Knockdown of miR-429 Attenuates Aβ-Induced Neuronal Damage by Targeting SOX2 and BCL2 in Mouse Cortical Neurons

Abstract Co-treatment of neuroprotective reagents may improve the therapeutic efficacy of hypothermia in protecting neurons during ischemic stroke. This study aimed to find promising drugs that enhance the neuroprotective effect of mild hypothermia (MH). 26 candidate drugs were selected based on different targets. Primary cultured cortical neurons were exposed to oxygen-glucose deprivation and reoxygenation (OGD/R) to induce neuronal damage, followed by either single treatment (a drug or MH) or a combination of a drug and MH. Results showed that, compared with single treatment, combination of MH with brain derived neurotrophic factor, glibenclamide, dizocilpine, human urinary kallidinogenase or neuroglobin displayed higher proportion of neuronal cell viability. The latter three drugs also caused less apoptosis rate in combined treatment. Furthermore, co-treatment of those three drugs and MH decreased the level of reactive oxygen species (ROS) and intracellular calcium accumulation, as well as stabilized mitochondrial membrane potential (MMP), indicating the combined neuroprotective effects are probably via inhibiting mitochondrial apoptosis pathway. Taken together, the study suggests that combined treatment with hypothermia and certain neuroprotective reagents provide a better protection against OGD/R-induced neuronal injury.

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MicroRNA-223 protects neurons from degeneration in Experimental Autoimmune Encephalomyelitis

10.1101/430777 ◽

2018 ◽

Author(s):

Barbara Morquette ◽

Camille A. Juźwik ◽

Sienna S. Drake ◽

Marc Charabati ◽

Yang Zhang ◽

...

Keyword(s):

Experimental Autoimmune Encephalomyelitis ◽

Cortical Neurons ◽

Neuronal Degeneration ◽

Neuronal Damage ◽

Messenger Rnas ◽

Autoimmune Encephalomyelitis ◽

Cns Inflammation ◽

Peripheral Blood Mononuclear ◽

The Brain ◽

Experimental Autoimmune

AbstractMultiple sclerosis (MS) is an autoimmune disease characterized by demyelination and neurodegeneration in the brain, spinal cord and optic nerve. Neuronal degeneration and death underlie progressive forms of MS and cognitive dysfunction. Neuronal damage is triggered by numerous harmful factors in the brain that engage diverse signalling cascades in neurons thus therapeutic approaches to protect neurons will need to focus on agents that can target broad biological processes. To target the broad spectrum of signaling events that mediate neurodegeneration in MS we have focused on non-coding small microRNAs (miRNAs). microRNAs are epigenetic regulators of protein expression, targeting messenger RNAs (mRNAs) and inhibiting their translation. Dysregulation of miRNAs has been described in many neurodegenerative diseases including MS. In this study we identified two miRNAs, miR-223-3p and miR-27a-3p, that were upregulated in neurons in the experimental autoimmune encephalomyelitis (EAE) mouse model of CNS inflammation and in active MS lesions. Overexpression of miR-27a-3p or miR-223-3p protected dissociated cortical neurons from degeneration in response to peripheral blood mononuclear cell conditioned media (PBMC-CM). Introduction of miR-223-3p in vivo in mouse retinal ganglion cells (RGCs) protected RGC axons from degeneration in the EAE model. By in silico analysis we found that mRNAs in the glutamate receptor (GluR) pathway are enriched in miR-27a-3p and miR-223-3p targets. Antagonism of the GluR pathway protected neurons from PBMC-CM-dependent degeneration. Our results suggest that miR-223-3p and miR-27a-3p are upregulated in response to inflammation to mediate a compensatory neuroprotective gene expression program that desensitizes neurons to glutamate by downregulating mRNAs involved in GluR signalling.

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Protective effect of Toki-shakuyaku-san on amyloidβ25-35-induced neuronal damage in cultured rat cortical neurons

Phytotherapy Research ◽

10.1002/ptr.1671 ◽

2005 ◽

Vol 19 (5) ◽

pp. 450-453 ◽

Cited By ~ 23

Author(s):

Nobuaki Egashira ◽

Katsunori Iwasaki ◽

Yuki Akiyoshi ◽

Yuki Takagaki ◽

Izzettin Hatip-Al-Khatib ◽

...

