• Title, Summary, Keyword: Reactive astrocytes

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Effect of Glial-neuronal Cell Co-culture on GFAP Expression of Astrocytes (신경세포가 별아교세포의 아교섬유성 산단백질 표현에 미치는 영향)

  • Bae Hyung-Mi;Park Jung-Sun;Yeon Dong-Soo
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.3
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    • pp.285-296
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    • 1997
  • Injury to brain transforms resting astrocytes to their reactive form, the hallmark of which is an increase in glial fibrillary acidic protein (GFAP), the major intermediate filament protein of their cell type. The overall glial response after brain injury is referred to as reactive gliosis. Glial-neuronal interaction is important for neuronal migration, neurite outgrowth and axonal guidance during ontogenic development. Although much attention has been given to glial regulation of neuronal development and regeneration, evidences also suggest a neuronal influence on glial cell differentiation, maturation and function. The aim of the present study was to analyze the effects of glial-hippocampal neuronal co-culture on GFAP expression in the co-cultured astrocytes. The following antibodies were used for double immunostaining chemistry; mouse monoclonal antibodies for confirm neuronal cells, rabbit anti GFAP antibodies for confirm astrocytes. Primary cultured astrocytes showed the typical flat polygonal morphology in culture and expressed strong GFAP and vimentin. Co-cultured hippocampal neurons on astrocytes had phase bright cell body and well branched neurites. About half of co-cultured astrocytes expressed negative or weak GFAP and vimentin. After 2 hour glutamate (0.5 mM) exposure of glial-neuronal co-culture, neuronal cells lost their neurites and most of astrocytes expressed strong CFAE and vimentin. In Western blot analysis, total GFAP and vimentin contents in co-cultured astrocytes were lower than those of primary cultured astrocytes. After glutamate exposure of glial-neuronal co-culture, GFAP and vimentin contents in astrocytes were increased to the level of primary cultured astrocytes. These results suggest that neuronal cell decrease GFAP expression in co-cultured astrocytes and hippocampal neuronal-glial co-culture can be used as a reactive gliosis model in vitro for studying GFAP expression of astrocytes.

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Increased CNTF Expression in the Reactive Astrocyte Following Spinal Cord Injury in Rats (흰쥐에서 척수 손상후 반응성 별아교세포에서의 CNTF 발현 증가)

  • Kim, Chang-Jae;Moon, Se-Ho;Lee, Byung-Ho;Chung, Mee-Young;Chea, Jun-Seuk;Lee, Mun-Yong;Chun, Myung-Hoon
    • The Korean Journal of Pain
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    • v.11 no.2
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    • pp.182-193
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    • 1998
  • Background: Ciliary neurotrophic factor (CNTF), identified as a survival factor for developing peripheral neurons is upregulated by reactive astrocytes in the traumatized tissue and in areas of terminal degeneration after a brain lesion. But in the spinal cord, CNTF is expressed in the non-astrocytic phenotypic, maybe oligodendrocytes. The present study was undertaken to determine the upregulation of CNTF expression in reactive astrocytes following spinal cord lesion in the rat. Methods: Unilateral incision of the dorsal funiculus at the thoracic level was performed and rats were sacrificed on days 3, 7, 14 and 28 postlesion. Western blot analysis, immunocytochemical analysis and double immunofluorescence for CNTF and glial fibrillary acidic protein (GFAP) were performed after spinal cord lesion. Results: A major band with 24 kDa and additional band of higher molecular weight form were detectable, and the intensity of the 24 kDa immunoreactive band increased up to 14 days postlesion and decreased toward laminectomized control values. CNTF immunoreactivity was markedly upregulated in the injured dorsal funiculus and adjacent gray matter. The time course of CNTF expression is coincident with the appearance of reactive astrocytes in the injured spinal cord. Moreover, double immunofluorescence for CNTF and glial fibrillary acidic protein (GFAP) revealed that CNTF immunoreactivity was in GFAP immunoreactive astrocytes. Conclusions: These results show that CNTF upregulation occurred in reactive astrocytes following spinal cord lesion, and suggest a role for CNTF in the regulation of astrocytic responses after spinal cord injury.

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Cytotoxic Effect of Free Radical on Rat Primary Astrocytes (자유라디칼이 백서의 뇌별아교세포에 미치는 독성작용)

  • Jang, Hyuk;Kim, Myung-Sunny;Park, Hyun-Young;Kim, Yo-Sik;Cho, Kwang-Ho;Chung, Hun-Taeg;Park, Rae-Kil
    • Toxicological Research
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    • v.16 no.1
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    • pp.1-8
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    • 2000
  • Astrocytes generate free radicals including nitric oxide (NO) and reactive oxygen intermediates(ROI) which in turn play roles in the pathogenesis of degenerative diseases and sclerotic changes of the brain. This study was designed to evaluate the mechanism that free radicals contribute to the cytotoxicty of rat neonatal primary astrocytes. Treatment with NO donors alone including soldium nitroprusside(SNP), S-nitrosoglucathinoe (GSNO), and S-nitroso-n-acetylpenicillamine (SNAP) showed a little effect on the death of rat neonatal primary astrocytes, whereas SNP markedly induced the death of RAW 264.7 cells. ROI inculding H2O2 and O2 donor also slightly induced the death of rat primary astrocytes. However, 3-morpholinosydnonimine(SIN-1), a donor of peroxynitrite (ONOO), which is a reactive compound of NO with superoxide, significantly decreased the viability of rat primary astrocytes in a dose-dependent manner. Cells were retarded in outgrowth of viability of cellular processes with cell shrinkage and detachment from culture dishes. Hoechst staining demonstrated that SIN-1-induced cell death might be due to an apoptosis which was characterized by nuclear condensation and fragmentation. SIN-1-induced apoptosis was prevented by the pretreatment with superoxide dismutase (SOD) and catalase in rat primary astorocytes. Furthermore, prevention of the generation of reduced glutathione (GSH) by DL-buthionine-[S, R]-sulfoximine (BSO) aggravated the cytotoxic effects of SNP, benzene triol, and SIN-1 in rat primary astrocytes. Taken together, it is suggested that peroxynitrite may be a major effector of apoptosis and cellular antioxidant system is important for cell survival in rat prima교 astrocytes.

