Single-cell profiling reveals Müller glia coordinate retinal intercellular communication during light/dark adaptation via thyroid hormone signaling

Light adaptation enables the vertebrate visual system to operate over a wide range of ambient illumination. Regulation of phototransduction in photoreceptors is considered a major mechanism underlying light adaptation. However, various types of neurons and glial cells exist in the retina, and whether and how all retinal cells interact to adapt to light/dark conditions at the cellular and molecular levels requires systematic investigation. Therefore, we utilized single-cell RNA sequencing to dissect retinal cell-type-specific transcriptomes during light/dark adaptation in mice. The results demonstrated that, in addition to photoreceptors, other retinal cell types also showed dynamic molecular changes and specifically enriched signaling pathways under light/dark adaptation. Importantly, Müller glial cells (MGs) were identified as hub cells for intercellular interactions, displaying complex cell‒cell communication with other retinal cells. Furthermore, light increased the transcription of the deiodinase Dio2 in MGs, which converted thyroxine (T4) to active triiodothyronine (T3). Subsequently, light increased T3 levels and regulated mitochondrial respiration in retinal cells in response to light conditions. As cones specifically express the thyroid hormone receptor Thrb, they responded to the increase in T3 by adjusting light responsiveness. Loss of the expression of Dio2 specifically in MGs decreased the light responsive ability of cones. These results suggest that retinal cells display global transcriptional changes under light/dark adaptation and that MGs coordinate intercellular communication during light/dark adaptation via thyroid hormone signaling.

Research progress on the nerve damage in diabetic retinopathy / 国际眼科杂志(Guoji Yanke Zazhi)

Diabetic retinopathy(DR)is the main cause of substantial visual impairment of occupational active individuals in the world, which has become one of the most common eye diseases that lead to irreversible visual impairment of the working population. Precise identification and accurate intervention of early DR lesions are of great significance to block or delay the development of this disease. Recent studies have shown that DR nerve injury occurs before retinal microangiopathy, it has a series of characteristic clinical manifestations, such as dark adaptation delay, contrast sensitivity and decreased tone discrimination. These characteristic clinical manifestations are key events in the early stage of DR, which are closely related to apoptosis, glial cell proliferation, oxidative stress, inflammation, excitotoxicity of glutamate and imbalance of neurotrophic factors. In this paper, the research progress of DR nerve damage and its related factors are reviewed in order to provide new ideas for the prevention and treatment of DR.

Glia Connect Inflammation and Neurodegeneration in Multiple Sclerosis / 神经科学通报·英文版

Multiple sclerosis (MS) is regarded as a chronic inflammatory disease that leads to demyelination and eventually to neurodegeneration. Activation of innate immune cells and other inflammatory cells in the brain and spinal cord of people with MS has been well described. However, with the innovation of technology in glial cell research, we have a deep understanding of the mechanisms of glial cells connecting inflammation and neurodegeneration in MS. In this review, we focus on the role of glial cells, including microglia, astrocytes, and oligodendrocytes, in the pathogenesis of MS. We mainly focus on the connection between glial cells and immune cells in the process of axonal damage and demyelinating neuron loss.

Immunological Markers for Central Nervous System Glia / 神经科学通报·英文版

Glial cells in the central nervous system (CNS) are composed of oligodendrocytes, astrocytes and microglia. They contribute more than half of the total cells of the CNS, and are essential for neural development and functioning. Studies on the fate specification, differentiation, and functional diversification of glial cells mainly rely on the proper use of cell- or stage-specific molecular markers. However, as cellular markers often exhibit different specificity and sensitivity, careful consideration must be given prior to their application to avoid possible confusion. Here, we provide an updated overview of a list of well-established immunological markers for the labeling of central glia, and discuss the cell-type specificity and stage dependency of their expression.

Electroacupuncture Alleviates Functional Constipation in Mice by Activating Enteric Glial Cell Autophagy via PI3K/AKT/mTOR Signaling / 中国结合医学杂志

