Background:Neuro-inflammation is regarded as one of the critical pathogenesis in neurodegenerative diseases,which is characterized by the activated microglial cells.Pectolinarin(Pec),a natural flavonoid that exists in...Background:Neuro-inflammation is regarded as one of the critical pathogenesis in neurodegenerative diseases,which is characterized by the activated microglial cells.Pectolinarin(Pec),a natural flavonoid that exists in many Chinese herbal medicines,has been reported to have various biological activities.However,the effects and mechanisms on neuro-inflammation are not clear.Methods:In this study,the inhibitory effects and mechanisms of Pec on neuro-inflammation were investigated in the LPS-stimulated microglial BV_(2) cells.BV_(2) microglial cells were treated with Pec or vehicle,followed by LPS.Enzyme-linked immunosorbent assay,real-time quantitative PCR,nitric oxide and reactive oxygen species assay,and western blot were performed to examine the effects of Pec on neuro-inflammatory responses.Results:We showed that Pec significantly inhibited the expression of tumor necrosis factorαand interleukin 6 in mRNA and protein levels induced by LPS.Moreover,the production of nitric oxide,iNOS,reactive oxygen species,and COX-2 were suppressed by Pec in LPS-stimulated microglial BV_(2) cells.In addition,Pec inhibited LPS-induced inflammation via nuclear factor kappa B signaling pathway,as evidenced by the reduction of the phosphorylation of inhibitor of nuclear factor kappa-B kinase,the degradation of IκBα,and the nuclear translocation of p65.Conclusion:Taken together,Pec exhibited anti-inflammatory effects in LPS-stimulated microglial BV_(2) cells via nuclear factor kappa B signaling pathway,which might provide therapeutic potential for neuro-inflammation and neurodegenerative diseases.展开更多
Microglia,which are the resident macrophages of the central nervous system,are an important part of the inflammatory response that occurs after cerebral ischemia.Vav guanine nucleotide exchange factor 1(Vav1) is a gua...Microglia,which are the resident macrophages of the central nervous system,are an important part of the inflammatory response that occurs after cerebral ischemia.Vav guanine nucleotide exchange factor 1(Vav1) is a guanine nucleotide exchange factor that is related to microglial activation.However,how Vav1 participates in the inflammato ry response after cerebral ischemia/reperfusion inj ury remains unclea r.In this study,we subjected rats to occlusion and repe rfusion of the middle cerebral artery and subjected the BV-2 mic roglia cell line to oxygen-glucose deprivatio n/reoxygenation to mimic cerebral ischemia/repe rfusion in vivo and in vitro,respectively.We found that Vav1 levels were increased in the brain tissue of rats subjected to occlusion and reperfusion of the middle cerebral arte ry and in BV-2 cells subjected to oxygen-glucose deprivation/reoxygenation.Silencing Vav1 reduced the cerebral infarct volume and brain water content,inhibited neuronal loss and apoptosis in the ischemic penumbra,and im p roved neurological function in rats subjected to occlusion and repe rfusion of the middle cerebral artery.Further analysis showed that Vav1 was almost exclusively localized to microglia and that Vav1 downregulation inhibited microglial activation and the NOD-like receptor pyrin 3(NLRP3) inflammasome in the ischemic penumbra,as well as the expression of inflammato ry facto rs.In addition,Vov1 knoc kdown decreased the inflammatory response exhibited by BV-2 cells after oxygen-glucose deprivation/reoxyge nation.Taken together,these findings show that silencing Vav1 attenuates inflammation and neuronal apoptosis in rats subjected to cerebral ischemia/repe rfusion through inhibiting the activation of mic roglia and NLRP3 inflammasome.展开更多
We have previously found that long-term effects of exposure to radiofrequency electromagnetic fields in 5xFAD mice with severe late-stage Alzheimer’s disease reduced both amyloid-βdeposition and glial activation,inc...We have previously found that long-term effects of exposure to radiofrequency electromagnetic fields in 5xFAD mice with severe late-stage Alzheimer’s disease reduced both amyloid-βdeposition and glial activation,including microglia.To examine whether this therapeutic effect is due to the regulation of activated microglia,we analyzed mic roglial gene expression profiles and the existence of microglia in the brain in this study.5xFAD mice at the age of 1.5 months were assigned to sham-and radiofrequency electromagnetic fields-exposed groups and then animals were exposed to 1950 MHz radiofrequency electromagnetic fields at a specific absorption rate of 5 W/kg for 2 hours/day and 5 days/week for 6 months.We conducted behavioral tests including the object recognition and Y-maze tests and molecular and histopathological analysis of amyloid precursor protein/a myloid-beta metabolism in brain tissue.We confirmed that radiofrequency electromagnetic field exposure for 6 months ameliorated cognitive impairment and amyloid-βdeposition.The expression levels of Iba1(pan-microglial marker)and colony-stimulating factor 1 receptor(CSF1R;regulates microglial prolife ration)in the hippocampus in 5xFAD mice treated with radiofrequency electromagnetic fields were significantly reduced compared with those of the sham-exposed group.Subsequently,we analyzed the expression levels of genes related to mic rogliosis and microglial function in the radiofrequency electromagnetic fields-exposed group compared to those of a CSF1R inhibitor(PLX3397)-treated group.Both radiofrequency electromagnetic fields and PLX3397 suppressed the levels of genes related to microgliosis(Csf1r,CD68,and Ccl6)and pro-inflammatory cytokine interleukin-1β.N otably,the expression levels of genes related to mic roglial function,including Trem2,Fcgr1α,Ctss,and Spi1,were decreased after long-term radiofrequency electromagnetic field exposure,which was also observed in response to microglial suppression by PLX3397.These results showed that radiofrequency electromagnetic fields ameliorated amyloid-βpathology and cognitive impairment by suppressing amyloid-βdeposition-induced microgliosis and their key regulator,CSF1R.展开更多
Epilepsy is one of the most common neurological diseases worldwide with a high prevalence and unknown pathogenesis.Further,its control is challenging.It is generally accepted that an imbalance between the excitatory a...Epilepsy is one of the most common neurological diseases worldwide with a high prevalence and unknown pathogenesis.Further,its control is challenging.It is generally accepted that an imbalance between the excitatory and inhibitory properties of the central nervous system(CNS)leads to a large number of abnormally synchronized neuronal discharges in the brain.Transient receptor potential vanilloid protein type 1(TRPV1)is a non-selective cation channel that contributes to the regulation of the nervous system and influences the excitability of the nervous system.This includes the release of neurotransmitters,action potential generation due to alterations in ion channels,synaptic transmission,and the changes in glial cells.There is abundant evidence that TRPV1 is widely expressed in the central nervous system(including microglia)and is involved in the development of epilepsy through neuroinflammation.In conclusion,microglial TRPV1 participates in neuroinflammatory reactions and functions as a potential proinflammatory mediator.This presents a novel treatment approach to regulate seizures brought on by neuroinflammation.展开更多
