Ginkgolide B Promotes Neural Function Recovery of Ischemic Stroke Mice by Regulating Characteristics of Brain T Cells and Their Interactions with Glial Cells / 实验动物与比较医学

ObjectiveTo investigate the regulatory effects of Ginkgolide B on the biological characteristics of brain T cells and their interactions with glial cells during the recovery phase of ischemic stroke in mice. Methods36 adult C57BL/6 mice were randomly assigned to three groups: sham-operated group (Sham group), control group (PBS group), and Ginkgolide B treatment group (GB group). The Sham group underwent only sham surgeries, whereas the PBS and GB groups were subjected to a middle cerebral artery occlusion (MCAO) model using the filament method, followed by intranasal administration of an equivalent volume of either PBS or Ginkgolide B solution for 14 days post-injury. Neurological function changes were evaluated in all three groups using the rotarod test and a neurological scoring system. On day 15, single-cell sequencing was performed on fresh tissues from the brain injury areas, surrounding cortex, corpus callosum, and striatum of mice in the PBS and GB group to assess the biological characteristics of T cells and their subpopulations, and further explore the interactions and mechanisms among T cells, microglia, and oligodendrocytes. ResultsCompared with the Sham group, both PBS and GB group exhibited significant improvements in neurological scores and reduced pre-fall motor durations (P < 0.001). Compared with the PBS group, the GB group showed a downward trend in neurological scores and an upward trend in pre-fall motor durations on days 5, 10, and 15 post-ischemic brain injury, with a significant increase in pre-fall motor duration on day 15 (P < 0.05). Compared with the PBS group, the GB group exhibited a significant increase in T cell proliferative activity in the brain 15 days post brain injury (P < 0.05). The number of proliferative T cells and the levels of lipid metabolism were significantly elevated (P < 0.05), and there was a significant increase in extracellular matrix remodeling in all T cells (P < 0.05). Additionally, the interactions between T cells and both microglia and oligodendrocytes, as well as among the microglia themselves and between microglia and oligodendrocytes, were significantly enhanced in the GB group. This was primarily evident in the strengthened interactions between CD74 and macrophage migration inhibitory factor (MIF), as well as colony stimulating factor 1 receptor (CSF1R) and colony stimulating factor 1 (CSF1) (P < 0.05). However, the inflammatory levels of T cells showed no significant differences compared with the PBS group. ConclusionA mouse model of ischemic stroke can be successfully established by MCAO operation. Ginkgolide B may promote neurological recovery post-brain injury in mice by modulating the biological characteristics of T cells within the brain and their interactions with glial cells.

Effects of early postnatal immune activation on autism-like behaviors and glial cells in mice / 中国儿童保健杂志

【Objective】 To observe the effects of early postnatal immune activation (EPIA) on social behaviors of male and female mice, and to explore the possible role of the functional state of astrocytes and microglia in this process. 【Methods】 Using lipopolysaccharide (LPS) induced EPIA mice as study subjects, mice were divided into the male-control, male-model, female-control, and female-model groups, each containing 10 mice (n=10). Behavioral tests were performed at 25 - 32 days of age, and the social behavior ability of mice was evaluated by open field test, three-chamber sociability test, and marble burying test. The expression levels of GFAP, IBA-1, TLR4, and NFκB p65 in the cortex and hippocampus were detected by Western blot (n=3). 【Results】 In behavioral tests, social index significantly decreased in LPS treatment group (F=14.907, P<0.05). The interaction effect between treatment and sex was significant in the residence time (F=5.260, P<0.05) and the number of buried marbles (F=7.788, P<0.05). LPS treatment decreased the retention time of the central region in male mice (F=4.261, P<0.05), and increased the number of buried marbles in males (F=20.645, P<0.05). Western blot results showed that LPS treatment increased the expression of GFAP protein in the hippocampus (F=50.443, P<0.05) and cortex (F=30.116, P<0.05), as well as the expression of IBA-1 protein (F=21.844. P<0.05) and TLR4 protein (F=6.215, P<0.05) in the cortex. The results of NFκB p65 showed a significant interaction between treatment and sex in the cortex (F=6.558, P<0.05), and LPS increased the expression of NFκB p65 protein in the cortex in female mice (F=16.317, P<0.05). 【Conclusions】 EPIA is sufficient to induce sex-specific autism spectrum disorder (ASD)-like behavior and enhance astroglial and microglial reactivity in mice. ASD-like behavior induced by EPIA may be related to the TLR4/NFκB signaling pathway in the cortex.

