Satellite glial cells are unique glial cells that surround the cell body of primary sensory neurons.An increasing body of evidence suggests that in the presence of inflammation and nerve damage,a significant number of...Satellite glial cells are unique glial cells that surround the cell body of primary sensory neurons.An increasing body of evidence suggests that in the presence of inflammation and nerve damage,a significant number of satellite glial cells become activated,thus triggering a series of functional changes.This suggests that satellite glial cells are closely related to the occurrence of chronic pain.In this review,we first summarize the morphological structure,molecular markers,and physiological functions of satellite glial cells.Then,we clarify the multiple key roles of satellite glial cells in chronic pain,including gap junction hemichannel Cx43,membrane channel Pannexin1,K channel subunit 4.1,ATP,purinergic P2 receptors,and a series of additional factors and their receptors,including tumor necrosis factor,glutamate,endothelin,and bradykinin.Finally,we propose that future research should focus on the specific sorting of satellite glial cells,and identify genomic differences between physiological and pathological conditions.This review provides an important perspective for clarifying mechanisms underlying the peripheral regulation of chronic pain and will facilitate the formulation of new treatment plans for chronic pain.展开更多
The trigeminal root entry zone is the zone at which the myelination switches from peripheral Schwann cells to central oligodendrocytes.Its special anatomical and physiological structure renders it susceptible to nerve...The trigeminal root entry zone is the zone at which the myelination switches from peripheral Schwann cells to central oligodendrocytes.Its special anatomical and physiological structure renders it susceptible to nerve injury.The etiology of most primary trigeminal neuralgia is closely related to microvascular compression of the trigeminal root entry zone.This study aimed to develop an efficient in vitro model mimicking the glial environment of trigeminal root entry zone as a tool to investigate the effects of glial cell line-derived neurotrophic factor and brain-derived neurotrophic factor on the structural and functional integrity of trigeminal root entry zone and modulation of cellular interactions.Primary astrocytes and Schwann cells isolated from trigeminal root entry zone of postnatal rats were inoculated into a two-well silicon culture insert to mimic the trigeminal root entry zone microenvironment and treated with glial cell line-derived neurotrophic factor and brain-derived neurotrophic factor.In monoculture,glial cell line-derived neurotrophic factor promoted the migration of Schwann cells,but it did not have effects on the migration of astrocytes.In the co-culture system,glial cell line-derived neurotrophic factor promoted the bidirectional migration of astrocytes and Schwann cells.Brain-derived neurotrophic factor markedly promoted the activation and migration of astrocytes.However,in the co-culture system,brain-derived neurotrophic factor inhibited the migration of astrocytes and Schwann cells to a certain degree.These findings suggest that glial cell line-derived neurotrophic factor and brain-derived neurotrophic factor are involved in the regulation of the astrocyte-Schwann cell interaction in the co-culture system derived from the trigeminal root entry zone.This system can be used as a cell model to study the mechanism of glial dysregulation associated with trigeminal nerve injury and possible therapeutic interventions.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
Glial cell line-derived neurotrophic factor recombinant adenovirus vector-transfected bone marrow mesenchymal stem cells were induced to differentiate into neuron-like cells using inductive medium containing retinoic ...Glial cell line-derived neurotrophic factor recombinant adenovirus vector-transfected bone marrow mesenchymal stem cells were induced to differentiate into neuron-like cells using inductive medium containing retinoic acid and epidermal growth factor. Cell viability, micro- tubule-associated protein 2-positive cell ratio, and the expression levels of glial cell line-derived neurotrophic factor, nerve growth factor and growth-associated protein-43 protein in the su- pernatant were significantly higher in glial cell line-derived neurotrophic factor/bone marrow mesenchymal stem cells compared with empty virus plasmid-transfected bone marrow mes- enchymal stem cells. Furthermore, microtubule-associated protein 2, glial cell line-derived neurotrophic factor, nerve growth factor and growth-associated protein743 mRNA levels in cell pellets were statistically higher in glial cell line-derived neurotrophic factor/bone marrow mesen- chymal stem cells compared with empty virus plasmid-transfected bone marrow mesenchymal stem cells. These results suggest that glial cell line-derived neurotrophic factor/bone marrow mesenchymal stem cells have a higher rate of induction into neuron-like cells, and this enhanced differentiation into neuron-like cells may be associated with up-regulated expression of glial cell line-derived neurotrophic factor, nerve growth factor and growth-associated protein-43.展开更多
Mammalian adult central nerve system (CNS) injuries are devastating because of the intrinsic difficulties for effective neuronal regeneration. The greatest problem to be overcome for CNS recovery is the poor regener...Mammalian adult central nerve system (CNS) injuries are devastating because of the intrinsic difficulties for effective neuronal regeneration. The greatest problem to be overcome for CNS recovery is the poor regeneration of neurons and myelin-forming cells, oligodendrocytes. Endogenous neural progenitors and transplanted exogenous neuronal stem cells can be the source for neuronal regeneration. However, because of the harsh local microenvironment, they usually have very low efficacy for functional neural regeneration which cannot compensate for the loss of neurons and oligodendrocytes. Glial cells (including astrocytes, microglia, oligodendrocytes and NG2 glia) are the majority of cells in CNS that provide support and protection for neurons. Inside the local microenvironment, glial cells largely influence local and transplanted neural stem cells survival and fates. This review critically analyzes current finding of the roles of glial cells in CNS regeneration, and highlights strategies for regulating glial cells' behavior to create a permissive microenvironment for neuronal stem cells.展开更多
Summary: Diabetes patients tend to have the gastrointestinal motility disorder. Although the relationship between the motility disorder and both the neurons and Cajal cells in the enteric nervous system (ENS) is we...Summary: Diabetes patients tend to have the gastrointestinal motility disorder. Although the relationship between the motility disorder and both the neurons and Cajal cells in the enteric nervous system (ENS) is well established, little is known about the role of enteric glial cells (EGCs) in gastric motility in diabetes. This study aimed to examine the expression of the glial marker S100B and morphology of EGCs in gastric tissues and the relationship between activated EGCs and the damage of gastric emptying in diabetic models. The diabetic model of rat was induced with 1% streptozotocin (STZ). The model rats at 7-14 days and at 56-63 days were defined as early diabetic rats and advanced diabetic rats, re- spectively, and normal rats at the two time periods served as their corresponding controls. The gastric emptying rate of the rats was tested by using the phenol red solution. The ultrastructure of EGCs in the gastric antrum was observed by the transmission electron microscopy, and the expression of S100B