Ischemic stroke often induces excessive neuronal autophagy, resulting in brain damage; meanwhile, inflammatory responses stimulated by ischemia exacerbate neural injury. However, interactions between neuronal autophag...Ischemic stroke often induces excessive neuronal autophagy, resulting in brain damage; meanwhile, inflammatory responses stimulated by ischemia exacerbate neural injury. However, interactions between neuronal autophagy and microglial inflammation following ischemic stroke are poorly understood. CX3CL1/fractalkine, a membrane-bound chemokine expressed on neurons, can suppress microglial inflammation by binding to its receptor CX3CR1 on microglia. In the present study, to investigate whether autophagy could alter CX3CL1 expression on neurons and consequently change microglial inflammatory activity, middle cerebral artery occlusion(MCAO) was established in Sprague-Dawley rats to model ischemic stroke, and tissues from the ischemic penumbra were obtained to evaluate autophagy level and microglial inflammatory activity. MCAO rats were administered 3-methyladenine(autophagy inhibitor) or Tat-Beclin 1(autophagy inducer). Western blot assays were conducted to quantify expression of Beclin-1, nuclear factor kappa Bp65(NF-κB), light chain 3B(LC3B), and CX3CL1 in ischemic penumbra. Moreover, immunofluorescence staining was performed to quantify numbers of LC3B-, CX3CL1-, and Iba-1-positive cells in ischemic penumbra. In addition, enzyme linked immunosorbent assays were utilized to analyze concentrations of tumor necrosis factor alpha(TNF-α), interleukin 6(IL-6), interleukin 1 beta(IL-1β), and prostaglandin E2(PGE2). A dry/wet weight method was used to detect brain water content, while 2,3,5,-triphenyltetrazolium chloride staining was utilized to measure infarct volume. The results demonstrated that autophagy signaling(Beclin-1 and LC3B expression) in penumbra was prominently activated by MCAO, while CX3CL1 expression on autophagic neurons was significantly reduced and microglial inflammation was markedly activated. However, after inhibition of autophagy signaling with 3-methyladenine, CX3CL1 expression on neurons was obviously increased, whereas Iba-1 and NF-κB expression was downregulated; TNF-α, IL-6, IL-1β, and PGE2 levels were decreased; and cerebral edema was obviously mitigated. In contrast, after treatment with the autophagy inducer Tat-Beclin 1, CX3CL1 expression on neurons was further reduced; Iba-1 and NF-κB expression was increased; TNF-α, IL-6, IL-1β, and PGE2 levels were enhanced; and cerebral edema was aggravated. Our study suggests that ischemia-induced neuronal autophagy facilitates microglial inflammatory injury after ischemic stroke, and the efficacy of this process may be associated with downregulated CX3CL1 expression on autophagic neurons.展开更多
Interleukin-4 plays an important protective role in Alzheimer’s disease by regulating microglial phenotype,phagocytosis of amyloid-β,and secretion of anti-inflammatory and neurotrophic cytokines.Recently,increasing ...Interleukin-4 plays an important protective role in Alzheimer’s disease by regulating microglial phenotype,phagocytosis of amyloid-β,and secretion of anti-inflammatory and neurotrophic cytokines.Recently,increasing evidence has suggested that autophagy regulates innate immunity by affecting M1/M2 polarization of microglia/macrophages.However,the role of interleukin-4 in microglial autophagy is unknown.In view of this,BV2 microglia were treated with 0,10,20 or 50 ng/mL interleukin-4 for 24,48,or 72 hours.Subsequently,light chain 3-II and p62 protein expression levels were detected by western blot assay.BV2 microglia were incubated with interleukin-4(20 ng/mL,experimental group),3-methyladenine(500μM,autophagy inhibitor,negative control group),rapamycin(100 nM,autophagy inductor,positive control group),3-methyladenine+interleukin-4(rescue group),or without treatment for 24 hours,and then exposed to amyloid-β(1μM,model group)or vehicle control(control)for 24 hours.LC3-II and p62 protein expression levels were again detected by western blot assay.In addition,expression levels of multiple markers of M1 and M2 phenotype were assessed by real-time fluorescence quantitative polymerase chain reaction,while intracellular and supernatant amyloid-βprotein levels were measured by enzyme-linked immunosorbent assay.Our results showed that interleukin-4 induced microglial autophagic flux,most significantly at 20 ng/mL for 48 hours.Interleukin-4 pretreated microglia inhibited blockade of amyloid-β-induced autophagic flux,and promoted amyloid-βuptake and degradation partly through autophagic flux,but inhibited switching of amyloid-β-induced M1 phenotype independent on autophagic flux.These results indicate that interleukin-4 pretreated microglia increases uptake and degradation of amyloid-βin a process partly mediated by autophagy,which may play a protective role against Alzheimer’s disease.展开更多
Traumatic brain injury(TBI)remains one of the leading causes of disability and death in infants and children.Studies have demonstrated that the youngest age group(especially≤4 years old)exhibit worse functional outco...Traumatic brain injury(TBI)remains one of the leading causes of disability and death in infants and children.Studies have demonstrated that the youngest age group(especially≤4 years old)exhibit worse functional outcome following moderate to severe TBI compared to older children or adults(Anderson et al.,2005;Emami et al.,2017).These data suggest that age-at-injury may be an important determinant of outcome,as damage to the developing brain at a young age likely disrupts展开更多
Objective: To determine the anti-neuroinflammatory activity of Moringa oleifera leaf extract(MLE) under lipopolysaccharide stimulation of mouse murine microglia BV2 cells in vitro. Methods: The cytotoxicity effect of ...Objective: To determine the anti-neuroinflammatory activity of Moringa oleifera leaf extract(MLE) under lipopolysaccharide stimulation of mouse murine microglia BV2 cells in vitro. Methods: The cytotoxicity effect of MLE was investigated by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide assay. The inflammatory response of BV-2 cells were induced with lipopolysaccharide. The generation of nitric oxide levels was determined by using Griess assay and the level of pro-inflammatory cytokines(IL-1β, IL-6 and TNF-α) was evaluated by ELISA kit. The expression of iNOS, COX-2 as well as IκB-ααwas carried out by immunoblot analysis. Results: MLE reduced the nitric oxide production in concentration-dependent manner, and maintained the viability of BV-2 microglial cells which indicated absence of toxicity. In addition, MLE repressed the activation of nuclear factor kappa B by arresting the deterioration of IκB-α, consequently resulted in suppression of cytokines expression such as COX-2 and iNOS. Conclusions: MLE inhibitory activities are associated with the inhibition of nuclear factor kappa B transcriptional activity in BV2 microglial cells. Thus MLE may offer a substantial treatment for neuroinflammatory diseases.展开更多