The Role of Prefrontal Cortex in Social Behavior / 生物化学与生物物理进展

Social behavior is extremely important for the physical and mental health of individuals, their growth and development, and for social development. Social behavioral disorders have become a typical clinical representation of a variety of psychiatric disorders and have serious adverse effects on the development of individuals. The prefrontal cortex, as one of the key areas responsible for social behavior, involves in many advanced brain functions such as social behavior, emotion, and decision-making. The neural activity of prefrontal cortex has a major impact on the performance of social behavior. Numerous studies demonstrate that neurons and glial cells can regulate certain social behaviors by themselves or the interaction which we called neural microcircuits; and the collaboration with other brain regions also regulates different types of social behaviors. The prefrontal cortex (PFC)-thalamus projections mainly influence social dominance and social preference; the PFC-amygdala projections play a key role in fear behavior, emotional behavior, social exploration, and social identification; and the PFC-nucleus accumbens projections mainly involve social preference, social memory, social cognition, and spatial-social associative learning. Based on the above neural mechanism, many studies have focused on applying the non-invasive neurostimulation to social deficit-related symptoms, including transcranial magnetic stimulation (TMS), transcranial electrical stimulation (TES) and focused ultrasound stimulation (FUS). Our previous study also investigated that repetitive transcranial magnetic stimulation can improve the social behavior of mice and low-intensity focused ultrasound ameliorated the social avoidance behavior of mice by enhancing neuronal activity in the prefrontal cortex. In this review, we summarize the relationship between neurons, glial cells, brain projection and social behavior in the prefrontal cortex, and systematically show the role of the prefrontal cortex in the regulation of social behavior. We hope our summarization will provide a reference for the neural mechanism and effective treatment of social disorders.

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.

Comparison of the infectivity and stability of Asian versus African lineage Zika virus in glial cells / 中国人兽共患病学报

Infection of pregnant women by Asian lineage Zika virus(ZIKV)is more likely to cause microcephaly in infants than African lineage ZIKV.To further clarify the differences in infectivity and stability between Asian and African lineage ZIKV,we selected Asian lineage SZ01 and African lineage MR766 strains for study in glial cells.First,we examined the differ-ence in infectivity between SZ01 and MR766 on U251 and U87 astrocytes,and T98G glioblastoma cells,by using CCK8 assays.Subsequently,we examined the tolerance of SZ01 and MR766 to 37 ℃ and 40 ℃ in the free and infected cell states,by using qRT-PCR or viral plaque assays.Finally,we examined the effect of repeated freezing and thawing on the stability of SZ01 and MR766 with viral plaque assays.ZIKV had higher infectivity in U251 and U87 than T98G,and SZ01 was more infectious to as-trocytes than MR766.SZ01 tolerated 40 ℃ better than MR766 in the free state.SZ01 proliferated faster than MR766 in glial cells at 40 ℃.The tolerance of SZ01 to repeated freezing and thawing was higher than that of MR766.The above findings sug-gest that the infectivity and stability of Asian lineage ZIKV are significantly higher than those of African lineage ZIKV,possi-bly because of its persistent infection and pathogenicity.

Intrathecal administration of Maresin 1 attenuates bone cancer pain by inhibiting the glial cells acti-vation and neuroinflammation in the spinal cord / 临床麻醉学杂志