in the myenteric plexus was immunohistochemically detected. The results showed that the gastric emptying rate was significantly increased in the early diabetic rats and decreased in the advanced diabetic rats when compared with their corresponding control rats (P〈0.01 for both). The ultrastructure of EGCs was mostly normal in both the early diabetic and control groups. Vacuolization of mitochondria and expan- sion of endoplasmic reticulum occurred in both the advanced diabetic group and its control group, and even the structure of smooth muscle cells and intestinal neurons was destroyed in the advanced diabetic group. The expression level of S100B in the advanced diabetic group was significantly decreased com- pared with its control group (P〈0.05). It was obviously increased in the early diabetic control group when compared with the advanced diabetic control group (P〈0.05). However, there was no significant difference in the S 100B expression between the early diabetic group and its control group (P〉0.05). The findings suggested that the gastric motility dysfunction in diabetes may be associated with the changes of morphology and number of EGCs in the myenteric plexus.展开更多
To evaluate the effects of glial cell line-derived neurotrophic factor transplantation combined with adipose-derived stem cells-transdifferentiated motoneuron delivery on spinal cord con-tusion injury, we developed ra...To evaluate the effects of glial cell line-derived neurotrophic factor transplantation combined with adipose-derived stem cells-transdifferentiated motoneuron delivery on spinal cord con-tusion injury, we developed rat models of spinal cord contusion injury, 7 days later, injected adipose-derived stem cells-transdifferentiated motoneurons into the epicenter, rostral and caudal regions of the impact site and simultaneously transplanted glial cell line-derived neuro-trophic factor-gelfoam complex into the myelin sheath. Motoneuron-like cell transplantation combined with glial cell line-derived neurotrophic factor delivery reduced cavity formations and increased cell density in the transplantation site. The combined therapy exhibited superior promoting effects on recovery of motor function to transplantation of glial cell line-derived neurotrophic factor, adipose-derived stem cells or motoneurons alone. These ifndings suggest that motoneuron-like cell transplantation combined with glial cell line-derived neurotrophic factor delivery holds a great promise for repair of spinal cord injury.展开更多
BACKGROUND: Substantia nigra is deep in position and limited in range, the glial cell line-derived neurotrophic factor (GDNF) injection directly into substantia nigra has relatively greater damages with higher diff...BACKGROUND: Substantia nigra is deep in position and limited in range, the glial cell line-derived neurotrophic factor (GDNF) injection directly into substantia nigra has relatively greater damages with higher difficulty. GDNF injection into striatum, the target area of dopaminergic neuron, may protect the dopaminergic neurons in the compact part of substantia nigra through retrograde transport. OBJECTIVE: To investigate the protective effect of intrastriatal GDNF on dopaminergic neurons in the substantia nigra of mice with Parkinson disease (PD), and analyze the action pathway. DESIGN: A controlled observation. SETTING: Neurobiological Laboratory of Xuzhou Medical College. MATERIALS: Twenty-four male Kunming mice of 7 - 8 weeks old were used. GDNF, 1-methy1-4-pheny1-1,2,3,6-tetrahydropyridine (MPTP) were purchased from Sigma Company (USA); LEICAQWin image processing and analytical system. METHODS: The experiments were carded out in the Neurobiological Laboratory of Xuzhou Medical College from September 2005 to October 2006. The PD models were established in adult KunMing mice by intraperitoneal injection of MPTP. The model mice were were randomly divided into four groups with 6 mice in each group: GDNF 4-day group, phosphate buffer solution (PSB) 4-day group, GDNF 6-day group and PSB 6-day group. Mice in the GDNF 4 and 6-day groups were administrated with 1 μ L GDNF solution (20 μ g/L, dispensed with 0.01 mol/L PBS) injected into right striatum at 4 and 6 days after model establishment. Mice in the PSB 4 and 6-day groups were administrated with 0.01 mol/L PBS of the same volume to the same injection at corresponding time points. ② On the 12^th day after model establishment, the midbrain tissue section of each mice was divided into 3 areas from rostral to caudal sides. The positive neurons of tyroxine hydroxylase (TH) and calcium binding protein (CB) with obvious nucleolus and clear outline were randomly selected for the measurement, and the number of positive neurons in unit area was counted. MAIN OUTCOME MEASURES: Number of positive neurons of TH and CB in midbrain substantia nigra of mice in each group. RESULTS: All the 24 mice were involved in the analysis of results. The numbers of TH^+ and CB^+ neurons in the GDNF 4-day group (54.33±6.92, 46.33±5.54) were obviously more than those in the PBS 4-day group (27.67±5.01, 21.50±5.96, P 〈 0.01). The numbers of TH^+ and CB^+ neurons in the GDNF 6-day group (75.67±5.39, 69.67±8.69) were obviously more than those in the PBS 6-day group (27.17±4.50, 21.33 ±5.72, P 〈 0.01) and those in the GDNF 4-day group (P 〈 0.01 ). CONCLUSION: Intrastriatal GDNF can protect dopaminergic neurons in substantia nigra of PD mice, and it may be related to the increase of CB expression.展开更多
Hippocampus serves as a pivotal role in cognitive and emotional processes,as well as in the regulation of the hypothalamus-pituitary axis.It is known to undergo mild neurodegenerative changes during normal aging and s...Hippocampus serves as a pivotal role in cognitive and emotional processes,as well as in the regulation of the hypothalamus-pituitary axis.It is known to undergo mild neurodegenerative changes during normal aging and severe atrophy in Alzheimer's disease.Furthermore,dysregulation in the hippocampal function leads to epilepsy and mood disorders.In the first section,we summarized the most salient knowledge on the role of glial cell-line-derived neurotrophic factor and its receptors focused on aging,cognition and neurodegenerative and hippocampal-related neurological diseases mentioned above.In the second section,we reviewed the therapeutic approaches,particularly gene therapy,using glial cell-line-derived neurotrophic factor or its gene,as a key molecule in the development of neurological disorders.In the third section,we pointed at the potential of regenerative medicine,as an emerging and less explored strategy for the treatment of hippocampal disorders.We briefly reviewed the use of partial reprogramming to restore brain functions,non-neuronal cell reprogramming to generate neural stem cells,and neural progenitor cells as source-specific neuronal types to be implanted in animal models of specific neurodegenerative disorders.展开更多
Various retinal injuries induced by ocular hypertension have been shown to induce plastic changes in retinal synapses, but the potential regulatory mechanism of synaptic plasticity after retinal injury was still uncle...Various retinal injuries induced by ocular hypertension have been shown to induce plastic changes in retinal synapses, but the potential regulatory mechanism of synaptic plasticity after retinal injury was still unclear. A rat model of acute ocular hypertension was established by injecting saline intravitreally for an hour, and elevating the intraocular pressure to 14.63 kPa (110 mmHg). Western blot assay and immunofluorescence results showed that synaptophysin expression had a distinct spatiotemporal change that increased in the inner plexiform layer within 1 day and spread across the outer plexiform layer after 3 days. Glial fibrillary acidic protein expression in retinae was greatly increased after 3 days, and reached a peak at 7 days, which was also consistent with the peak time of synaptophysin expression in the outer plexiform layer following the increased intraocular pressure. Fluorocitrate, a glial metabolic inhibitor, was intravitreally injected to inhibit glial cell activation following high intraocular pressure. This significantly inhibited the enhanced glial fibrillary acidic protein expression induced by high intraocular pressure injury. Synaptophysin expression also decreased in the inner plexiform layer within a day and the widened distribution in the outer plexiform layer had disappeared by 3 days. The results suggested that retinal glial cell activation might play an important role in the process of retinal synaptic plasticity induced by acute high intraocular pressure through affecting the expression and distribution of synaptic functional proteins, such as synaptophysin.展开更多