OBJECTIVE@#To investigate autophagy-related mechanisms of electroacupuncture (EA) action in improving gastrointestinal motility in mice with functional constipation (FC).@*METHODS@#According to a random number table, the Kunming mice were divided into the normal control, FC and EA groups in Experiment I. The autophagy inhibitor 3-methyladenine (3-MA) was used to observe whether it antagonized the effects of EA in Experiment II. An FC model was established by diphenoxylate gavage. Then the mice were treated with EA stimulation at Tianshu (ST 25) and Shangjuxu (ST 37) acupoints. The first black stool defecation time, the number, weight, and water content of 8-h feces, and intestinal transit rate were used to assess intestinal transit. Colonic tissues underwent histopathological assessment, and the expressions of autophagy markers microtubule-associated protein 1 light chain 3 (LC3) and Beclin-1 were detected by immunohistochemical staining. The expressions of phosphoinositide 3-kinases (PI3K)-protein kinase B (AKT)-mammalian target of rapamycin (mTOR) signaling pathway members were investigated by Western blot and quantitative reverse transcription-polymerase chain reaction, respectively. The relationship between enteric glial cells (EGCs) and autophagy was observed by confocal immunofluorescence microscopy, localization analysis, and electron microscopy.@*RESULTS@#EA treatment shortened the first black stool defecation time, increased the number, weight, and water content of 8-h feces, and improved the intestinal transit rate in FC mice (P<0.01). In terms of a putative autophagy mechanism, EA treatment promoted the expressions of LC3 and Beclin-1 proteins in the colonic tissue of FC mice (P<0.05), with glial fibrillary acidic protein (GFAP) and LC3 significantly colocalized. Furthermore, EA promoted colonic autophagy in FC mice by inhibiting PI3K/AKT/mTOR signaling (P<0.05 or P<0.01). The positive effect of EA on intestinal motility in FC mice was blocked by 3-MA.@*CONCLUSION@#EA treatment can inhibit PI3K/AKT/mTOR signaling in the colonic tissues of FC mice, thereby promoting EGCs autophagy to improve intestinal motility.

Correlation between retinal glial cells and proliferative vitreoretinopathy / 中华实验眼科杂志

Proliferative vitreoretinopathy (PVR) is a common complication of retinal detachment surgery, also an important blinding cause.Besides retinal pigment epithelial cells, retinal glial cells also are mainly involved in the formation and development of PVR.Retinal glial cells include Müller cells, astrocytes and microglia.The glial cells not only can be activated to change the morphology of cells, but also secrete growth factors and cytokines, synthesize extracellular matrix, and participate in the formation of PVR.The factors and synthetic extracellular matrix secreted by retinal glial cells promote the proliferation of retinal glial cells, which accelerates the development of PVR.This article described the interaction between the activation of retinal glial cells, astrocytes, growth factors secreted, cytokines secreted, extracellular matrix synthesized by glial cells and PVR formation, and the effect of non-coding RNAs on glial cells and PVR.

An optimized culture method of primary hippocampal neurons in SD rats / 中国药理学通报

Aim To establish a stable and efficient method for the primary culture of hippocampal neurons from serum-free fetal rats.so as to obtain hippoeampal neurons with higher purity and better vitality.Methods The hippocampal tissues of SI) rat fe¬tus rats on the ( 18 ± 1) day of pregnancy were cultured by Neu- robasal or DMEM/F12 medium and divided into serum-free cul-ture group, fetal bovine serum culture group and 5-fluoro-2'de- oxyuridine culture group.After cells of three groups were cul¬tured for 12 hours, all medium was changed to Neurobasal medi¬um.When the neurons in FUI)R culture group were cultured to 3 days.FLDR was added to inhibit the growth of glial cells.'Hie growing status of hippo cam pal neurons at different culture time points was observed,neuronal activity was detected,cell apopto¬sis was assessed, and the purity of hippocampal neurons was i- dentified.Results Compared with 10% serum culture group, the neurons cultured in serum-free culture group showed higher viability,lower apoptosis rate and higher purity; FUI)R reduced cell viability and increased cell apoptosis.Conclusions 'Hie neurons cultured by serum-free culture have good activity and high purity, instead of adding glial cell inhibitors, which is a fea¬sible and optimized primary culture method for hippocampal neu¬rons.

Research progress on the role of Müller glial cells in retinal nerve injury / 国际眼科杂志(Guoji Yanke Zazhi)

Müller glial cells(MGCs)are the major type of glial cells in the retina which radiating across the entire retina. MGCs make a close contact with retinal neurons, interact, and contribute to retinal homeostasis. After retinal nerve injury, MGCs respond to retinal injury in a variety of regulatory ways to protect the inner retinal environment changes and damage, producing retinal neuroprotective effects, such as regulating neurotransmitters release, releasing neuroprotective factors and antioxidant factors and reprogramming for endogenous repair. However, persistent pathological stimulation in retina can also exacerbate MGCs' proliferation which participate in neuronal dysfunction or loss. Therefore, a proper understanding of the response of MGCs to pathological stimuli and their protective and damaging effects will have a great impact on revealing mechanisms of retinal nerve damage disease and guiding the treatment of the disease. This article reviews the role of MGCs in retinal nerve injury and repair and provides new strategies for retinal neuroprotection.