Keyword(s):

Protective Effect ◽

Cortical Neurons ◽

Neuronal Damage ◽

Rat Cortical Neurons

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Intravenous antagomiR-494 lessens brain-infiltrating neutrophils by increasing HDAC2-mediated repression of multiple MMPs in experimental stroke

10.21203/rs.2.17389/v1 ◽

2019 ◽

Author(s):

Fangfang Li ◽

Haiping Zhao ◽

Guangwen Li ◽

Sijia Zhang ◽

Rongliang Wang ◽

...

Keyword(s):

Cortical Neurons ◽

Neuronal Damage ◽

Neutrophil Migration ◽

Glucose Deprivation ◽

Neutrophil Infiltration ◽

Predictive Biomarker ◽

Experimental Stroke ◽

Mmp Genes

Abstract Background: Neutrophil infiltration and phenotypic transformation are believed to contribute to neuronal damage and clinical outcome in ischemic stroke. Emerging evidence suggests that HDAC2 is an epigenetic regulator of inflammatory cells. Here, we investigated whether miR-494 affects HDAC2-mediated neutrophil infiltration and phenotypic shift. Methods: The miR-494 levels in neutrophils from AIS patients were detected by real-time PCR. C57BL/6J mice were subjected to transient middle cerebral artery occlusion, and the N1/N2 neutrophil shift was examined. Cortical neurons were subjected to oxygen-glucose deprivation and stimulated with supernatant from differently treated neutrophils or were cocultured with neutrophils; neuronal injury was detected, and ChIP-Seq was performed to clarify which genes are the binding targets of HDAC2. Finally, a transwell assay was conducted to examine neutrophil migration. Results: Compared to the control subjects, AIS patients had increased neutrophil expression of miR-494, and in AIS patients, elevated miR-494 expression in neutrophils was a predictor of worse neurological outcomes. MiR-494 correlates with the upregulation of adhesion molecules in neutrophils of AIS patients. Systemically administered antagomiR-494 partly shifts neutrophils into the N2 phenotype in MCAO mice. AntagomiR-494-treated neutrophils exert a neuroprotective role in vitro. ChIP-seq revealed that HDAC2 targets multiple MMP genes in neutrophils of AIS patients. Further in vitro and in vivo experiments showed that antagomiR-494 repressed expression of MMP genes, including MMP7, MMP10, MMP13, and MMP16, to reduce the number of brain-infiltrating neutrophils by regulating HDAC2. Conclusion: MiR-494 may serve as an alternative predictive biomarker of the outcome of AIS patients, and antagomiR-494 treatment decreased the expression of multiple MMPs and the infiltration of neutrophils partly by targeting HDAC2.

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The Neuroprotective Effect of L-Carnitine against Glyceraldehyde-Induced Metabolic Impairment: Possible Implications in Alzheimer’s Disease

Cells ◽

10.3390/cells10082109 ◽

2021 ◽

Vol 10 (8) ◽

pp. 2109

Author(s):

Simona Magi ◽

Alessandra Preziuso ◽

Silvia Piccirillo ◽

Francesca Giampieri ◽

Danila Cianciosi ◽

...

Keyword(s):

Alzheimer’S Disease ◽

Alzheimer's Disease ◽

Cell Survival ◽

Mitochondrial Function ◽

Cortical Neurons ◽

Neurodegenerative Disorder ◽

Neuronal Damage ◽

Mitochondrial Dynamics ◽

Neuroprotective Effect ◽

Memory Loss

Alzheimer’s disease (AD) is a neurodegenerative disorder characterized by progressive cognitive regression and memory loss. Dysfunctions of both glucose metabolism and mitochondrial dynamics have been recognized as the main upstream events of the degenerative processes leading to AD. It has been recently found that correcting cell metabolism by providing alternative substrates can prevent neuronal injury by retaining mitochondrial function and reducing AD marker levels. Here, we induced an AD-like phenotype by using the glycolysis inhibitor glyceraldehyde (GA) and explored whether L-carnitine (4-N-trimethylamino-3-hydroxybutyric acid, LC) could mitigate neuronal damage, both in SH-SY5Y neuroblastoma cells and in rat primary cortical neurons. We have already reported that GA significantly modified AD marker levels; here we demonstrated that GA dramatically compromised cellular bioenergetic status, as revealed by glycolysis and oxygen consumption rate (OCR) evaluation. We found that LC ameliorated cell survival, improved OCR and ATP synthesis, prevented the loss of the mitochondrial membrane potential (Δψm) and reduced the formation of reactive oxygen species (ROS). Of note, the beneficial effect of LC did not rely on the glycolytic pathway rescue. Finally, we noticed that LC significantly reduced the increase in pTau levels induced by GA. Overall, these findings suggest that the use of LC can promote cell survival in the setting of the metabolic impairments commonly observed in AD. Our data suggest that LC may act by maintaining mitochondrial function and by reducing the pTau level.