BACKGROUND Slow transit constipation(STC)is a common intestinal disease with increasing incidence.STC results from various factors,such as the enteric nervous system and metabolic changes.As a classical formula of tra...BACKGROUND Slow transit constipation(STC)is a common intestinal disease with increasing incidence.STC results from various factors,such as the enteric nervous system and metabolic changes.As a classical formula of traditional Chinese medicine,Ji-Chuan decoction(JCD)has been extensively and effectively used in STC treatment,yet its pharmacological mechanism remains unclear.AIM To explore the integrated regulatory pattern of JCD against STC through hyphenated techniques from metabolism,network pharmacology and molecular methods.METHODS STC model mice were generated by intragastric administration of compound diphenoxylate(10 mg/kg/d)for 14 d.The STC mice in the low dose of JCD(3.04 g/kg),middle dose of JCD(6.08 g/kg)and high dose of JCD(12.16 g/kg)groups were orally administered JCD solution once a day for 2 wk.The acetylcholine(ACH)level was examined by enzyme-linked immunosorbent assay.The pathological features of colon tissue were observed by hematoxylin and eosin staining.The differentially expressed metabolites and metabolic pathways were tested by nontargeted metabolomics.The main targets and core ingredients of JCD were identified by network pharmacology,and the expression of AKT was confirmed by immunohistochemistry.Finally,the pathways involved in JCD treatment were predicted using a combination of differentially expressed metabolites and targets,and intestinal glial cell apoptosis was demonstrated by immunofluorescence.RESULTS JCD significantly promoted intestinal motility,increased the levels of the excitatory neurotransmitter ACH and reduced intestinal inflammation in STC mice.Untargeted metabolomics results showed that JCD significantly restored metabolic dysfunction and significantly affected taurine and hypotaurine metabolism.Network pharmacology and molecular experiments showed that JCD regulates AKT protein expression,and the core component is quercetin.Combined analysis demonstrated that apoptosis may be an important mechanism by which JCD relieves constipation.Further experiments showed that JCD reduced enteric glial cell(EGC)apoptosis.CONCLUSION This work demonstrated that reducing EGC apoptosis may be the critical mechanism by which JCD treats STC.These findings call for further molecular research to facilitate the clinical application of JCD.展开更多
Spinal cord injury(SCI)is a devastating trauma that currently affects 54 people out of every million,which is approximately 270,000people in the United States(National Spinal Cord Injury Statistical Center,2013).T...Spinal cord injury(SCI)is a devastating trauma that currently affects 54 people out of every million,which is approximately 270,000people in the United States(National Spinal Cord Injury Statistical Center,2013).The effects of such an injury can cause a loss of both motor and sensory function below the injury site,normally leaving the patient unable to care for themselves entirely and relying on family and friends to provide personal care.Currently there are no.展开更多
Several studies have suggested that exogenous glial cell line-derived neurotrophic factor may pro-tect neurons from cerebral ischemic injury. However, the mechanisms underlying the neuroprotec-tive effects of endogeno...Several studies have suggested that exogenous glial cell line-derived neurotrophic factor may pro-tect neurons from cerebral ischemic injury. However, the mechanisms underlying the neuroprotec-tive effects of endogenous glial cell line-derived neurotrophic factor remain unclear. The present experiments sought to elucidate the influence of various conditioned media on neuronal apoptosis, using a normal culture medium for astrocytes, an astrocyte medium highly expressing glial cell line-derived neurotrophic factor, and an astrocyte medium in which glial cell line-derived neurotro-phic factor expression was silenced using RNAi technology. The results confirmed that the use of RNAi silencing to target pretreated glial cell line-derived neurotrophic factor expression promoted neuronal apoptosis. In addition, oxygen and glucose deprivation preconditioning was found to upregulate glial cell line-derived neurotrophic factor expression, and significantly reduce neuronal apoptosis.展开更多
Pitx3 is strongly associated with the phenotype, differentiation, and survival of dopaminergic neurons. The relationship between Pitx3 and glial cell line-derived neurotrophic factor(GDNF) in dopaminergic neurons re...Pitx3 is strongly associated with the phenotype, differentiation, and survival of dopaminergic neurons. The relationship between Pitx3 and glial cell line-derived neurotrophic factor(GDNF) in dopaminergic neurons remains poorly understood. The present investigation sought to construct and screen a lentivirus expression plasmid carrying a rat Pitx3 short hairpin(sh)RNA and to assess the impact of Pitx3 gene knockdown on GDNF transcriptional activity in MES23.5 dopaminergic neurons. Three pairs of interference sequences were designed and separately ligated into GV102 expression vectors. These recombinant plasmids were transfected into MES23.5 cells and western blot assays were performed to detect Pitx3 protein expression. Finally, the most effective Pitx3 sh RNA and a dual-luciferase reporter gene plasmid carrying the GDNF promoter region(GDNF-luciferase) were cotransfected into MES23.5 cells. Sequencing showed that the synthesized sequences were identical to the three Pitx3 interference sequences. Inverted fluorescence microscopy revealed that the lentivirus expression plasmids carrying Pitx3-sh RNA had 40-50% transfection efficiency. Western blot assay confirmed that the corresponding Pitx3 of the third knockdown sequence had the lowest expression level. Dual-luciferase reporter gene results showed that the GDNF transcriptional activity in dopaminergic cells cotransfected with both plasmids was decreased compared with those transfected with GDNF-luciferase alone. Together, the results showed that the designed Pitx3-sh RNA interference sequence decreased Pitx3 protein expression, which decreased GDNF transcriptional activity.展开更多