Cultivation and characterization of primary human parathyroid cells from patients with severe secondary hyperparathyroidism / 南方医科大学学报

OBJECTIVE@#To establish an cell model of hyperparathyroidism by isolation, in vitro culture, and identification of parathyroid cells from patients with secondary hyperparathyroidism (SHPT).@*METHODS@#The parathyroid gland tissues obtained from 10 patients with SHPT were dissociated by collagenase digestion for primary culture of the parathyroid cells. Morphological changes and growth characteristics of the cells were assessed by microscopic imaging and cell counting. The mRNA and protein expression levels of parathyroid hormone (PTH), calcium-sensing receptor (CaSR), and glial cells missing 2 (GCM2) in the primary and passaged cells were determined by immunofluorescence, qRT-PCR, and Western blotting.@*RESULTS@#Primary cultures of parathyroid cells were successfully obtained. The cells exhibited a high expression of PTH shown by immunofluorescence assay and had a population doubling time of approximately 71.61 h. PTH secretion in the second-passage (P2) cells was significantly lower than that in the primary (P0) and first-passage (P1) cells (P < 0.001). Despite a significant downregulation of CaSR mRNA (P=0.017) and protein (P=0.006) in P1 cells as compared with P0 cells, no significant differences were found in mRNA and protein expressions of PTH or GCM2 between the two cell generations.@*CONCLUSION@#Primary cultures of parathyroid cells isolated from SHPT patients by collagenase digestion show similar biological properties to the cells in vivo.

Regulation of Naoxin'an Capsule on Glial Cell Activation and Inflammatory Response in Rats with Chronic Cerebral Hypoperfusion-induced Vascular Cognitive Impairment / 中国实验方剂学杂志

Objective:To investigate the protective effect of Naoxin'an capsule （NC） against glial cell activation and inflammatory damage in brain of rats with chronic cerebral hypoperfusion-induced vascular cognitive impairment （VCI）. Method:One hundred and fifty rats were randomly divided into a sham operation group （<italic>n</italic>=20） and a modeling group （<italic>n</italic>=130）. Following the modeling with the two vessels occlusion （2-VO） technique， 87 successfully modeled rats were randomly divided into the model group， positive drug group （aricept， 0.5 mg·kg^-1）， and low-， medium-， and high-dose （0.18， 0.36， 0.72 g·kg^-1） NC groups， with 17-18 rats in each group. After intragastric administration of NC for eight weeks， the Morris water maze test and passive avoidance test were conducted to detect the effects of NC on learning and memory ability of VCI rats. Changes in neuronal structure of rat hippocampal CA1 area were observed by hematoxylin-eosin （HE） staining， and the neuronal apoptosis in hippocampus by terminal deoxynucleotidyl transferase （TdT）-mediated dUTP nick end labeling （TUNEL） staining. Western blot assay was used to detect the expression levels of glial fibrillary acidic protein （GFAP）， ionized calcium-binding adapter molecule 1 （Iba-1）， phosphorylated p38 mitogen-activated protein kinase （p38 MAPK）， and phosphorylated nuclear factor <italic>κ</italic>B （NF-<italic>κ</italic>B）， followed by the measurement of interleukin-1<italic>β</italic> （IL-1<italic>β</italic>） and tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>） in the brain by enzyme-linked immunosorbent assay （ELISA）. Result:Compared with the sham operation group， the model group displayed obviously decreased spatial learning and memory ability and memory retention ability （<italic>P</italic><0.05， <italic>P</italic><0.01）， neuronal damage in hippocampal CA1 area， enhanced neuronal apoptosis （<italic>P</italic><0.01）， up-regulated GFAP and Iba-1 （<italic>P</italic><0.01）， elevated phosphorylation of p38 MAPK and NF-<italic>κ</italic>B （<italic>P</italic><0.01）， and increased IL-1<italic>β</italic> and TNF-<italic>α</italic> （<italic>P</italic><0.01）. Compared with the model group， NC at each dose significantly improved the spatial learning and memory ability and memory retention ability of VCI rats （<italic>P</italic><0.05， <italic>P</italic><0.01）， ameliorated the neuronal damage in hippocampus CA1 area， reduced the apoptosis rate of nerve cells （<italic>P</italic><0.05， <italic>P</italic><0.01）， down-regulated the expression of GFAP and Iba-1 （<italic>P</italic><0.01）， decreased the phosphorylation levels of p38 MAPK and NF-<italic>κ</italic>B （<italic>P</italic><0.05， <italic>P</italic><0.01）， and lowered TNF-<italic>α</italic> and IL-1<italic>β</italic> levels （<italic>P</italic><0.01）. Conclusion:NC alleviates the inflammatory damage of the central nervous system caused by activated p38 MAPK and NF-<italic>κ</italic>B and improves chronic cerebral hypoperfusion-induced VCI in rats by inhibiting the activation of microglia and astrocytes.