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Role of Exendin-4 in Brain Insulin Resistance, Mitochondrial Function, and Neurite Outgrowth in Neurons under Palmitic Acid-Induced Oxidative Stress

Antioxidants ◽

10.3390/antiox10010078 ◽

2021 ◽

Vol 10 (1) ◽

pp. 78

Author(s):

Danbi Jo ◽

Gwangho Yoon ◽

Juhyun Song

Keyword(s):

Insulin Resistance ◽

Glucose Metabolism ◽

Palmitic Acid ◽

Cortical Neurons ◽

Neuronal Damage ◽

Neuronal Cell ◽

Neuronal Morphology ◽

Therapeutic Drug ◽

Signal Pathways ◽

Impaired Glucose Metabolism

Glucagon like peptide 1 (GLP-1) is an incretin hormone produced by the gut and brain, and is currently being used as a therapeutic drug for type 2 diabetes and obesity, suggesting that it regulates abnormal appetite patterns, and ameliorates impaired glucose metabolism. Many researchers have demonstrated that GLP-1 agonists and GLP-1 receptor agonists exert neuroprotective effects against brain damage. Palmitic acid (PA) is a saturated fatty acid, and increases the risk of neuroinflammation, lipotoxicity, impaired glucose metabolism, and cognitive decline. In this study, we investigated whether or not Exentin-4 (Ex-4; GLP-1 agonist) inhibits higher production of reactive oxygen species (ROS) in an SH-SY5Y neuronal cell line under PA-induced apoptosis conditions. Moreover, pre-treatment with Ex-4 in SH-SY5Y neuronal cells prevents neural apoptosis and mitochondrial dysfunction through several cellular signal pathways. In addition, insulin sensitivity in neurons is improved by Ex-4 treatment under PA-induced insulin resistance. Additionally, our imaging data showed that neuronal morphology is improved by EX-4 treatment, in spite of PA-induced neuronal damage. Furthermore, we identified that Ex-4 inhibits neuronal damage and enhanced neural complexity, such as neurite length, secondary branches, and number of neurites from soma in PA-treated SH-SY5Y. We observed that Ex-4 significantly increases neural complexity, dendritic spine morphogenesis, and development in PA treated primary cortical neurons. Hence, we suggest that GLP-1 administration may be a crucial therapeutic solution for improving neuropathology in the obese brain.

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Role of transporters and ion channels in neuronal injury under hypoxia

AJP Regulatory Integrative and Comparative Physiology ◽

10.1152/ajpregu.00528.2007 ◽

2008 ◽

Vol 294 (2) ◽

pp. R451-R457 ◽

Cited By ~ 19

Author(s):

Jin Xue ◽

Dan Zhou ◽

Hang Yao ◽

Gabriel G. Haddad

Keyword(s):

