecting arachidonic acid via intracarotid artery.Brain tissue samples were taken from the animals 3 h after the model of acute cerebral ischemic injury.Tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β) were detected in brain tissue to evaluate the effects of 4-HBAL in vivo.Lipopolysaccharid (LPS)-induced activation of BV-2 microglia cells model was used to explore the anti-neuroinflammation mechanism of 4-HBAL.RESULTS The experimental results showed that 4-HBAL had a significant protective effect on acute cerebral ischemic injury.It could significandy decrease the contents of tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β),and obviously inhibit the production of nitric oxide (NO),prostaglandin E2 (PGE2) and TNF-α in LPS-stimulated BV-2 cell,and increase the production of interleukin-10 (IL-10) and transforming growth factors-β (TGF-β) in BV-2 cell.CONCLUSION The mechanism of 4-HBAL may be related to the suppression of the excessive activation of microglia after cerebral ischemia and the promotion of the transformation of microglia into anti-inflammatory phenotype.
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Objective To estabhsh an ischemic model of intratemporal facial nerve (IFN) via the mastoid process approach.Methods From February,2015 to December,2015,45 SD rats were divided randomly into an operation group (n=35) and a shame group (n=10) in random,the right side facial nerve was used for the operation and the left side served as the control in both groups.Establish the IFN ischemia model by interrupting the petrosal artery through the mastoid process approach.Facial nerve function were evaluated at the 12h and everyday postoperatively for 28 days.The degree of IFN swelling were studied by taking paraffin sections of the decalcified temporal bone containing the IFN instantly and at the 1st,3rd,7th,14th and 21st days postoperatively.Then calculated the ratio of the cross-sectional area of the IFN and the facial canal (FN/FC).The data of behavior assessment and FC/FN were analyzed using ANOVA.Twenty-eight days after the insult,took continuous sections of brainstem containing facial nucleus,then counted the number of the facial neurons.At last,analysed the results of both operation and control sides in each group by using the student-t test.Results Facial nerve paralysis developed at 12 hour after surgery,then continued deteriorated till the 7th day.By the 28th day postoperatively,all rats in surgery group recovered and data showed no significance statistically when compared with the shame group (P＜0.05).From the value of FN/FC in different groups,the nerve were found swelling in the facial canal was increasing from the 1st postoperatively and reach the peak value at the 7th day after surgery.By the 21st day,the FN/FC come to steady but remain significant statistically when compared with the contralateral side(P＜0.05).In the operation group,facial neurons of injury side exhibited significantly loss [(41.5±3.8)％] when compared with the shame group [(98.1±4.0)％](P＜0.05).Conclusion Rats with petrosal artery interrupted exhibited significant deficits.This approach involved less tissue injury,studies on the mechanisms and therapy could become more reliable using this approach.
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This study aimed to explore the neuroprotection and mechanism of isoflurane on rats with spinal cord ischemic injury. Total 40 adult male Sprague-Dawley rats were divided into the four groups (n=10). Group A was sham-operation group; group B was ischemia group; group C was isoflurane preconditioning group; group D was isoflurane preconditioning followed by ischemia treatment group. Then the expressions of TWIK-related K⁺ channel 1 (TREK1) in the four groups were detected by immunofluorescent assay, real time-polymerase chain reactions (RT-PCR) and western blot. The primary neurons of rats were isolated and cultured under normal and hypoxic conditions. Besides, the neurons under two conditions were transfected with green fluorescent protein (GFP)-TREK1 and lentivirual to overexpress and silence TREK1. Additionally, the neurons were treated with isoflurane or not. Then caspase-3 activity and cell cycle of neurons under normal and hypoxic conditions were detected. Furthermore, nicotinamide adenine dinucleotide hydrate (NADH) was detected using NAD+/NADH quantification colorimetric kit. Results showed that the mRNA and protein expressions of TREK1 increased significantly in group C and D. In neurons, when TREK1 silenced, isoflurane treatment improved the caspase-3 activity. In hypoxic condition, the caspase-3 activity and sub-G1 cell percentage significantly increased, however, when TREK1 overexpressed the caspase-3 activity and sub-G1 cell percentage decreased significantly. Furthermore, both isoflurane treatment and overexpression of TREK1 significantly decreased NADH. In conclusion, isoflurane-induced neuroprotection in spinal cord ischemic injury may be associated with the up-regulation of TREK1.
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Adult , Animals , Humans , Male , Rats , Blotting, Western , Caspase 3 , Cell Cycle , Ischemia , Isoflurane , Models, Animal , NAD , Neurons , Neuroprotection , Rats, Sprague-Dawley , RNA, Messenger , Spinal Cord , Up-RegulationABSTRACT
AIM:To explore the role of microRNA-181b (miR-181b) in ischemic injury and autophagy pro-tein 5 (Atg5) levels of mice .METHODS:Oxygen-glucose depletion (OGD) model in N2A cells to mimic ischemic in-jury in vitro was established .A middle cerebral artery occlusion ( MCAO) model to mimic ischemic injury in vivo was also induced in mice.The N2A cell apoptosis after OGD was assessed by in situ cell death detection kit.The Atg5 and caspase-9 expressions were determined by Western blotting .Luciferase reporter assay was performed to identify the direct binding of miR-181b with 3’-UTR of Atg5 mRNA.RESULTS:The alteration of miR-181b expression level by transfection with pre-miR-181b or anti-miR-181b significantly affected N2A cell apoptosis (P<0.05).Accordingly, the changes of miR-181b levels significantly altered the protein level of Atg 5 ( P<0.05 ) .Co-transfection of the luciferase reporters with pre-miR-181b or anti-miR-181b resulted in the inhibition or enhancement of the luciferase activities of luciferase expressing plasmid containing 3’-UTR of Atg5 mRNA (P<0.05).In addition, the miR-181b antagonist significantly reduced the cleaved caspase-9 levels in cerebral ischemic cortex of the mice after MCAO ( P<0.05 ) .CONCLUSION: Down-regulation of miR-181b plays an important role in ischemic injury of mice through regulating Atg 5 protein level.