The present study co-cultured human embryonic olfactory ensheathJng cells, human Schwann cells, human amniotic epithelial cells and human vascular endothelial cells in complete culture medium- containing cerebrospinal...The present study co-cultured human embryonic olfactory ensheathJng cells, human Schwann cells, human amniotic epithelial cells and human vascular endothelial cells in complete culture medium- containing cerebrospinal fluid. Enzyme linked immunosorbent assay was used to detect nerve growth factor, brain-derived neurotrophic factor, and glial cell line-derived neurotrophic factor secretion in the supernatant of co-cultured cells. Results showed that the number of all cell types reached a peak at 7-10 days, and the expression of nerve growth factor, brain-derived neurotrophic factor, and glial cell line-derived neurotrophic factor peaked at 9 days. Levels of secreted nerve growth factor were four-fold higher than brain-derived neurotrophic factor, which was three-fold higher than glial cell line-derived neurotrophic factor. Increasing concentrations of cerebrospinal fluid (10%, 20% and 30%) in the growth medium caused a decrease of neurotrophic factor secretion Results indicated co-culture of human embryonic olfactory ensheathing cells, human Schwann cells human amniotic epithelial cells and human vascular endothelial cells improved the expression of nerve growth factor, brain-derived neurotrophic factor, and glial cell line-derived neurotrophic factor. The reduction of cerebrospinal fluid extravasation at the transplant site after spinal cord injury is beneficial for the survival and secretion of neurotrophic factors from transplanted cells.展开更多
The following mini-review attempts to guide researchers in the quantification of fluorescently-labelled proteins within cultured thick or chromogenically-stained proteins within thin sections of brain tissue.It follow...The following mini-review attempts to guide researchers in the quantification of fluorescently-labelled proteins within cultured thick or chromogenically-stained proteins within thin sections of brain tissue.It follows from our examination of the utility of Fiji Image J thresholding and binarization algorithms.Describing how we identified the maximum intensity projection as the best of six tested for two dimensional(2 D)-rendering of three-dimensional(3 D) images derived from a series of z-stacked micrographs,the review summarises our comparison of 16 global and 9 local algorithms for their ability to accurately quantify the expression of astrocytic glial fibrillary acidic protein(GFAP),microglial ionized calcium binding adapter molecule 1(IBA1) and oligodendrocyte lineage Olig2 within fixed cultured rat hippocampal brain slices.The application of these algorithms to chromogenically-stained GFAP and IBA1 within thin tissue sections,is also described.Fiji’s Bio Voxxel plugin allowed categorisation of algorithms according to their sensitivity,specificity accuracy and relative quality.The Percentile algorithm was deemed best for quantifying levels of GFAP,the Li algorithm was best when quantifying IBA expression,while the Otsu algorithm was optimum for Olig2 staining,albeit with over-quantification of oligodendrocyte number when compared to a stereological approach.Also,GFAP and IBA expression in 3,3′-diaminobenzidine(DAB)/haematoxylin-stained cerebellar tissue was best quantified with Default,Isodata and Moments algorithms.The workflow presented in Figure 1 could help to improve the quality of research outcomes that are based on the quantification of protein with brain tissue.展开更多
Objective:To evaluate whether ginsenoside Rb1(Rb1) can attenuate lipopolysaccharide(LPS)-induced chronic neuroinflammation in mice and to explore its relationship with glial cell polarization.Methods:Intraperitoneal i...Objective:To evaluate whether ginsenoside Rb1(Rb1) can attenuate lipopolysaccharide(LPS)-induced chronic neuroinflammation in mice and to explore its relationship with glial cell polarization.Methods:Intraperitoneal injection with an escalating dose of LPS was used to establish a chronic neuroinflammation model in mice.Once LPS was initiated,10 or 20 mg/kg Rbl,or sterile saline,was administered for 14 consecutive days.Open field test and beam walking test were used to monitor the changes in behavior.The concentration of cytokines in the serum and brain were used to monitor the systemic inflammation and neuroinflammation,respectively.Molecules specific to each glial cell phenotype were used to investigate glial cell polarization.Results:Mice in the LPS group had reduced spontaneous activities and impaired beam walking performance.Rbl obviously eased LPS-induced behavior distu rbances.Regarding the levels of serum cytokines,both tumor necrosis factor-α(TNF-α) and interleukin-1β(IL-1β) were significantly increased,while interleukin-10(IL-10) and transforming growth factor β(TGF-β) remarkably decreased after LPS treatment(all P <.001).Rb1 treatment significantly attenuated LPS-induced serum cytokine changes(all P <.05).The results of quantitative polymerase chain reaction and western blotting showed that the mRNA and protein expression levels of TNF-α and complement component 3(C3) in the brain were significantly increased after LPS treatment(all P<.01).Rbl treatment significantly inhibited LPS-induced inflammation in the brain(all P <.05).Glial cell polarization analysis showed that M1 and M2 microglia,and A1 astrocytes increased following LPS treatment,while A2 astrocytes decreased.Rb1 treatment reduced M1 and M2 microglia,and A1 astrocytes,and significantly increased A2 astrocytes.Conclusion:Rb1 can attenuate chronic neuroinflammation induced by LPS in mice,which may be partially attributable to its fine tuning of microglia and astrocyte polarization.Rb1 has potential value for treating neurodegenerative diseases.展开更多
BACKGROUND Parkinson’s disease(PD)is a neurological disorder characterized by the progressive loss of midbrain dopamine(DA)neurons.Bone marrow mesenchymal stem cells(BMSCs)can differentiate into multiple cell types i...BACKGROUND Parkinson’s disease(PD)is a neurological disorder characterized by the progressive loss of midbrain dopamine(DA)neurons.Bone marrow mesenchymal stem cells(BMSCs)can differentiate into multiple cell types including neurons and glia.Transplantation of BMSCs is regarded as a potential approach for promoting neural regeneration.Glial cell line-derived neurotrophic factor(GDNF)can induce BMSC differentiation into neuron-like cells.This work evaluated the efficacy of nigral grafts of human BMSCs(hMSCs)and/or adenoviral(Ad)GDNF gene transfer in 6-hydroxydopamine(6-OHDA)-lesioned hemiparkinsonian rats.AIM To evaluate the efficacy of nigral grafts of hMSCs and/or Ad-GDNF gene transfer in 6-OHDA-lesioned hemiparkinsonian rats.METHODS We used immortalized hMSCs,which retain their potential for neuronal differentiation.hMSCs,preinduced hMSCs,or Ad-GDNF effectively enhanced neuronal connections in cultured neurons.In vivo,preinduced hMSCs and/or Ad-GDNF were injected into the substantia nigra(SN)after induction of a unilateral 6-OHDA lesion in the nigrostriatal pathway.RESULTS Hemiparkinsonian rats that received preinduced hMSC graft and/or Ad-GDNF showed significant recovery of apomorphine-induced rotational behavior and the number of nigral DA neurons.However,DA levels in the striatum were not restored by these therapeutic treatments.Grafted hMSCs might reconstitute a niche to support tissue repair rather than contribute to the generation of new neurons in the injured SN.CONCLUSION The results suggest that preinduced hMSC grafts exert a regenerative effect and may have the potential to improve clinical outcome.展开更多
Multiple sclerosis (MS) is a chronic inflammatory and neurodegenerative disorder that is thought to be mediated by autoreactive T lymphocytes that find their way into the central nervous system (CNS). The patholog...Multiple sclerosis (MS) is a chronic inflammatory and neurodegenerative disorder that is thought to be mediated by autoreactive T lymphocytes that find their way into the central nervous system (CNS). The pathological mechanism of MS is still being elucidated but it involves complex interactions between infiltrating immune cells and resi- dent glial cells within the CNS that culminate into strong neuroinflammation and axonal damage.展开更多
基金supported by the National Natural Science Foundation of China,No.82173796(to CX)the Research Project of Zhejiang Chinese Medical University,No.2022JKJNTZ13(to XQ).