Ion Channels ◽

Cortical Neurons ◽

Cell Injury ◽

Neuronal Damage ◽

Hippocampal Slices ◽

Neuronal Injury ◽

Hypoxic Injury ◽

Significant Difference ◽

Cultured Cortical Neurons

The aims of the current study were to 1) examine the effects of hypoxia and acidosis on cultured cortical neurons and 2) explore the role of transporters and ion channels in hypoxic injury. Cell injury was measured in cultured neurons or hippocampal slices following hypoxia (1% O2) or acidosis (medium pH 6.8) treatment. Inhibitors of transporters and ion channels were employed to investigate their roles in hypoxic injury. Our results showed that 1) neuronal damage was apparent at 5–7 days of hypoxia exposure, i.e., 36–41% of total lactate dehydrogenase was released to medium and 2) acidosis alone did not lead to significant injury compared with nonacidic, normoxic controls. Pharmacological studies revealed 1) no significant difference in neuronal injury between controls (no inhibitor) and inhibition of Na+-K+-ATP pump, voltage-gated Na+ channel, ATP-sensitive K+ channel, or reverse mode of Na+/Ca2+ exchanger under hypoxia; however, 2) inhibition of NBCs with 500 μM DIDS did not cause hypoxic death in either cultured cortical neurons or hippocampal slices; 3) in contrast, inhibition of Na+/H+ exchanger isoform 1 (NHE1) with either 10 μM HOE-642 or 2 μM T-162559 resulted in dramatic hypoxic injury (+95% for HOE-642 and +100% for T-162559 relative to normoxic control, P < 0.001) on treatment day 3, when no death occurred for hypoxic controls (no inhibitor). No further damage was observed by NHE1 inhibition on treatment day 5. We conclude that inhibition of NHE1 accelerates hypoxia-induced neuronal damage. In contrast, DIDS rescues neuronal death under hypoxia. Hence, DIDS-sensitive mechanism may be a potential therapeutic target.

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tDCS confers neuroprotection by regulating LAT1-isoleucine-CBFB-PTEN signaling after rat cerebral ischemia-reperfusion injury

10.22541/au.163525012.23548218/v1 ◽

2021 ◽

Author(s):

Xujin Yao ◽

Jinyang Ren ◽

Yu Cui ◽

Songfeng Chen ◽

Jing Cheng ◽

...

Keyword(s):

Infarct Size ◽

Cortical Neurons ◽

Ischemia Reperfusion Injury ◽

Neuronal Damage ◽

Ischemia Reperfusion ◽

Glucose Deprivation ◽

Immunofluorescent Staining ◽

In Vivo Model ◽

Western Blot Assay

Background and Purpose: Isoleucine is a branched-chain amino acid serving as an essential nutrient resource and metabolic. However， its role in cerebral ischemic stroke remains unknown. Experimental Approach: Middle cerebral artery occlusion (MCAO) was used to mimic in vivo model of stroke. Oxygen-glucose deprivation insult (OGD) was used to injure cultured cortical neurons. High-Performance Liquid Chromatography (HPLC) was used to measure the level of isoleucine. A western blot assay and immunofluorescent staining were used to measure the level of CBFB and PTEN. TTC staining was used to measure the infarct size. Cell death and viability were assessed by LDH and CCK8 assays. DCS was used to stimulate cortical neurons. tDCS was used to stimulate the cortex. Key Results: Extraneuronal isoleucine is decreased and intraneuronal isoleucine is increased after rat cerebral I/R injury. Reducing intraneuronal isoleucine via inhibition of its transporter, LAT1 promotes neuronal survival whereas supplementing isoleucine aggravates neuronal damage. Isoleucine downregulates the expression of CBFB, and that acts upstream of PTEN to mediate isoleucine-induced neuronal damage after OGD insult. To identify the therapeutic approach that suppresses the ischemia-induced increase of intraneuronal isoleucine, we tested the effect of tDCS on isoleucine. Our data suggest that Cathodal tDCS can reduce cerebral infarct size. And such neuroprotection is mediated through reducing LAT1-dependent increase of intraneuronal isoleucine. Conclusions and Implications: This study identifies LAT1- dependent increase of intraneuronal isoleucine promotes neuronal death after rat cerebral I/R injury. Our results indicate that tDCS protects against rat cerebral I/R injury through regulating LAT1-isoleucine-CBFB-PTEN signaling.

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Lycopus lucidus Turcz exerts neuroprotective effect against H2O2‑induced neuroinflammation by inhibiting NLRP3 inflammasome activation in cortical neurons

10.21203/rs.3.rs-149006/v1 ◽

2021 ◽

Author(s):

Hyunseong Kim ◽

Jin Young Hong ◽

Wan-Jin Jeon ◽

Junseon Lee ◽

Seung Ho Baek ◽

...