Research of Glial Cell Line-derived Neurotrophic Factor in Gastrointestinal Diseases / 胃肠病学

Glial cell line-derived neurotrophic factor (GDNF) is an important functional protein derived from enteric glial cells. It plays an important role in the enteric nervous system, such as nutritional neurons, promoting synaptic remodeling and anti-inflammation. The role of GDNF in the progression of gastrointestinal diseases has received more attention gradually. This article reviewed GDNF and its ligands, related signaling pathway, correlation with intestinal homeostasis and clinical application.

Research progress of Müller cells and retinal nerve regeneration / 国际眼科杂志(Guoji Yanke Zazhi)

@#Retinal degenerative diseases, a type of blinding eye diseases in which retinal neuron apoptosis is the main pathological process. Neuronal cells cannot be regenerated after damage, Müller cells are important glial cells of the retina and involved in retinal development, damage, and regeneration process. In recent years, studies have proved that Müller cells are an endogenous alternative source for stimulating damaged retinal neurons and an excellent target for retinal nerve regeneration. This article reviews the related factors of Müller cells and retinal nerve regeneration, and provides a new direction for nerve regeneration research.

Advance in Development of Neurons and Glial Cells in Brain Tissue of Children with Autism Spectrum Disorder (review) / 中国康复理论与实践

Autism spectrum disorder is a group of neurodevelopmental disorders that occur in early childhood. The specific pathogenesis is not clear. Abnormal development of neurons and glial cells is a significant feature of patients with autism spectrum disorder. This article reviewed the changes in neurons, astrocyte, microglial cell and oligodendroglia cell in autism spectrum disorder.

Effect of electroacupuncture on recognition memory and levels of Aβ, inflammatory factor proteins and aquaporin 4 in hippocampus of APP/PS1 double transgenic mice / 针刺研究

OBJECTIVE: To investigate the effect of electroacupuncture (EA) at "Baihui "(GV20) and "Shenshu "(BL23) on activation of glial cells, expression of inflammatory factor proteins and aquaporin 4 (AQP4)in the hippocampus of amyloid precursor protein/presenilin-1 (APP/PS1) transgenic mice, so as to explore its mechanisms underlying improvement of Alzheimer's disease(AD). METHODS: Twenty C57/BL6 background male APP695/PS1-dE9(APP/PS1) double transgenic mice (model group) and 20 wild type (WT) C57/BL6 mice (blank group) were respectively randomized into control and EA groups. EA (2 Hz/15 Hz, 1－2 mA) was applied to GV20 and bilateral BL23 for 30 min, once daily, 6 days a week for 4 weeks. The recognition memory ability was detected by novel object recognition tests in a behavior test box. The percentage of time spent in close interaction with novel object (C) relative to the total time was used to generate preference index. The contents of hippocampal β amyloid protein (Aβ)1-40 and Aβ1-42 were assayed using ELISA, and the expression levels of glial fibrillary acidic protein (GFAP), ionic calcium binding receptor molecule-1 (Iba-1), interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) proteins in the hippocampus measured by Western blot. The activities of hippocampal astrocytes (GFAP-labelled cells), microglia (Iba-1-labelled cells) and the polarity expression of AQP4 (for removing Aβ) were measured by immunohistochemistry. RESULTS: The preference index was significantly decreased in the model group relatively to the blank control group (P0.05)．. CONCLUSION: EA at GV20 and BL23 can reduce inflammatory reaction and Aβ level, suppress activation of astrocytes and microglia, and up-regulate expression of AQP4 in the hippocampus tissue in APP/PS1 transgenic mice, which may contribute to its effect in improving recognition memory ability, suggesting a role of EA intervention in delaying the development of AD via promoting the drainage of Aβ by the glymphatic system in the brain.