Traumatic brain injury(TBI)results in a range of neuroinflammatory events that vary depending on the type and extent of injury.Central to this is the activation of tissue resident microglia and infiltration of periphe...Traumatic brain injury(TBI)results in a range of neuroinflammatory events that vary depending on the type and extent of injury.Central to this is the activation of tissue resident microglia and infiltration of peripheral macrophages,which phagocytose debris and/or secrete a range of cytokines,chemokines and other factors which modify the injured environment to展开更多
Microglial cells are the key innate immune cells in the brain and they are crucial in maintaining brain parenchyma homeostasis.Under physiological conditions,microglial cells assume a ramified morphology with a small ...Microglial cells are the key innate immune cells in the brain and they are crucial in maintaining brain parenchyma homeostasis.Under physiological conditions,microglial cells assume a ramified morphology with a small cell body and an extensive network of fine processes,which secrete neurotrophic factors and patrol the surroundings in search for pathogens and eliminate cellular debris via phagocytosis.Microglial cells express a repertoire of pattern recognition receptors(PRRs)that enable them to detect diverse danger-associated molecular patterns(DAMPs)released from damaged cells or cells under stress,or pathogen-associated molecular patterns generated by pathogens during infection.展开更多
Background:Inflammation and damage to neurons and other cells in the nervous system can cause disorders of the central nervous system.Microglial cells are activated by pathogen infection and injury to release nitric o...Background:Inflammation and damage to neurons and other cells in the nervous system can cause disorders of the central nervous system.Microglial cells are activated by pathogen infection and injury to release nitric oxide.Valerian(Valeriana officinalis)has been used as a sedative for the treatment of neurological diseases.This study evaluated inflammation of microglial cells and tumor necrosis factorαand induced nitric oxide synthetase gene expression influenced by valerian extract.Methods:Microglial cells were isolated from mice.Lipopolysaccharide(1 ng/mL)was used to induce inflammation and nitric oxide production in cells for an hour.The inflamed cells were then treated with different concentrations(0.1,0.5,2.5,20,and 50μL/mL)of valerian alcoholic extract for 1 and 24 h.nitric oxide production and tumor necrosis factorαand induced nitric oxide synthetase gene expression were determine by Griess assay and real-time polymerase chain reaction,respectively.Results:The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay showed no toxicity in several concentrations of the valerian extract.In addition,concentrations from 0.1 to 2.5μL/mL significantly reduced inflammation and nitric oxide production in mouse microglial cells to levels observed in control samples.Furthermore,tumor necrosis factorαand induced nitric oxide synthetase gene expression decreased when 2.5μL/mL extract was used.Conclusion:Based on these results,it can be concluded that 2.5μL/mL valerian alcoholic extract is effective as a candidate alternative medicine for reducing inflammation and nitric oxide production and consequently,the inflammatory symptoms of neurodegeneration.展开更多
Microglial activation plays an important role in a panel of neurological disorders such as multiple sclerosis(MS) and Parkinson's disease(PD),and is a key target for developing therapeutic strategies for these dis...Microglial activation plays an important role in a panel of neurological disorders such as multiple sclerosis(MS) and Parkinson's disease(PD),and is a key target for developing therapeutic strategies for these diseases.Ketogenic diet (KD),which is able to inhibit microglial activation in substantia nigra pars compacta of mice,has been shown effective in a mouse model of PD,possibly through increasing D-β-hydroxybutyrate(D-β-HB),a major component of ketone bodies.To verify this,we developed an in vitro model of microglia activation with a microglia line,BV-2,and investigated how D-β-HB have an effect on the LPS-stimulated BV-2 cells.We found D-β-HB is able to recover the cell viability,and inhibit the production of inflammatory mediators and cytokines such as ROS,nitrite,IL-1β,TNF-α,and IL-6,which otherwise were increased in LPS-stimulated BV-2 cells.We conclude that the LPS induced BV-2 cells activation is a valid in vitro model of microglia activation.D-β-HB is able to suppress the activation of BV-2 cells, which might account for one of the possible reasons of KD therapy on the PD model.展开更多
Glaucoma is a multifactorial optic neuropathy characterized by the damage and death of the retinal ganglion cells.This disease results in vision loss and blindness.Any vision loss resulting from the disease cannot be ...Glaucoma is a multifactorial optic neuropathy characterized by the damage and death of the retinal ganglion cells.This disease results in vision loss and blindness.Any vision loss resulting from the disease cannot be restored and nowadays there is no available cure for glaucoma; however an early detection and treatment,could offer neuronal protection and avoid later serious damages to the visual function.A full understanding of the etiology of the disease will still require the contribution of many scientific efforts.Glial activation has been observed in glaucoma,being microglial proliferation a hallmark in this neurodegenerative disease.A typical project studying these cellular changes involved in glaucoma often needs thousands of images- from several animals- covering different layers and regions of the retina.The gold standard to evaluate them is the manual count.This method requires a large amount of time from specialized personnel.It is a tedious process and prone to human error.We present here a new method to count microglial cells by using a computer algorithm.It counts in one hour the same number of images that a researcher counts in four weeks,with no loss of reliability.展开更多
A major challenge for the efficient treatment of traumatic brain injury is the need for therapeutic molecules to cross the blood-brain barrier to enter and accumulate in brain tissue.To overcome this problem,researche...A major challenge for the efficient treatment of traumatic brain injury is the need for therapeutic molecules to cross the blood-brain barrier to enter and accumulate in brain tissue.To overcome this problem,researchers have begun to focus on nanocarriers and other brain-targeting drug delivery systems.In this review,we summarize the epidemiology,basic pathophysiology,current clinical treatment,the establishment of models,and the evaluation indicators that are commonly used for traumatic brain injury.We also report the current status of traumatic brain injury when treated with nanocarriers such as liposomes and vesicles.Nanocarriers can overcome a variety of key biological barriers,improve drug bioavailability,increase intracellular penetration and retention time,achieve drug enrichment,control drug release,and achieve brain-targeting drug delivery.However,the application of nanocarriers remains in the basic research stage and has yet to be fully translated to the clinic.展开更多