文摘Satellite glial cells are unique glial cells that surround the cell body of primary sensory neurons.An increasing body of evidence suggests that in the presence of inflammation and nerve damage,a significant number of satellite glial cells become activated,thus triggering a series of functional changes.This suggests that satellite glial cells are closely related to the occurrence of chronic pain.In this review,we first summarize the morphological structure,molecular markers,and physiological functions of satellite glial cells.Then,we clarify the multiple key roles of satellite glial cells in chronic pain,including gap junction hemichannel Cx43,membrane channel Pannexin1,K channel subunit 4.1,ATP,purinergic P2 receptors,and a series of additional factors and their receptors,including tumor necrosis factor,glutamate,endothelin,and bradykinin.Finally,we propose that future research should focus on the specific sorting of satellite glial cells,and identify genomic differences between physiological and pathological conditions.This review provides an important perspective for clarifying mechanisms underlying the peripheral regulation of chronic pain and will facilitate the formulation of new treatment plans for chronic pain.
基金supported by the National Natural Sclence Foundation of China in 2021No.82171213+1 种基金the Natural Science Foundation of Fujian Province in 2019No.2019J01289 (both to DSL)
文摘The trigeminal root entry zone is the zone at which the myelination switches from peripheral Schwann cells to central oligodendrocytes.Its special anatomical and physiological structure renders it susceptible to nerve injury.The etiology of most primary trigeminal neuralgia is closely related to microvascular compression of the trigeminal root entry zone.This study aimed to develop an efficient in vitro model mimicking the glial environment of trigeminal root entry zone as a tool to investigate the effects of glial cell line-derived neurotrophic factor and brain-derived neurotrophic factor on the structural and functional integrity of trigeminal root entry zone and modulation of cellular interactions.Primary astrocytes and Schwann cells isolated from trigeminal root entry zone of postnatal rats were inoculated into a two-well silicon culture insert to mimic the trigeminal root entry zone microenvironment and treated with glial cell line-derived neurotrophic factor and brain-derived neurotrophic factor.In monoculture,glial cell line-derived neurotrophic factor promoted the migration of Schwann cells,but it did not have effects on the migration of astrocytes.In the co-culture system,glial cell line-derived neurotrophic factor promoted the bidirectional migration of astrocytes and Schwann cells.Brain-derived neurotrophic factor markedly promoted the activation and migration of astrocytes.However,in the co-culture system,brain-derived neurotrophic factor inhibited the migration of astrocytes and Schwann cells to a certain degree.These findings suggest that glial cell line-derived neurotrophic factor and brain-derived neurotrophic factor are involved in the regulation of the astrocyte-Schwann cell interaction in the co-culture system derived from the trigeminal root entry zone.This system can be used as a cell model to study the mechanism of glial dysregulation associated with trigeminal nerve injury and possible therapeutic interventions.
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.
文摘Glial cell line-derived neurotrophic factor recombinant adenovirus vector-transfected bone marrow mesenchymal stem cells were induced to differentiate into neuron-like cells using inductive medium containing retinoic acid and epidermal growth factor. Cell viability, micro- tubule-associated protein 2-positive cell ratio, and the expression levels of glial cell line-derived neurotrophic factor, nerve growth factor and growth-associated protein-43 protein in the su- pernatant were significantly higher in glial cell line-derived neurotrophic factor/bone marrow mesenchymal stem cells compared with empty virus plasmid-transfected bone marrow mes- enchymal stem cells. Furthermore, microtubule-associated protein 2, glial cell line-derived neurotrophic factor, nerve growth factor and growth-associated protein743 mRNA levels in cell pellets were statistically higher in glial cell line-derived neurotrophic factor/bone marrow mesen- chymal stem cells compared with empty virus plasmid-transfected bone marrow mesenchymal stem cells. These results suggest that glial cell line-derived neurotrophic factor/bone marrow mesenchymal stem cells have a higher rate of induction into neuron-like cells, and this enhanced differentiation into neuron-like cells may be associated with up-regulated expression of glial cell line-derived neurotrophic factor, nerve growth factor and growth-associated protein-43.
基金supported in part by the Nippon Dental University Research Project 4 Grant and Japan Society for the Promotion of Science(JSPS)Grant-in-Aid for Scientific Research(26861689)
文摘Mammalian adult central nerve system (CNS) injuries are devastating because of the intrinsic difficulties for effective neuronal regeneration. The greatest problem to be overcome for CNS recovery is the poor regeneration of neurons and myelin-forming cells, oligodendrocytes. Endogenous neural progenitors and transplanted exogenous neuronal stem cells can be the source for neuronal regeneration. However, because of the harsh local microenvironment, they usually have very low efficacy for functional neural regeneration which cannot compensate for the loss of neurons and oligodendrocytes. Glial cells (including astrocytes, microglia, oligodendrocytes and NG2 glia) are the majority of cells in CNS that provide support and protection for neurons. Inside the local microenvironment, glial cells largely influence local and transplanted neural stem cells survival and fates. This review critically analyzes current finding of the roles of glial cells in CNS regeneration, and highlights strategies for regulating glial cells' behavior to create a permissive microenvironment for neuronal stem cells.
基金supported by a grant from the National Nature Science Foundation of China(No.81170342)
文摘Summary: Diabetes patients tend to have the gastrointestinal motility disorder. Although the relationship between the motility disorder and both the neurons and Cajal cells in the enteric nervous system (ENS) is well established, little is known about the role of enteric glial cells (EGCs) in gastric motility in diabetes. This study aimed to examine the expression of the glial marker S100B and morphology of EGCs in gastric tissues and the relationship between activated EGCs and the damage of gastric emptying in diabetic models. The diabetic model of rat was induced with 1% streptozotocin (STZ). The model rats at 7-14 days and at 56-63 days were defined as early diabetic rats and advanced diabetic rats, re- spectively, and normal rats at the two time periods served as their corresponding controls. The gastric emptying rate of the rats was tested by using the phenol red solution. The ultrastructure of EGCs in the gastric antrum was observed by the transmission electron microscopy, and the expression of S100B in the myenteric plexus was immunohistochemically detected. The results showed that the gastric emptying rate was significantly increased in the early diabetic rats and decreased in the advanced diabetic rats when compared with their corresponding control rats (P〈0.01 for both). The ultrastructure of EGCs was mostly normal in both the early diabetic and control groups. Vacuolization of mitochondria and expan- sion of endoplasmic reticulum occurred in both the advanced diabetic group and its control group, and even the structure of smooth muscle cells and intestinal neurons was destroyed in the advanced diabetic group. The expression level of S100B in the advanced diabetic group was significantly decreased com- pared with its control group (P〈0.05). It was obviously increased in the early diabetic control group when compared with the advanced diabetic control group (P〈0.05). However, there was no significant difference in the S 100B expression between the early diabetic group and its control group (P〉0.05). The findings suggested that the gastric motility dysfunction in diabetes may be associated with the changes of morphology and number of EGCs in the myenteric plexus.