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Objective To explore the role of miR-184 in Oxygen-Glucose-Deprivation (OGD) induced SK-N-SH cell ischemic injury and its regulation on AKT2 level. Method We used a combination of oxygen and glucose deprivation to imitate ischemic conditions in vivo. MiR-184 mimic and inhibitor were transfected into SK-N-SH cell to alter miR-184 levels. The expression of miR-184 and AKT2 were determined by using Real-time PCR. The extent of SK-N-SH cell survival rate was assessed by thiazolyl blue tetrazolium bromide (MTT) assay. Result Here, we observed that miR-184 was significantly inhibited in SK-N-SH cell after OGD (P<0.05). The changes of miR-184 level altered the expression of AKT2 mRNA. In addition, alteration of miR-184expressionsignificantly affected cell survival rate after OGD. Conclusion miR-184 plays an important role in ischemic injury through negatively regulating AKT2 level, which may provide a potential therapeutic target for ischemic stroke in miRNA levels.
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Objective To investigate the preventive effects of Buyang Huanwu Decoction (BYHWD) and its recipe composition (BYJJF) in focal ischemic brain injury condition in vivo/in vitro. Methods In vivo studies, SD rats were divided into sham-operation group, MCAO group, BYHWD group and BYJJF group based on rat weight, 10 rats in each group. The body weight, infarct area and brain water contents were determined. In vitro studies, H2O2 was used to damage PC12 cells, and the vitro oxidative stress cell model was established. PC12 cells were divided into normal group, blank control group, BYHWD and BYJJF groups with different concentrations (0.1, 0.2, 0.5, 1.0, 2.0, 3.5 mg/mL). MTT method was employed to determine the protective effects of BYHWD and BYJJF on model cells.Results Vivo studies showed that after 7 days of treatment with BYHWD and BYJJF, those determinated quotas were all significantly improved compared with MCAO model rats (P0.05).Vtiro studies showed that the protective effects of BYHWD and BYJJF took place 2 hours later, and it was obvious in oxidative stress injury caused by H2O2, with statistical differences with model group (P0.05).Conclusion The research confirmed that BYJJF plays a significant role in improving the cerebral ischemia injury, which is the same performance as BYHWD, and BYJJF can save TCM resources under the precondition of TCM efficacy.
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Aim To observe the neuroprotective effect of sinomenine on hippocampal neurons from injury in-duced by oxygen glucose deprivation ( OGD ) and its underlying mechanism. Methods Hippocampal neu-rons were exposed to OGD for 4 h followed by 24 h re-oxygenation ( OGD-R) . Then cell viability was detec-ted by MTT. LDH release was detected by LDH kit. Cell apoptosis was detected by Hoechst stain. The ex-pression of Bax, Bcl-2 and caspase-3 were detected by Western blot. [ Ca2+] i of hippocampal neurons was detected by calcium imaging. Acid-sensing ion chan-nels ( ASICs ) current was detected by patch clamp technique. Results SN increased cell viability and reduced LDH release. SN also inhibited neuron apop-tosis and increased ratio of Bcl-2/Bax and reduced the expression of caspase-3 . OGD-induced increase of [ Ca2+] i was inhibited by SN. Furthermore, SN inhib-ited ASIC1 a current and also inhibited OGD induced increase of ASICs current in hippocampal neurons. Conclusion SN protects hippocampal neurons against OGD-R-induced injury. The inhibitory effect of SN on ASIC1 a and calcium overload was involved in the pro-tective effect of SN.
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Neural stem cells (NSCs) have been suggested as a groundbreaking solution for stroke patients because they have the potential for self-renewal and differentiation into neurons. The differentiation of NSCs into neurons is integral for increasing the therapeutic efficiency of NSCs during inflammation. Apoptosis signal-regulating kinase 1 (ASK1) is preferentially activated by oxidative stress and inflammation, which is the fundamental pathology of brain damage in stroke. ASK1 may be involved in the early inflammation response after stroke and may be related to the differentiation of NSCs because of the relationship between ASK1 and the p38 mitogen-activated protein kinase pathway. Therefore, we investigated whether ASK1 is linked to the differentiation of NSCs under the context of inflammation. On the basis of the results of a microarray analysis, we performed the following experiments: western blot analysis to confirm ASK1, DCX, MAP2, phospho-p38 expression; fluorescence-activated cell sorting assay to estimate cell death; and immunocytochemistry to visualize and confirm the differentiation of cells in brain tissue. Neurosphere size and cell survival were highly maintained in ASK1-suppressed, lipopolysaccharide (LPS)-treated brains compared with only LPS-treated brains. The number of positive cells for MAP2, a neuronal marker, was lower in the ASK1-suppressed group than in the control group. According to our microarray data, phospho-p38 expression was inversely linked to ASK1 suppression, and our immunohistochemistry data showed that slight upregulation of ASK1 by LPS promoted the differentiation of endogenous, neuronal stem cells into neurons, but highly increased ASK1 levels after cerebral ischemic damage led to high levels of cell death. We conclude that ASK1 is regulated in response to the early inflammation phase and regulates the differentiation of NSCs after inflammatory-inducing events, such as ischemic stroke.