Microglia play multiple roles in such processes as brain development,homeostasis,and pathology.Due to their diverse mechanisms of functions,the complex sub-classifications,and the large differences between different s...Microglia play multiple roles in such processes as brain development,homeostasis,and pathology.Due to their diverse mechanisms of functions,the complex sub-classifications,and the large differences between different species,especially compared with humans,very different or even opposite conclusions can be drawn from studies with different research models.The choice of appropriate research models and the associated tools are thus key ingredients of studies on microglia.Mice are the most commonly used animal models.In this review,we summarize in vitro and in vivo models of mouse and human-derived microglial research models,including microglial cell lines,primary microglia,induced microglia-like cells,transgenic mice,human-mouse chimeric models,and microglial replacement models.We also summarize recent developments in novel single-cell and in vivo imaging technologies.We hope our review can serve as an efficient reference for the future study of microglia.展开更多
Epilepsy frequently leads to cognitive dysfunction and approaches to treatment remain limited.Although regular exercise effectively improves learning and memory functions across multiple neurological diseases,its appl...Epilepsy frequently leads to cognitive dysfunction and approaches to treatment remain limited.Although regular exercise effectively improves learning and memory functions across multiple neurological diseases,its application in patients with epilepsy remains controversial.Here,we adopted a 14-day treadmill-exercise paradigm in a pilocarpine injection-induced mouse model of epilepsy.Cognitive assays confirmed the improvement of object and spatial memory after endurance training,and electrophysiological studies revealed the maintenance of hippocampal plasticity as a result of physical exercise.Investigations of the mechanisms underlying this effect revealed that exercise protected parvalbumin interneurons,probably via the suppression of neuroinflammation and improved integrity of blood-brain barrier.In summary,this work identified a previously unknown mechanism through which exercise improves cognitive rehabilitation in epilepsy.展开更多
Inflammation plays important roles in the progress of neurodegenerative diseases,such as Parkinson’s disease and Alzheimer’s disease.Microglia is responsible for the homeostasis of the central nervous system(CNS),an...Inflammation plays important roles in the progress of neurodegenerative diseases,such as Parkinson’s disease and Alzheimer’s disease.Microglia is responsible for the homeostasis of the central nervous system(CNS),and involved in the neuroinflammation.Therefore,it could be potential in treatment of neurodegenerative diseases to suppress the microglia-mediated neuroinflammation.Mangiferin,a major glucoside of xanthone in Anemarrhena Rhizome,has anti-inflammatory,anti-diabetes,and anti-oxidative properties.However,the effect of mangiferin on the inflammatary responses of microglia cells are still poorly understand.In this study,we investigated the mechanism by which mangiferin inhibited inflammation in LPS-induced BV2 microglia cells.BV2 cells were pretreatment with mangiferin followed by LPS stimulation.In vitro assays,NO and cytokines production were quantified.Western blot and immunocytochemistry were used to examine the effect of mangiferin on the polarization of BV2 cells and signaling pathway.The results showed that mangiferin treatment significantly reduced NO,IL-1β,IL-6 and TNF-αproduction,also reduced the mRNA and protein of iNOS and COX-2,promoted the polarization of inflammatory toward anti-inflammatory,and inhibited activation of NF-κB and NLRP3 inflammasome.These data suggest that mangiferin has an anti-neuroinflammatory property via regulating microglia macrophage polarization and suppressing NF-κB and NLRP3 signaling pathway,and may act as a potential natural therapeutic candidate for neuroinflammatory diseases.展开更多
Background:Neuron-microglia communication plays a crucial role in the motor neurons(MNs)death in amyotrophic lateral sclerosis(ALS).Neurons can express chemokine(C-X3-C motif)ligand 1(CX3CL1),which mediates microglial...Background:Neuron-microglia communication plays a crucial role in the motor neurons(MNs)death in amyotrophic lateral sclerosis(ALS).Neurons can express chemokine(C-X3-C motif)ligand 1(CX3CL1),which mediates microglial activation via interacting with its sole receptor CX3CR1 in microglia.In the present study,we aimed to investigate the dynamic changes of CX3CL1/CX3CR1 axis during microglial activation and MNs loss in SOD1G93A mouse model of ALS.Methods:qPCR,western blot and immunofluorescent staining were used to examine the mRNA and protein levels and localization of CX3CL1/CX3CR1 in the anterior horn region of spinal cord in both SOD1G93A mice and their agematched wild type(WT)littermates at 40,60,90 and 120 days of age.The M1/M2 microglial activation in the spinal cord tissues of SOD1G93A mice and WT mice were evaluated by immunofluorescent staining of M1/M2 markers and further confirmed by qPCR analysis of M1/M2-related cytokines.Results:The immunofluorescent staining revealed that CX3CL1 was predominately expressed in MNs,while CX3CR1 was highly expressed in microglia in the anterior horn region of spinal cord.Compared with age-matched WT mice,CX3CL1 mRNA level was elevated at 40 days but decreased at 90 and 120 days in the anterior horn region of spinal cords in ALS mice.Consistently,CX3CR1 mRNA level was increased at 90 and 120 days.Western blot assay further confirmed the dynamic changes of CX3CL1/CX3CR1 axis in ALS mice.Additionally,the levels of M1/M2 markers of microglia and their related cytokines in the anterior horn region of spinal cord in ALS mice were increased at 90 and 120 days.Moreover,while M1-related cytokines in ALS mice were persistently increased at 120 days,the upregulated M2-related cytokines started to decline at 120 days,suggesting an altered microglial activation.Conclusions:Our data revealed the dynamic changes of CX3CL1/CX3CR1 axis and an imbalanced M1/M2 microglial activation during ALS pathological progression.These findings may help identify potential molecular targets for ALS therapy.展开更多
基金supported by the grants from the Training Program for Young Backbone Teachers in Colleges and Universities of Henan Province(2021GGJS103)the Program for Science and Technology Innovation Team in Universities of Henan Province(22IRTSTHN030)the Henan Neural Development Engineering Research Center for Children Foundation(SG201909).