Study on the Improvement Effects of Dajianzhong Decoction on Irritable Bowel Syndrome Visceral Pain Model Rats Based on EGC / 中国药房

OBJECTIVE：To study th e improvement effect and mechanism of Dajianzhong decoction on irritable bowel syndrome（IBS）visceral pain model rats. METHODS ：Totally 48 male non-weaning rats were randomly divided into normal control group ，model group ，pinaverium bromide group （positive control ，45 mg/kg）and Dajianzhong decoction high-dose ， medium-dose and low-dose groups （2.16，1.08，0.54 g/kg，by crude drug ），with 8 rats in each group. Except for normal control group，IBS visceral pain model was established by mother and child separation ，acetic acid enema ，ovalbumin intraperitoneal Remote limb precondition - ing protects against ischemia induced neuronal death through ameliorating neuronal oxidative DNA damage injection in other groups for 57 d. On the 58th day ，the rats in administration groups were given the corresponding drugs intragastrically，and model group and normal control group were given constant volume of purified water ，once a day ，for consecutive 14 d. The general condition of rats was observed ；abdominal wall withdrawal reaction （AWR）was adopted to evaluate the visceral sensitivity of rats in each group under 20，40，60，and 80 mmHg （1 mmHg＝0.133 kPa）pressure；HE staining method was used to observe the colon pathological features of rats in each group. Western blotting assay was used to detect the protein expression of intestinal glial cells markers fibrillary acidic protein （GFAP），nerve growth factor （NGF）and its receptor TrkA in colon tissue. RESULTS ：The mucosal layer of colon tissue in rats of model group was discontinuous ，gland edema was observed and lymphocytes ，neutrophils and eosinophils were scattered in the lamina propria. In Dajianzhong decoction low-does group，the mucosal layer of colon tissue was incomplete ，some glands were slightly edematous ，and a few lymphocytes ， neutrophils in the lamina propria. The colonic mucosa epithelial structure was intact ，glands arranged regularly ，and no degenerative necrosis and inflammatory cells were observed in Dajianzhong decoction medium- and high-dose groups and pinaverium bromide group. Compared with normal control group ，AWR scores under 20，40，and 60 mmHg pressure ，relative protein expression of GFAR ，NGF and TrkA were all increased significantly in model group and Dajianzhong decoction low-does groups（P＜0.05 or P＜0.01）. Compared with model group ，AWR scores under 20，and 40 mmHg pressure in Dajianzhong decoction medium- and high-dose groups and pinaverium bromide group ，AWR scores under 60 mmHg pressure in Dajianzhong decoction high-dose group and pinaverium bromide group ，relative protein expression of GFAP ，NGF，TrkA in colon tissue in Dajianzhong decoction medium- and high-dose groups and pinaverium bromide group were all decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS ：Dajianzhong decoction could improve the visceral pain of IBS model rats by inhibiting the activation of intestinal glial cells and reducing the expression of NGF and TrkA.

Regulation and mechanism of hypothalamic inflammation on obesity / 中华内分泌代谢杂志

Many studies of the etiology and intervention for obesity have gradually focused on the brain, trying to curb the occurrence of obesity from the source. Hypothalamic inflammation has been a concern and an unresolved scientific issue in the development of obesity. Studies have shown that hypothalamic inflammation not only impairs energy balance, but also increases obesity-related insulin and leptin resistance, further promotes peripheral tissues storing up fat cells, eventually leads to the development of obesity. In addition, hypothalamic inflammation occurs before weight-gain and peripheral tissue inflammation with high-fat diets. Therefore, more and more scholars believe that hypothalamic inflammation is an important cause of dietary-induced metabolic abnormalities. The occurrence of hypothalamic inflammation is mainly accompanied by a series of complex and rapidly-activated glial, including microglia, astrocytes, and tanycyte. These cells are responsible for maintaining hypothalamic metabolic homeostasis and making up the important components of the regulatory network. Moreover, multiple teams also found that a variety of weight-loss methods(e.g. bariatric surgery, targeted drugs, fecal microbiota transplantation, and so on) can improve hypothalamic inflammation levels. Therefore, it is important to understand the mechanism of hypothalamic inflammation through different neurons, which is expected to find a more effective and safer solution to intervene and treat obesity in the future.