Keyword(s):

Cell Death ◽

Nlrp3 Inflammasome ◽

Cortical Neurons ◽

Neuronal Damage ◽

Neuroprotective Effect ◽

Synaptic Connectivity ◽

Inflammasome Activation ◽

Caspase 1 ◽

Nlrp3 Inflammasome Activation ◽

Rat Cortical Neurons

Abstract Background Central nervous system (CNS) injuries are a leading cause of permanent functional impairment in humans. Nerve damage can be aggravated by neuroinflammation mediated by protein complexes known as inflammasomes, such as the NLRP3 inflammasome which is a key mediator of caspase-1 and interleukin-1β (IL-1β) /interleukin-18 (IL-18) activation. Lycopus lucidus Turcz (LLT) is a traditional medicinal herb that exerts therapeutic effects against oxidative stress, inflammation, and angiogenesis; however, it remains unclear whether LLT can directly protect neurons against damage, and the underlying molecular mechanisms are poorly understood. Methods We investigated the neuroprotective effect of LLT against hydrogen peroxide (H2O2)-induced neuronal damage in cultured primary rat cortical neurons, as well as the potential underlying mechanisms. Neuronal viability and cell death assays were used to determine the effects of LLT on neuroprotection, while the mode of cell death was confirmed using flow cytometry. Changes in the expression of inflammatory factors involved in activation of the NLRP3 inflammasome were measured using immunocytochemistry (ICC) and confirmed by real-time PCR. And, we analyzed that the effect of LLT on neurotrophic factors secretion and synaptic connectivity using ICC in H2O2-induced neuron at 7 days in vitro. Results LLT effectively protected cultured rat cortical neurons from H2O2-induced injury by significantly inhibiting NLRP3 inflammasome activation. In addition, LLT significantly reduced caspase1 activation, which is known to be induced by inflammasome formation, and consequently regulated the secretion of IL-1β/IL-18. We demonstrated that LLT enhances axonal elongation and synaptic connectivity against H2O2-induced injury of rat primary cortical neuron. Conclusions Together, these results demonstrate that LLT can directly protect cultured cortical neurons from H2O2-induced neuronal damage by inhibiting NLRP3 inflammasome activation and the secretion of caspase-1 and IL-1β/IL-18. Thus, our study provides new insights into the therapeutic mechanisms of LLT and suggests that the NLRP3 inflammasome could be a promising target for treating neurological diseases.

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Icaritin Alleviates Glutamate-Induced Neuronal Damage by Inactivating GluN2B-Containing NMDARs Through the ERK/DAPK1 Pathway

Frontiers in Neuroscience ◽

10.3389/fnins.2021.525615 ◽

2021 ◽

Vol 15 ◽

Author(s):

Song Liu ◽

Chaoming Liu ◽

Lijiao Xiong ◽

Jiali Xie ◽

Cheng Huang ◽

...

Keyword(s):

Cerebral Ischemia ◽

Cortical Neurons ◽

Neuronal Damage ◽

Glutamate Release ◽

Specific Inhibitor ◽

Expression Level ◽

Protective Mechanism ◽

Pathophysiological Mechanism ◽

Neuroprotective Effects ◽

Primary Cortical Neurons

Excitatory toxicity due to excessive glutamate release is considered the core pathophysiological mechanism of cerebral ischemia. It is primarily mediated by N-methyl-D-aspartate receptors (NMDARs) on neuronal membranes. Our previous studies have found that icaritin (ICT) exhibits neuroprotective effects against cerebral ischemia in rats, but the underlying mechanism is unclear. This study aims to investigate the protective effect of ICT on glutamate-induced neuronal injury and uncover its possible molecular mechanism. An excitatory toxicity injury model was created using rat primary cortical neurons treated with glutamate and glycine. The results showed that ICT has neuroprotective effects on glutamate-treated primary cortical neurons by increasing cell viability while reducing the rate of lactate dehydrogenase (LDH) release and reducing apoptosis. Remarkably, ICT rescued the changes in the ERK/DAPK1 signaling pathway after glutamate treatment by increasing the expression levels of p-ERK, p-DAPK1 and t-DAPK1. In addition, ICT also regulates NMDAR function during glutamate-induced injury by decreasing the expression level of the GluN2B subunit and enhancing the expression level of the GluN2A subunit. As cotreatment with the ERK-specific inhibitor U0126 and ICT abolishes the beneficial effects of ITC on the ERK/DAPK1 pathway, NMDAR subtypes and neuronal cell survival, ERK is recognized as a crucial mediator in the protective mechanism of ICT. In conclusion, our findings demonstrate that ICT has a neuroprotective effect on neuronal damage induced by glutamate, and its mechanism may be related to inactivating GluN2B-containing NMDAR through the ERK/DAPK1 pathway. This study provides a new clue for the prevention and treatment of clinical ischemic cerebrovascular diseases.

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