文摘Background:Neuro-inflammation is regarded as one of the critical pathogenesis in neurodegenerative diseases,which is characterized by the activated microglial cells.Pectolinarin(Pec),a natural flavonoid that exists in many Chinese herbal medicines,has been reported to have various biological activities.However,the effects and mechanisms on neuro-inflammation are not clear.Methods:In this study,the inhibitory effects and mechanisms of Pec on neuro-inflammation were investigated in the LPS-stimulated microglial BV_(2) cells.BV_(2) microglial cells were treated with Pec or vehicle,followed by LPS.Enzyme-linked immunosorbent assay,real-time quantitative PCR,nitric oxide and reactive oxygen species assay,and western blot were performed to examine the effects of Pec on neuro-inflammatory responses.Results:We showed that Pec significantly inhibited the expression of tumor necrosis factorαand interleukin 6 in mRNA and protein levels induced by LPS.Moreover,the production of nitric oxide,iNOS,reactive oxygen species,and COX-2 were suppressed by Pec in LPS-stimulated microglial BV_(2) cells.In addition,Pec inhibited LPS-induced inflammation via nuclear factor kappa B signaling pathway,as evidenced by the reduction of the phosphorylation of inhibitor of nuclear factor kappa-B kinase,the degradation of IκBα,and the nuclear translocation of p65.Conclusion:Taken together,Pec exhibited anti-inflammatory effects in LPS-stimulated microglial BV_(2) cells via nuclear factor kappa B signaling pathway,which might provide therapeutic potential for neuro-inflammation and neurodegenerative diseases.
基金Natural Science Foundation of Liaoning Province (General Program),No.2017010825 (to JQ)。
文摘Microglia,which are the resident macrophages of the central nervous system,are an important part of the inflammatory response that occurs after cerebral ischemia.Vav guanine nucleotide exchange factor 1(Vav1) is a guanine nucleotide exchange factor that is related to microglial activation.However,how Vav1 participates in the inflammato ry response after cerebral ischemia/reperfusion inj ury remains unclea r.In this study,we subjected rats to occlusion and repe rfusion of the middle cerebral artery and subjected the BV-2 mic roglia cell line to oxygen-glucose deprivatio n/reoxygenation to mimic cerebral ischemia/repe rfusion in vivo and in vitro,respectively.We found that Vav1 levels were increased in the brain tissue of rats subjected to occlusion and reperfusion of the middle cerebral arte ry and in BV-2 cells subjected to oxygen-glucose deprivation/reoxygenation.Silencing Vav1 reduced the cerebral infarct volume and brain water content,inhibited neuronal loss and apoptosis in the ischemic penumbra,and im p roved neurological function in rats subjected to occlusion and repe rfusion of the middle cerebral artery.Further analysis showed that Vav1 was almost exclusively localized to microglia and that Vav1 downregulation inhibited microglial activation and the NOD-like receptor pyrin 3(NLRP3) inflammasome in the ischemic penumbra,as well as the expression of inflammato ry facto rs.In addition,Vov1 knoc kdown decreased the inflammatory response exhibited by BV-2 cells after oxygen-glucose deprivation/reoxyge nation.Taken together,these findings show that silencing Vav1 attenuates inflammation and neuronal apoptosis in rats subjected to cerebral ischemia/repe rfusion through inhibiting the activation of mic roglia and NLRP3 inflammasome.
基金Institute of Information&Communications Technology Planning&Evaluation(IITP)grant funded by Korea government(MSIT),Nos.2017-0-00961 and 2019-0-00102(to HDC)。
文摘We have previously found that long-term effects of exposure to radiofrequency electromagnetic fields in 5xFAD mice with severe late-stage Alzheimer’s disease reduced both amyloid-βdeposition and glial activation,including microglia.To examine whether this therapeutic effect is due to the regulation of activated microglia,we analyzed mic roglial gene expression profiles and the existence of microglia in the brain in this study.5xFAD mice at the age of 1.5 months were assigned to sham-and radiofrequency electromagnetic fields-exposed groups and then animals were exposed to 1950 MHz radiofrequency electromagnetic fields at a specific absorption rate of 5 W/kg for 2 hours/day and 5 days/week for 6 months.We conducted behavioral tests including the object recognition and Y-maze tests and molecular and histopathological analysis of amyloid precursor protein/a myloid-beta metabolism in brain tissue.We confirmed that radiofrequency electromagnetic field exposure for 6 months ameliorated cognitive impairment and amyloid-βdeposition.The expression levels of Iba1(pan-microglial marker)and colony-stimulating factor 1 receptor(CSF1R;regulates microglial prolife ration)in the hippocampus in 5xFAD mice treated with radiofrequency electromagnetic fields were significantly reduced compared with those of the sham-exposed group.Subsequently,we analyzed the expression levels of genes related to mic rogliosis and microglial function in the radiofrequency electromagnetic fields-exposed group compared to those of a CSF1R inhibitor(PLX3397)-treated group.Both radiofrequency electromagnetic fields and PLX3397 suppressed the levels of genes related to microgliosis(Csf1r,CD68,and Ccl6)and pro-inflammatory cytokine interleukin-1β.N otably,the expression levels of genes related to mic roglial function,including Trem2,Fcgr1α,Ctss,and Spi1,were decreased after long-term radiofrequency electromagnetic field exposure,which was also observed in response to microglial suppression by PLX3397.These results showed that radiofrequency electromagnetic fields ameliorated amyloid-βpathology and cognitive impairment by suppressing amyloid-βdeposition-induced microgliosis and their key regulator,CSF1R.