基金funded by Shefa Neurosciences Research Center at Khatam Al-Anbia Hospital,Tehran,Iran(Grant#86-N-105)
文摘To evaluate the effects of glial cell line-derived neurotrophic factor transplantation combined with adipose-derived stem cells-transdifferentiated motoneuron delivery on spinal cord con-tusion injury, we developed rat models of spinal cord contusion injury, 7 days later, injected adipose-derived stem cells-transdifferentiated motoneurons into the epicenter, rostral and caudal regions of the impact site and simultaneously transplanted glial cell line-derived neuro-trophic factor-gelfoam complex into the myelin sheath. Motoneuron-like cell transplantation combined with glial cell line-derived neurotrophic factor delivery reduced cavity formations and increased cell density in the transplantation site. The combined therapy exhibited superior promoting effects on recovery of motor function to transplantation of glial cell line-derived neurotrophic factor, adipose-derived stem cells or motoneurons alone. These ifndings suggest that motoneuron-like cell transplantation combined with glial cell line-derived neurotrophic factor delivery holds a great promise for repair of spinal cord injury.
基金the Natural Science Foundation of Jiangsu Department of Education, No. 02KJB310009
文摘BACKGROUND: Substantia nigra is deep in position and limited in range, the glial cell line-derived neurotrophic factor (GDNF) injection directly into substantia nigra has relatively greater damages with higher difficulty. GDNF injection into striatum, the target area of dopaminergic neuron, may protect the dopaminergic neurons in the compact part of substantia nigra through retrograde transport. OBJECTIVE: To investigate the protective effect of intrastriatal GDNF on dopaminergic neurons in the substantia nigra of mice with Parkinson disease (PD), and analyze the action pathway. DESIGN: A controlled observation. SETTING: Neurobiological Laboratory of Xuzhou Medical College. MATERIALS: Twenty-four male Kunming mice of 7 - 8 weeks old were used. GDNF, 1-methy1-4-pheny1-1,2,3,6-tetrahydropyridine (MPTP) were purchased from Sigma Company (USA); LEICAQWin image processing and analytical system. METHODS: The experiments were carded out in the Neurobiological Laboratory of Xuzhou Medical College from September 2005 to October 2006. The PD models were established in adult KunMing mice by intraperitoneal injection of MPTP. The model mice were were randomly divided into four groups with 6 mice in each group: GDNF 4-day group, phosphate buffer solution (PSB) 4-day group, GDNF 6-day group and PSB 6-day group. Mice in the GDNF 4 and 6-day groups were administrated with 1 μ L GDNF solution (20 μ g/L, dispensed with 0.01 mol/L PBS) injected into right striatum at 4 and 6 days after model establishment. Mice in the PSB 4 and 6-day groups were administrated with 0.01 mol/L PBS of the same volume to the same injection at corresponding time points. ② On the 12^th day after model establishment, the midbrain tissue section of each mice was divided into 3 areas from rostral to caudal sides. The positive neurons of tyroxine hydroxylase (TH) and calcium binding protein (CB) with obvious nucleolus and clear outline were randomly selected for the measurement, and the number of positive neurons in unit area was counted. MAIN OUTCOME MEASURES: Number of positive neurons of TH and CB in midbrain substantia nigra of mice in each group. RESULTS: All the 24 mice were involved in the analysis of results. The numbers of TH^+ and CB^+ neurons in the GDNF 4-day group (54.33±6.92, 46.33±5.54) were obviously more than those in the PBS 4-day group (27.67±5.01, 21.50±5.96, P 〈 0.01). The numbers of TH^+ and CB^+ neurons in the GDNF 6-day group (75.67±5.39, 69.67±8.69) were obviously more than those in the PBS 6-day group (27.17±4.50, 21.33 ±5.72, P 〈 0.01) and those in the GDNF 4-day group (P 〈 0.01 ). CONCLUSION: Intrastriatal GDNF can protect dopaminergic neurons in substantia nigra of PD mice, and it may be related to the increase of CB expression.
基金supported by grant #PICT16-1070 from the Argentine Agency for the Promotion of Science and Technology (ANCYPT) to GRM。
文摘Hippocampus serves as a pivotal role in cognitive and emotional processes,as well as in the regulation of the hypothalamus-pituitary axis.It is known to undergo mild neurodegenerative changes during normal aging and severe atrophy in Alzheimer's disease.Furthermore,dysregulation in the hippocampal function leads to epilepsy and mood disorders.In the first section,we summarized the most salient knowledge on the role of glial cell-line-derived neurotrophic factor and its receptors focused on aging,cognition and neurodegenerative and hippocampal-related neurological diseases mentioned above.In the second section,we reviewed the therapeutic approaches,particularly gene therapy,using glial cell-line-derived neurotrophic factor or its gene,as a key molecule in the development of neurological disorders.In the third section,we pointed at the potential of regenerative medicine,as an emerging and less explored strategy for the treatment of hippocampal disorders.We briefly reviewed the use of partial reprogramming to restore brain functions,non-neuronal cell reprogramming to generate neural stem cells,and neural progenitor cells as source-specific neuronal types to be implanted in animal models of specific neurodegenerative disorders.
基金supported by the National Natural Science Foundation of China,No.81070729the Natural Science Foundation of Hunan Province in China,No.10JJ4023the Hunan Provincial Innovation Foundation for Postgraduate in China,No.CX2011B047
文摘Various retinal injuries induced by ocular hypertension have been shown to induce plastic changes in retinal synapses, but the potential regulatory mechanism of synaptic plasticity after retinal injury was still unclear. A rat model of acute ocular hypertension was established by injecting saline intravitreally for an hour, and elevating the intraocular pressure to 14.63 kPa (110 mmHg). Western blot assay and immunofluorescence results showed that synaptophysin expression had a distinct spatiotemporal change that increased in the inner plexiform layer within 1 day and spread across the outer plexiform layer after 3 days. Glial fibrillary acidic protein expression in retinae was greatly increased after 3 days, and reached a peak at 7 days, which was also consistent with the peak time of synaptophysin expression in the outer plexiform layer following the increased intraocular pressure. Fluorocitrate, a glial metabolic inhibitor, was intravitreally injected to inhibit glial cell activation following high intraocular pressure. This significantly inhibited the enhanced glial fibrillary acidic protein expression induced by high intraocular pressure injury. Synaptophysin expression also decreased in the inner plexiform layer within a day and the widened distribution in the outer plexiform layer had disappeared by 3 days. The results suggested that retinal glial cell activation might play an important role in the process of retinal synaptic plasticity induced by acute high intraocular pressure through affecting the expression and distribution of synaptic functional proteins, such as synaptophysin.