Objective To investigate the effects of intrathecal administration of Maresin 1 on pain behavior and glial cells activation in spinal dorsal horn of mice with bone cancer pain(BCP).Methods Experiment Ⅰ,twenty-eight clean male C57BL/6 mice,aged 4-6 weeks,weighing 18-22 g,were ran-domly divided into two groups:sham 1 group(group S1)and BCP 1 group(group B1),14 mice in each group.BCP model was established in group B1 by injecting PBS 20 μl containing 2 × 105 lewis lung carcino-ma cells into the intramedullary space.Group S,received PBS 20 μl without tumor cells.Six mice in each group were randomly selected to determine the mechanical withdrawal threshold(MWT)and thermal with-drawal latency(TWL)1 day before implantation,3,7,14,21 days after implantation.Four mice were randomly sacrificed on days 7,14 and 21 in group S,and group B1.The expressions of spinal glial fibrillary acidic protein(GFAP)and ionized calcium-binding adapter molecule 1(Iba1)were detected by Western blot.Experiment Ⅱ,thirty-six clean male C57BL/6 mice were randomly divided into three groups:sham 2 group(group S2),BCP 2 group(group B2),and Maresin 1 group(group M),12 mice in each group.BCP models were established in groups B2 and M while group S2 received PBS 20 μl without tumor cells.Maresin 1 50 ng/5 μl were intrathecally injected from days 14 to 16 after tumor cells implantation.Six mice in each group were randomly selected to determine the MWT and TWL on days 14 to 18 after implantation.Mice were sacrificed on day 18 after pain behavior tests.The expressions of spinal GFAP and Iba1 were de-tected by Western blot and immunofluorescence staining.The levels of spinal IL-1 β,IL-6,and TNF-α were detected by ELISA.Results Experiment Ⅰ:compared with group S1,the MWT and TWL were significantly lower on days 7,10,14,and 21 after implantation in group B,(P＜0.05),the expressions of spinal GFAP and Iba1 were upregulated on days 7,14,and 21 after implantation in group B1(P＜0.05).Experiment Ⅱ:compared with group S2,the levels of spinal GFAP,Iba1,IL-1β,IL-6,and TNF-α were increased on day 18 after implantation in group B2(P＜0.05).Compared with group B2,the MWT and TWL were significantly higher on days 15-18 after implantation while levels of spinal GFAP,Iba1,IL-1β,IL-6,and TNF-α on day 18 were decreased in group M(P＜0.05).Conclusion Intrathecal administra-tion of Maresin 1 attenuates BCP maybe by inhibiting the glial cells activation and neuroinflammation in the spinal cord.

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.

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.

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.

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.

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.

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.

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.

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.

The role of α7-nAChR /PI3K/AKT/GSK-3 pathway in chronic sleep deprivation and its mechanism / 中风与神经疾病杂志

@#Objective Investigated the influences of chronic sleep deprivation on the expression α7-nAChR and the activity of its downstream PI3K/AKT/GSK-3β pathway in neuroglia in mice.Methods Adult C57BL/6J mice aged 7-8 weeks were randomly divided into three groups，control（CC）group，chronic sleep deprivation（SD） group，chronic sleep deprivation + intraperitoneal injection α7-nAChR agonists PHA-543613（SD + PHA - 543613） group.The protein expression of α7 -nAChR,p-AKT,p-GSK-3β,Nrf-2,HO-1 in mice hippocampus were examined by Western Blot，the gene expression of α7 -nAChR and the gene expression of TNF-α,IL-1β,IFN-γ,MCP-1,Arg-1,CD206,TGF-β,YM-1 were examined by Rt-PCR；the expression of α7-nAChR on astrocytes and microglias in mice hippocampus were assesed by double-labeled immunofluorescence.Spatial learning and memory was assessed by morris water maze （MWM） test.Results After chronic sleep deprivation for 7 days，the expression of α7-nAChR protein，mRNA in SD group decreased significantly when compared with CC group（P=0.001,P=0.038）;the expression of protein p-AKT decreased in SD group compared with CC group（P=0.019）,while the expression of protein p-GSK-3β increased in SD group compared with CC group（P=0.011）.Afterα7-nAChR agonists PHA-543613 treatment,the expression of α-7 nAChR on astrocytes increased significantly when compared with SD group（P=0.027，the expression of p-AKT and p-GSK-3βincreased when compared with SD group（P=0.047,P=0.017.At the same time，the expression of Nrf-2，HO-1 and anti-inflammatory factors（CD206，TGF-β）increased significantly compared with SD group（P=0.020,P=0.016，P<0.01,P<0.01），while the expression of inflammatory factors（TNF-α，MCP-1） decreased compared with SD group（P<0.01）.After α7-nAChR agonists PHA-543613 treatment，the escape latency at the end of experiment was decreased，the frequency of crossing platforms and the time in the target quadrant was prolonged compared to the SD group（P=0.000,P=0.000,P=0.001）.Conclusion Stimulation of α7-nAChR reduces inflammation and oxidative stress via activation of PI3K/AKT /GSK-3β after chronic sleep deprivation.

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.

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.

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.

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.