文摘Epilepsy is one of the most common neurological diseases worldwide with a high prevalence and unknown pathogenesis.Further,its control is challenging.It is generally accepted that an imbalance between the excitatory and inhibitory properties of the central nervous system(CNS)leads to a large number of abnormally synchronized neuronal discharges in the brain.Transient receptor potential vanilloid protein type 1(TRPV1)is a non-selective cation channel that contributes to the regulation of the nervous system and influences the excitability of the nervous system.This includes the release of neurotransmitters,action potential generation due to alterations in ion channels,synaptic transmission,and the changes in glial cells.There is abundant evidence that TRPV1 is widely expressed in the central nervous system(including microglia)and is involved in the development of epilepsy through neuroinflammation.In conclusion,microglial TRPV1 participates in neuroinflammatory reactions and functions as a potential proinflammatory mediator.This presents a novel treatment approach to regulate seizures brought on by neuroinflammation.
基金supported by the National Natural Science Foundation of China,No.81660383(to YHD),81860411(to HYH)a grant from the Applied Basic Research Projects of Yunnan Province of China,No.2017FB113(to YHD)the Scientific Research Fund of Yunnan Provincial Department of Education of China,No.2018JS016(to HYH)
文摘Ischemic stroke often induces excessive neuronal autophagy, resulting in brain damage; meanwhile, inflammatory responses stimulated by ischemia exacerbate neural injury. However, interactions between neuronal autophagy and microglial inflammation following ischemic stroke are poorly understood. CX3CL1/fractalkine, a membrane-bound chemokine expressed on neurons, can suppress microglial inflammation by binding to its receptor CX3CR1 on microglia. In the present study, to investigate whether autophagy could alter CX3CL1 expression on neurons and consequently change microglial inflammatory activity, middle cerebral artery occlusion(MCAO) was established in Sprague-Dawley rats to model ischemic stroke, and tissues from the ischemic penumbra were obtained to evaluate autophagy level and microglial inflammatory activity. MCAO rats were administered 3-methyladenine(autophagy inhibitor) or Tat-Beclin 1(autophagy inducer). Western blot assays were conducted to quantify expression of Beclin-1, nuclear factor kappa Bp65(NF-κB), light chain 3B(LC3B), and CX3CL1 in ischemic penumbra. Moreover, immunofluorescence staining was performed to quantify numbers of LC3B-, CX3CL1-, and Iba-1-positive cells in ischemic penumbra. In addition, enzyme linked immunosorbent assays were utilized to analyze concentrations of tumor necrosis factor alpha(TNF-α), interleukin 6(IL-6), interleukin 1 beta(IL-1β), and prostaglandin E2(PGE2). A dry/wet weight method was used to detect brain water content, while 2,3,5,-triphenyltetrazolium chloride staining was utilized to measure infarct volume. The results demonstrated that autophagy signaling(Beclin-1 and LC3B expression) in penumbra was prominently activated by MCAO, while CX3CL1 expression on autophagic neurons was significantly reduced and microglial inflammation was markedly activated. However, after inhibition of autophagy signaling with 3-methyladenine, CX3CL1 expression on neurons was obviously increased, whereas Iba-1 and NF-κB expression was downregulated; TNF-α, IL-6, IL-1β, and PGE2 levels were decreased; and cerebral edema was obviously mitigated. In contrast, after treatment with the autophagy inducer Tat-Beclin 1, CX3CL1 expression on neurons was further reduced; Iba-1 and NF-κB expression was increased; TNF-α, IL-6, IL-1β, and PGE2 levels were enhanced; and cerebral edema was aggravated. Our study suggests that ischemia-induced neuronal autophagy facilitates microglial inflammatory injury after ischemic stroke, and the efficacy of this process may be associated with downregulated CX3CL1 expression on autophagic neurons.
基金supported by the Natural Science Foundation of Liaoning Province of China,No.20170541036(to HYL)
文摘Interleukin-4 plays an important protective role in Alzheimer’s disease by regulating microglial phenotype,phagocytosis of amyloid-β,and secretion of anti-inflammatory and neurotrophic cytokines.Recently,increasing evidence has suggested that autophagy regulates innate immunity by affecting M1/M2 polarization of microglia/macrophages.However,the role of interleukin-4 in microglial autophagy is unknown.In view of this,BV2 microglia were treated with 0,10,20 or 50 ng/mL interleukin-4 for 24,48,or 72 hours.Subsequently,light chain 3-II and p62 protein expression levels were detected by western blot assay.BV2 microglia were incubated with interleukin-4(20 ng/mL,experimental group),3-methyladenine(500μM,autophagy inhibitor,negative control group),rapamycin(100 nM,autophagy inductor,positive control group),3-methyladenine+interleukin-4(rescue group),or without treatment for 24 hours,and then exposed to amyloid-β(1μM,model group)or vehicle control(control)for 24 hours.LC3-II and p62 protein expression levels were again detected by western blot assay.In addition,expression levels of multiple markers of M1 and M2 phenotype were assessed by real-time fluorescence quantitative polymerase chain reaction,while intracellular and supernatant amyloid-βprotein levels were measured by enzyme-linked immunosorbent assay.Our results showed that interleukin-4 induced microglial autophagic flux,most significantly at 20 ng/mL for 48 hours.Interleukin-4 pretreated microglia inhibited blockade of amyloid-β-induced autophagic flux,and promoted amyloid-βuptake and degradation partly through autophagic flux,but inhibited switching of amyloid-β-induced M1 phenotype independent on autophagic flux.These results indicate that interleukin-4 pretreated microglia increases uptake and degradation of amyloid-βin a process partly mediated by autophagy,which may play a protective role against Alzheimer’s disease.
文摘Traumatic brain injury(TBI)remains one of the leading causes of disability and death in infants and children.Studies have demonstrated that the youngest age group(especially≤4 years old)exhibit worse functional outcome following moderate to severe TBI compared to older children or adults(Anderson et al.,2005;Emami et al.,2017).These data suggest that age-at-injury may be an important determinant of outcome,as damage to the developing brain at a young age likely disrupts
基金supported by the Basic Science Research Program through the National Research Foundation of Korea(NRF) funded by the Ministry of Science and ICT(NRF-2017R1C1B2010276 and 2017R1A2A2A07001035)
文摘Objective: To determine the anti-neuroinflammatory activity of Moringa oleifera leaf extract(MLE) under lipopolysaccharide stimulation of mouse murine microglia BV2 cells in vitro. Methods: The cytotoxicity effect of MLE was investigated by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide assay. The inflammatory response of BV-2 cells were induced with lipopolysaccharide. The generation of nitric oxide levels was determined by using Griess assay and the level of pro-inflammatory cytokines(IL-1β, IL-6 and TNF-α) was evaluated by ELISA kit. The expression of iNOS, COX-2 as well as IκB-ααwas carried out by immunoblot analysis. Results: MLE reduced the nitric oxide production in concentration-dependent manner, and maintained the viability of BV-2 microglial cells which indicated absence of toxicity. In addition, MLE repressed the activation of nuclear factor kappa B by arresting the deterioration of IκB-α, consequently resulted in suppression of cytokines expression such as COX-2 and iNOS. Conclusions: MLE inhibitory activities are associated with the inhibition of nuclear factor kappa B transcriptional activity in BV2 microglial cells. Thus MLE may offer a substantial treatment for neuroinflammatory diseases.