基金Supported by the National Natural Science Foundation of China,No. 82074151the Experimental Formulary Sichuan Youth Science and Technology Innovation Research Team,No. 2020JDTD0022
文摘BACKGROUND Slow transit constipation(STC)is a common intestinal disease with increasing incidence.STC results from various factors,such as the enteric nervous system and metabolic changes.As a classical formula of traditional Chinese medicine,Ji-Chuan decoction(JCD)has been extensively and effectively used in STC treatment,yet its pharmacological mechanism remains unclear.AIM To explore the integrated regulatory pattern of JCD against STC through hyphenated techniques from metabolism,network pharmacology and molecular methods.METHODS STC model mice were generated by intragastric administration of compound diphenoxylate(10 mg/kg/d)for 14 d.The STC mice in the low dose of JCD(3.04 g/kg),middle dose of JCD(6.08 g/kg)and high dose of JCD(12.16 g/kg)groups were orally administered JCD solution once a day for 2 wk.The acetylcholine(ACH)level was examined by enzyme-linked immunosorbent assay.The pathological features of colon tissue were observed by hematoxylin and eosin staining.The differentially expressed metabolites and metabolic pathways were tested by nontargeted metabolomics.The main targets and core ingredients of JCD were identified by network pharmacology,and the expression of AKT was confirmed by immunohistochemistry.Finally,the pathways involved in JCD treatment were predicted using a combination of differentially expressed metabolites and targets,and intestinal glial cell apoptosis was demonstrated by immunofluorescence.RESULTS JCD significantly promoted intestinal motility,increased the levels of the excitatory neurotransmitter ACH and reduced intestinal inflammation in STC mice.Untargeted metabolomics results showed that JCD significantly restored metabolic dysfunction and significantly affected taurine and hypotaurine metabolism.Network pharmacology and molecular experiments showed that JCD regulates AKT protein expression,and the core component is quercetin.Combined analysis demonstrated that apoptosis may be an important mechanism by which JCD relieves constipation.Further experiments showed that JCD reduced enteric glial cell(EGC)apoptosis.CONCLUSION This work demonstrated that reducing EGC apoptosis may be the critical mechanism by which JCD treats STC.These findings call for further molecular research to facilitate the clinical application of JCD.
文摘Spinal cord injury(SCI)is a devastating trauma that currently affects 54 people out of every million,which is approximately 270,000people in the United States(National Spinal Cord Injury Statistical Center,2013).The effects of such an injury can cause a loss of both motor and sensory function below the injury site,normally leaving the patient unable to care for themselves entirely and relying on family and friends to provide personal care.Currently there are no.
基金Specialized Research Fund for the Doc-toral Program of Higher Education, No. 20060183053
文摘Several studies have suggested that exogenous glial cell line-derived neurotrophic factor may pro-tect neurons from cerebral ischemic injury. However, the mechanisms underlying the neuroprotec-tive effects of endogenous glial cell line-derived neurotrophic factor remain unclear. The present experiments sought to elucidate the influence of various conditioned media on neuronal apoptosis, using a normal culture medium for astrocytes, an astrocyte medium highly expressing glial cell line-derived neurotrophic factor, and an astrocyte medium in which glial cell line-derived neurotro-phic factor expression was silenced using RNAi technology. The results confirmed that the use of RNAi silencing to target pretreated glial cell line-derived neurotrophic factor expression promoted neuronal apoptosis. In addition, oxygen and glucose deprivation preconditioning was found to upregulate glial cell line-derived neurotrophic factor expression, and significantly reduce neuronal apoptosis.
基金supported by the National Natural Science Foundation of China,No.81372698
文摘Pitx3 is strongly associated with the phenotype, differentiation, and survival of dopaminergic neurons. The relationship between Pitx3 and glial cell line-derived neurotrophic factor(GDNF) in dopaminergic neurons remains poorly understood. The present investigation sought to construct and screen a lentivirus expression plasmid carrying a rat Pitx3 short hairpin(sh)RNA and to assess the impact of Pitx3 gene knockdown on GDNF transcriptional activity in MES23.5 dopaminergic neurons. Three pairs of interference sequences were designed and separately ligated into GV102 expression vectors. These recombinant plasmids were transfected into MES23.5 cells and western blot assays were performed to detect Pitx3 protein expression. Finally, the most effective Pitx3 sh RNA and a dual-luciferase reporter gene plasmid carrying the GDNF promoter region(GDNF-luciferase) were cotransfected into MES23.5 cells. Sequencing showed that the synthesized sequences were identical to the three Pitx3 interference sequences. Inverted fluorescence microscopy revealed that the lentivirus expression plasmids carrying Pitx3-sh RNA had 40-50% transfection efficiency. Western blot assay confirmed that the corresponding Pitx3 of the third knockdown sequence had the lowest expression level. Dual-luciferase reporter gene results showed that the GDNF transcriptional activity in dopaminergic cells cotransfected with both plasmids was decreased compared with those transfected with GDNF-luciferase alone. Together, the results showed that the designed Pitx3-sh RNA interference sequence decreased Pitx3 protein expression, which decreased GDNF transcriptional activity.
基金supported by the Science andTechnology Development Program of Guangdong Province, No.2009b030801329
文摘The present study co-cultured human embryonic olfactory ensheathJng cells, human Schwann cells, human amniotic epithelial cells and human vascular endothelial cells in complete culture medium- containing cerebrospinal fluid. Enzyme linked immunosorbent assay was used to detect nerve growth factor, brain-derived neurotrophic factor, and glial cell line-derived neurotrophic factor secretion in the supernatant of co-cultured cells. Results showed that the number of all cell types reached a peak at 7-10 days, and the expression of nerve growth factor, brain-derived neurotrophic factor, and glial cell line-derived neurotrophic factor peaked at 9 days. Levels of secreted nerve growth factor were four-fold higher than brain-derived neurotrophic factor, which was three-fold higher than glial cell line-derived neurotrophic factor. Increasing concentrations of cerebrospinal fluid (10%, 20% and 30%) in the growth medium caused a decrease of neurotrophic factor secretion Results indicated co-culture of human embryonic olfactory ensheathing cells, human Schwann cells human amniotic epithelial cells and human vascular endothelial cells improved the expression of nerve growth factor, brain-derived neurotrophic factor, and glial cell line-derived neurotrophic factor. The reduction of cerebrospinal fluid extravasation at the transplant site after spinal cord injury is beneficial for the survival and secretion of neurotrophic factors from transplanted cells.