Postmortem analysis of vagus nerve pathology and intestinal neuroinflammation in Parkinson's disease / 中华神经科杂志

Objective To investigate the roles of neuroinflammation and pathology of peripheral nervous system in the pathogenesis of Parkinson's disease (PD).Methods Immunohistochemical staining was performed to examine the nodose ganglia (containing vagus nerve) and intestine of an autopsy case of PD.The neuroinflammation and morphological changes of vagus nerve and enteric nervous system were observed.Results The expressions and distributions of glial fibrillary acidic protein and ionized calcium binding adapter molecule 1,two typical glia cell biomarkers,were different in vagus nerve and intestinal mucosa.Tumor necrosis factor α and inducible nitric oxide synthase were expressed in intestinal mucosa and myenteric plexus of the PD patient.There was a strong inflammatory reaction in the myenteric plexus,which distributed diffusely.Conclusion Satellite glial cells and intestinal inflammatory response may play a role in the pathogenesis of PD.

Expression and significance of HA/CD44 in glial cells and optic cross nerves / 国际眼科杂志(Guoji Yanke Zazhi)

@#Hyaluronic acid(HA)is one of the main components of the extracellular matrix(ECM), and it is participated in many cells physiology and pathological processes, such as tissue reconstruction, expansion of cell gap, inflammation and tumorigenesis and so on. CD44 is a cell surface receptor for HA and widely distributed cell surface glycoprotein, which paticipate in specific adhesion of cell to cell and cell to matrix. CD44 is the most important hyaluronic acid receptor on the cell surface. Besides, CD44 is the main site of binding to HA. In this paper, we will elaborate from three aspects: the binding of HA and CD44 and its molecular basis, the expression and significance of HA/CD44 in glial cells(including Müller cells)and the expression and significance of HA/CD44 in the optic nerve, which makes readers have an understanding of the role of HA and CD44.

Glial response following diffuse axonal injury / 解剖学报

Objective To explore the glial response and the relationship with secondary axonal degeneration in rats after diffuse axonal injury (DAI). Methods Adult male Sprague-Dawley rats were randomly assigned to control or DAI groups sacrificed at 1, 2, 3, 5 and 7 days with 10 rats in each group. DAI model was made referring to modified Marmarou method and glial fibrillary acidic protein (G F A P), ionized calcium binding adaptor molecule-1 (Ibal), recombinant oligodendrocyte lineage transcription factor 2 (Olig2), CC-1, NG2 immunohistochemistry, TUNEL staining and transmission electron microscopy were performed in brain stem. Results The number of Ibal labeled positive cells was significantly increased at day 3 and day 7 after injury. Moreover, different hypertrophic morphology was identified after injury. There was no effect of DAI on GFAP expression in brain stem. Numbers of mature oligodendrocyte marker CC-1 immunoreactivity cells within brain stem were significantly decreased at each of the time points after injury. The number of TUNEL positive cells in brain stem was significantly increased with injured time. Olig2 expression was significantly increased throughout the first week and reached peak at day 3 after injury in brain stem. The number of NG2 labeled positive cells was significantly increased at day 3 and day 7 after DAI. Ultrastructural evidence showed myelin release then further developed as widespread delamination and collapse, and leading to degeneration of axonal partner. Conclusion Mature oligodendrocytes are vulnerable in DAI and myelin loss may contribute to axonal degeneration. OPCs proliferate with activation of microglia. This insight of glial response will further explain the pathophysiological mechanism of secondary axonal damage in DAI.

A new perspective reexploration of neonatal mice glial culture in vitro and inflammatory model / 中国药理学通报

Aim To revisit the long-established murine glial cul-ture-base inflammation model, in order to explore the possible improvements of current methods. Methods The proportion and purity of glial cells were characterized by qPCR, flow cytometr) and immunofluorescence. After stimulation with LPS, their inflammation response was evaluated at both mRNA and protein levels. Results Mixed glial cells were stimulated by LPS (IOC jig L-1) , and the expression of inflammatory factors increased more significantly than that of microglial. For example, at PDL-coated condition, TNF-a increased more significantly in mixec glial cells ( 903. 8 ± 322. 2 ) ng L-1 than that in microglia (565.4 ± 159. 8)ng • L-1. When cultured as a glial mixture, cells grew better on a matrigel-coated surface. However, when cultured in uncoated condition, purified microglia were more sensitive to LPS [TNF-ot; (6861.4 ± 1606.6) ng L-1[. Conclusions Matrigel, as a newly emerged coating material, is proved to be better than the conventional PDL-coating for cultu-ring mixed glial cells, which improves cell viability and sensitivity. Multiplex inflammatory factor assays should be used for quantifying inflammatory response, rather than relying on qPCR alone.