基金supported by the National Health and Medical Research Council of AustraliaProject grant 1045125 and Fellowship(to AMT)628344support from Stem Cells Australia
文摘Traumatic brain injury(TBI)results in a range of neuroinflammatory events that vary depending on the type and extent of injury.Central to this is the activation of tissue resident microglia and infiltration of peripheral macrophages,which phagocytose debris and/or secrete a range of cytokines,chemokines and other factors which modify the injured environment to
基金supported in part by grants from the Disciplinary Group of Psychology and Neuroscience Xinxiang Medical University(2016PN-KFKT-06)a visiting professorship from University of Tours(to LHJ)
文摘Microglial cells are the key innate immune cells in the brain and they are crucial in maintaining brain parenchyma homeostasis.Under physiological conditions,microglial cells assume a ramified morphology with a small cell body and an extensive network of fine processes,which secrete neurotrophic factors and patrol the surroundings in search for pathogens and eliminate cellular debris via phagocytosis.Microglial cells express a repertoire of pattern recognition receptors(PRRs)that enable them to detect diverse danger-associated molecular patterns(DAMPs)released from damaged cells or cells under stress,or pathogen-associated molecular patterns generated by pathogens during infection.
文摘Background:Inflammation and damage to neurons and other cells in the nervous system can cause disorders of the central nervous system.Microglial cells are activated by pathogen infection and injury to release nitric oxide.Valerian(Valeriana officinalis)has been used as a sedative for the treatment of neurological diseases.This study evaluated inflammation of microglial cells and tumor necrosis factorαand induced nitric oxide synthetase gene expression influenced by valerian extract.Methods:Microglial cells were isolated from mice.Lipopolysaccharide(1 ng/mL)was used to induce inflammation and nitric oxide production in cells for an hour.The inflamed cells were then treated with different concentrations(0.1,0.5,2.5,20,and 50μL/mL)of valerian alcoholic extract for 1 and 24 h.nitric oxide production and tumor necrosis factorαand induced nitric oxide synthetase gene expression were determine by Griess assay and real-time polymerase chain reaction,respectively.Results:The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay showed no toxicity in several concentrations of the valerian extract.In addition,concentrations from 0.1 to 2.5μL/mL significantly reduced inflammation and nitric oxide production in mouse microglial cells to levels observed in control samples.Furthermore,tumor necrosis factorαand induced nitric oxide synthetase gene expression decreased when 2.5μL/mL extract was used.Conclusion:Based on these results,it can be concluded that 2.5μL/mL valerian alcoholic extract is effective as a candidate alternative medicine for reducing inflammation and nitric oxide production and consequently,the inflammatory symptoms of neurodegeneration.
文摘Microglial activation plays an important role in a panel of neurological disorders such as multiple sclerosis(MS) and Parkinson's disease(PD),and is a key target for developing therapeutic strategies for these diseases.Ketogenic diet (KD),which is able to inhibit microglial activation in substantia nigra pars compacta of mice,has been shown effective in a mouse model of PD,possibly through increasing D-β-hydroxybutyrate(D-β-HB),a major component of ketone bodies.To verify this,we developed an in vitro model of microglia activation with a microglia line,BV-2,and investigated how D-β-HB have an effect on the LPS-stimulated BV-2 cells.We found D-β-HB is able to recover the cell viability,and inhibit the production of inflammatory mediators and cytokines such as ROS,nitrite,IL-1β,TNF-α,and IL-6,which otherwise were increased in LPS-stimulated BV-2 cells.We conclude that the LPS induced BV-2 cells activation is a valid in vitro model of microglia activation.D-β-HB is able to suppress the activation of BV-2 cells, which might account for one of the possible reasons of KD therapy on the PD model.
基金supported by the Science Foundation of Arizona through the Bisgrove Program to PdG,Grant Number:BSP 0529-13the Ophthalmological Network OFTARED(RD12-0034/0002)+5 种基金the Institute of Health Carlos IIIthe PN I+D+i 2008–2011the ISCIII-Subdireccion General de Redes y Centros de Investigación Cooperativathe European Programme FEDERthe project SAF2014-53779-Rthe project:“The role of encapsulated NSAIDs in PLGA microparticles as a neuroprotective treatment” funded by the Spanish Ministry of Economy and Competitiveness
文摘Glaucoma is a multifactorial optic neuropathy characterized by the damage and death of the retinal ganglion cells.This disease results in vision loss and blindness.Any vision loss resulting from the disease cannot be restored and nowadays there is no available cure for glaucoma; however an early detection and treatment,could offer neuronal protection and avoid later serious damages to the visual function.A full understanding of the etiology of the disease will still require the contribution of many scientific efforts.Glial activation has been observed in glaucoma,being microglial proliferation a hallmark in this neurodegenerative disease.A typical project studying these cellular changes involved in glaucoma often needs thousands of images- from several animals- covering different layers and regions of the retina.The gold standard to evaluate them is the manual count.This method requires a large amount of time from specialized personnel.It is a tedious process and prone to human error.We present here a new method to count microglial cells by using a computer algorithm.It counts in one hour the same number of images that a researcher counts in four weeks,with no loss of reliability.
基金supported by the Natural Science Foundation of Beijing,No.L222126(to LD)。
文摘A major challenge for the efficient treatment of traumatic brain injury is the need for therapeutic molecules to cross the blood-brain barrier to enter and accumulate in brain tissue.To overcome this problem,researchers have begun to focus on nanocarriers and other brain-targeting drug delivery systems.In this review,we summarize the epidemiology,basic pathophysiology,current clinical treatment,the establishment of models,and the evaluation indicators that are commonly used for traumatic brain injury.We also report the current status of traumatic brain injury when treated with nanocarriers such as liposomes and vesicles.Nanocarriers can overcome a variety of key biological barriers,improve drug bioavailability,increase intracellular penetration and retention time,achieve drug enrichment,control drug release,and achieve brain-targeting drug delivery.However,the application of nanocarriers remains in the basic research stage and has yet to be fully translated to the clinic.