基金supported by a grant from Thomas Crawford Hayes Research Fundthe NUI Galway College of Science scholarship to SHa grant from NUI Galway Foundation Office to JM
文摘The following mini-review attempts to guide researchers in the quantification of fluorescently-labelled proteins within cultured thick or chromogenically-stained proteins within thin sections of brain tissue.It follows from our examination of the utility of Fiji Image J thresholding and binarization algorithms.Describing how we identified the maximum intensity projection as the best of six tested for two dimensional(2 D)-rendering of three-dimensional(3 D) images derived from a series of z-stacked micrographs,the review summarises our comparison of 16 global and 9 local algorithms for their ability to accurately quantify the expression of astrocytic glial fibrillary acidic protein(GFAP),microglial ionized calcium binding adapter molecule 1(IBA1) and oligodendrocyte lineage Olig2 within fixed cultured rat hippocampal brain slices.The application of these algorithms to chromogenically-stained GFAP and IBA1 within thin tissue sections,is also described.Fiji’s Bio Voxxel plugin allowed categorisation of algorithms according to their sensitivity,specificity accuracy and relative quality.The Percentile algorithm was deemed best for quantifying levels of GFAP,the Li algorithm was best when quantifying IBA expression,while the Otsu algorithm was optimum for Olig2 staining,albeit with over-quantification of oligodendrocyte number when compared to a stereological approach.Also,GFAP and IBA expression in 3,3′-diaminobenzidine(DAB)/haematoxylin-stained cerebellar tissue was best quantified with Default,Isodata and Moments algorithms.The workflow presented in Figure 1 could help to improve the quality of research outcomes that are based on the quantification of protein with brain tissue.
基金This research was supported by the grant“Prevention and Treatment of Major Diseases with Chinese Medicine from the National Administration of Traditional Chinese Medicine(GZYYGJ2021047)”the“High-end Experts Support Program from the Ministry of Science and Technology(DL 2021110001L)”the“Basic Research Funds from the Ministry of Education 1000061223731)”。
文摘Objective:To evaluate whether ginsenoside Rb1(Rb1) can attenuate lipopolysaccharide(LPS)-induced chronic neuroinflammation in mice and to explore its relationship with glial cell polarization.Methods:Intraperitoneal injection with an escalating dose of LPS was used to establish a chronic neuroinflammation model in mice.Once LPS was initiated,10 or 20 mg/kg Rbl,or sterile saline,was administered for 14 consecutive days.Open field test and beam walking test were used to monitor the changes in behavior.The concentration of cytokines in the serum and brain were used to monitor the systemic inflammation and neuroinflammation,respectively.Molecules specific to each glial cell phenotype were used to investigate glial cell polarization.Results:Mice in the LPS group had reduced spontaneous activities and impaired beam walking performance.Rbl obviously eased LPS-induced behavior distu rbances.Regarding the levels of serum cytokines,both tumor necrosis factor-α(TNF-α) and interleukin-1β(IL-1β) were significantly increased,while interleukin-10(IL-10) and transforming growth factor β(TGF-β) remarkably decreased after LPS treatment(all P <.001).Rb1 treatment significantly attenuated LPS-induced serum cytokine changes(all P <.05).The results of quantitative polymerase chain reaction and western blotting showed that the mRNA and protein expression levels of TNF-α and complement component 3(C3) in the brain were significantly increased after LPS treatment(all P<.01).Rbl treatment significantly inhibited LPS-induced inflammation in the brain(all P <.05).Glial cell polarization analysis showed that M1 and M2 microglia,and A1 astrocytes increased following LPS treatment,while A2 astrocytes decreased.Rb1 treatment reduced M1 and M2 microglia,and A1 astrocytes,and significantly increased A2 astrocytes.Conclusion:Rb1 can attenuate chronic neuroinflammation induced by LPS in mice,which may be partially attributable to its fine tuning of microglia and astrocyte polarization.Rb1 has potential value for treating neurodegenerative diseases.
基金Supported by Taipei Veterans General Hospital in Taiwan,No.V106C-012,No.V107C-087,and No.V109C-018and Ministry of Science and Technology in Taiwan,No.MOST106-2314-B-075-023,No.MOST107-2314-B-010-023,and No.MOST107-2314-B-075-021.
文摘BACKGROUND Parkinson’s disease(PD)is a neurological disorder characterized by the progressive loss of midbrain dopamine(DA)neurons.Bone marrow mesenchymal stem cells(BMSCs)can differentiate into multiple cell types including neurons and glia.Transplantation of BMSCs is regarded as a potential approach for promoting neural regeneration.Glial cell line-derived neurotrophic factor(GDNF)can induce BMSC differentiation into neuron-like cells.This work evaluated the efficacy of nigral grafts of human BMSCs(hMSCs)and/or adenoviral(Ad)GDNF gene transfer in 6-hydroxydopamine(6-OHDA)-lesioned hemiparkinsonian rats.AIM To evaluate the efficacy of nigral grafts of hMSCs and/or Ad-GDNF gene transfer in 6-OHDA-lesioned hemiparkinsonian rats.METHODS We used immortalized hMSCs,which retain their potential for neuronal differentiation.hMSCs,preinduced hMSCs,or Ad-GDNF effectively enhanced neuronal connections in cultured neurons.In vivo,preinduced hMSCs and/or Ad-GDNF were injected into the substantia nigra(SN)after induction of a unilateral 6-OHDA lesion in the nigrostriatal pathway.RESULTS Hemiparkinsonian rats that received preinduced hMSC graft and/or Ad-GDNF showed significant recovery of apomorphine-induced rotational behavior and the number of nigral DA neurons.However,DA levels in the striatum were not restored by these therapeutic treatments.Grafted hMSCs might reconstitute a niche to support tissue repair rather than contribute to the generation of new neurons in the injured SN.CONCLUSION The results suggest that preinduced hMSC grafts exert a regenerative effect and may have the potential to improve clinical outcome.
基金supported by the Helmholtz-Gemeinschaft,“Zukunft-sthema”Immunology and inflammation”(ZT-0027)supported by the Pertermax-Müller-Stiftung and the Niedersachsen Research Network on Neuroinfectiology(N-RENNT)of the Ministry of Science and Culture of Lower Saxony
文摘Multiple sclerosis (MS) is a chronic inflammatory and neurodegenerative disorder that is thought to be mediated by autoreactive T lymphocytes that find their way into the central nervous system (CNS). The pathological mechanism of MS is still being elucidated but it involves complex interactions between infiltrating immune cells and resi- dent glial cells within the CNS that culminate into strong neuroinflammation and axonal damage.