基金the National Key Research and Development Program of China(2017YFC0909200)the National Natural Science Foundation of China(81671336)+1 种基金Shanghai Key Laboratory of Psychotic Disorders(YG2016ZD06)the Shanghai Mental Health Center(2019-YJ06).
文摘Microglia play multiple roles in such processes as brain development,homeostasis,and pathology.Due to their diverse mechanisms of functions,the complex sub-classifications,and the large differences between different species,especially compared with humans,very different or even opposite conclusions can be drawn from studies with different research models.The choice of appropriate research models and the associated tools are thus key ingredients of studies on microglia.Mice are the most commonly used animal models.In this review,we summarize in vitro and in vivo models of mouse and human-derived microglial research models,including microglial cell lines,primary microglia,induced microglia-like cells,transgenic mice,human-mouse chimeric models,and microglial replacement models.We also summarize recent developments in novel single-cell and in vivo imaging technologies.We hope our review can serve as an efficient reference for the future study of microglia.
基金supported by STI2030-Major Projects,No.2022ZD0207600 (to LZ)the National Natural Science Foundation of China,Nos.821 71446 (to JY),U22A20301 (to KFS),32070955 (to LZ)+1 种基金Guangdong Basic and Applied Basic Research Foundation,No.202381515040015 (to LZ)Science and Technology Program of Guangzhou of China,No.202007030012 (to KFS and LZ)
文摘Epilepsy frequently leads to cognitive dysfunction and approaches to treatment remain limited.Although regular exercise effectively improves learning and memory functions across multiple neurological diseases,its application in patients with epilepsy remains controversial.Here,we adopted a 14-day treadmill-exercise paradigm in a pilocarpine injection-induced mouse model of epilepsy.Cognitive assays confirmed the improvement of object and spatial memory after endurance training,and electrophysiological studies revealed the maintenance of hippocampal plasticity as a result of physical exercise.Investigations of the mechanisms underlying this effect revealed that exercise protected parvalbumin interneurons,probably via the suppression of neuroinflammation and improved integrity of blood-brain barrier.In summary,this work identified a previously unknown mechanism through which exercise improves cognitive rehabilitation in epilepsy.
基金the National Natural Science Foundation of China(No.81603357)the Natural Science Basic Research Plan in Shaanxi Province of China(No.2017.Q8043)the Scientific Research Program Funded by Shaanxi Provincial Education Department(No.20JY011).
文摘Inflammation plays important roles in the progress of neurodegenerative diseases,such as Parkinson’s disease and Alzheimer’s disease.Microglia is responsible for the homeostasis of the central nervous system(CNS),and involved in the neuroinflammation.Therefore,it could be potential in treatment of neurodegenerative diseases to suppress the microglia-mediated neuroinflammation.Mangiferin,a major glucoside of xanthone in Anemarrhena Rhizome,has anti-inflammatory,anti-diabetes,and anti-oxidative properties.However,the effect of mangiferin on the inflammatary responses of microglia cells are still poorly understand.In this study,we investigated the mechanism by which mangiferin inhibited inflammation in LPS-induced BV2 microglia cells.BV2 cells were pretreatment with mangiferin followed by LPS stimulation.In vitro assays,NO and cytokines production were quantified.Western blot and immunocytochemistry were used to examine the effect of mangiferin on the polarization of BV2 cells and signaling pathway.The results showed that mangiferin treatment significantly reduced NO,IL-1β,IL-6 and TNF-αproduction,also reduced the mRNA and protein of iNOS and COX-2,promoted the polarization of inflammatory toward anti-inflammatory,and inhibited activation of NF-κB and NLRP3 inflammasome.These data suggest that mangiferin has an anti-neuroinflammatory property via regulating microglia macrophage polarization and suppressing NF-κB and NLRP3 signaling pathway,and may act as a potential natural therapeutic candidate for neuroinflammatory diseases.
基金This work was supported by funding from the National Natural Sciences Foundation of China(NSFC 81430021 and 81370470)the Program for Liaoning Innovative Research Team in University(LT2015009)Liaoning Science and Technology Project(2015225008).
文摘Background:Neuron-microglia communication plays a crucial role in the motor neurons(MNs)death in amyotrophic lateral sclerosis(ALS).Neurons can express chemokine(C-X3-C motif)ligand 1(CX3CL1),which mediates microglial activation via interacting with its sole receptor CX3CR1 in microglia.In the present study,we aimed to investigate the dynamic changes of CX3CL1/CX3CR1 axis during microglial activation and MNs loss in SOD1G93A mouse model of ALS.Methods:qPCR,western blot and immunofluorescent staining were used to examine the mRNA and protein levels and localization of CX3CL1/CX3CR1 in the anterior horn region of spinal cord in both SOD1G93A mice and their agematched wild type(WT)littermates at 40,60,90 and 120 days of age.The M1/M2 microglial activation in the spinal cord tissues of SOD1G93A mice and WT mice were evaluated by immunofluorescent staining of M1/M2 markers and further confirmed by qPCR analysis of M1/M2-related cytokines.Results:The immunofluorescent staining revealed that CX3CL1 was predominately expressed in MNs,while CX3CR1 was highly expressed in microglia in the anterior horn region of spinal cord.Compared with age-matched WT mice,CX3CL1 mRNA level was elevated at 40 days but decreased at 90 and 120 days in the anterior horn region of spinal cords in ALS mice.Consistently,CX3CR1 mRNA level was increased at 90 and 120 days.Western blot assay further confirmed the dynamic changes of CX3CL1/CX3CR1 axis in ALS mice.Additionally,the levels of M1/M2 markers of microglia and their related cytokines in the anterior horn region of spinal cord in ALS mice were increased at 90 and 120 days.Moreover,while M1-related cytokines in ALS mice were persistently increased at 120 days,the upregulated M2-related cytokines started to decline at 120 days,suggesting an altered microglial activation.Conclusions:Our data revealed the dynamic changes of CX3CL1/CX3CR1 axis and an imbalanced M1/M2 microglial activation during ALS pathological progression.These findings may help identify potential molecular targets for ALS therapy.