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Effect of acupuncture on short-term memory and apoptosis after transient cerebral ischemia in gerbils

  • Choi, In-Ho;Lim, Hyung-Ho
    • The Journal of Korean Medicine
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    • v.39 no.4
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    • pp.1-15
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    • 2018
  • Objectives: Cerebral ischemia results from a variety of causes that cerebral blood flow is reduced due to a transient or permanent occlusion of cerebral arteries. Reactive astrocytes and microglial activation plays an important role in the neuronal cell death during ischemic insult. Acupunctural treatment is effective for symptom improvement in cerebrovascular accident, including cerebral ischemia. Methods: In the present study, the effects of acupuncture at the ST40 acupoint on short-term memory and apoptosis in the hippocampal CA1 region following transient global cerebral ischemia were investigated using gerbils. Transient global ischemia was induced by occlusion of both common carotid arteries with aneurysm clips for 5 min. Acupuncture stimulation was conducted once daily for 7 consecutive days, starting one day after surgery. Results: In the present results, ischemia induction deteriorated short term memory, increased apoptosis, and induced reactive astrocyte and microglial activation. Acupuncture at ST40 acupoint ameliorated ischemia-induced short-term memory impairment by suppressing apoptosis in the hippocampus through down-regulation of reactive astrocytes and microglial activation. Conclusion: The present study suggests that acupuncture at the ST40 acupoint can be used for treatment of patients with cerebral stroke.

A Study on the Effects of Sunghyangjungkisan-ga-pogokyoung on In vitro Alzheimer's Disease Experimental Model (생체외(生體外) 알츠하이머병 실험(實驗) 모델에서 성향정기산가포공영(星香正氣散加蒲公英)의 효과(效果)에 관(關)한 연구(硏究))

  • Kang Hyung-Won;Lyu Yeoung-Su;Park Jin-Sung
    • Journal of Oriental Neuropsychiatry
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    • v.12 no.2
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    • pp.157-171
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    • 2001
  • Astrocytes are glial cells that play a major role in the inflammation observed in Alzheimer's disease (AD). Upon stimulation from various agents, these cells adopt a reactive phenotype, a morphological hallmark in AD pathology, during which they themselves may produce still more inflammatory cytokines. Substance P (SP) can stimulate secretion of tumor necrosis $factor-\;{\alpha}$ $(TNF-\;{\alpha})$ from astrocytes stimulated with lipopolysaccharide (LPS). Here I report that Sunghyangjungkisan- ga- pogokyoung(Sgp) can modulate cytokines secretion from primary cultures of rat astrocytes. Sgp $(10\;to\;1000\;{\mu}g/ml)$ significantly inhibited the $TNF-\;{\alpha}$ secretion by astrocytes stimulated with LPS and SP. Interleukin-1 (IL-1) has been shown to elevate $TNF-\;{\alpha}$ secretion from LPS-stimulated astrocytes while having no effect on astrocytes in the absence of LPS. Treatment of Sgp $(10\;to\;1000\;{\mu}g/ml)$ to astrocytes stimulated with both LPS and SP decreased IL-1 secretion significantly. The secretion of $TNF-\;{\alpha}$ by LPS and SP in astrocytes was progressively inhibited with increasing amount of IL-1 neutralizing antibody. Neurodegenerative processes in AD are thought to be driven in part by the deposition of ${\beta}\;-amyloid\;(A\;{\beta})$, a 39- to 43-amino acid peptide product resulting from an alternative cleavage of amyloid precursor protein. Sgp $(10\;to\;1000\;{\mu}g/ml)$ significantly inhibited the $TNF-\;{\alpha}$ secretion by astrocytes stimulated with $A-{\beta}-$and IL-1. These results suggest that Sgp may inhibit $TNF-\;{\alpha}$ secretion by inhibiting IL-1 secretion and that Sgp has an antiinflammatory activity in AD brain

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Modulation of LPS-Stimulated Astroglial Activation by Ginseng Total Saponins

  • Kim, Sok-Ho;Shim, Se-Hwan;Choi, Dea-Seung;Kim, Jong-Hoon;Kwon, Young-Bae;Kwon, Jung-Kee
    • Journal of Ginseng Research
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    • v.35 no.1
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    • pp.80-85
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    • 2011
  • Ginseng, a traditional medicine in Asian countries, is known to prevent various neuropathologic diseases such as Alzheimer's. Ginseng total saponins (GTS) in particular are one of the most effective ginseng extract compounds for neuroprotection. However, their protective effects on astrocytes are rarely reported. In pathological circumstances, astroglial activation plays a pivotal role in neuroinflammation. Subsequently, neuroinflammation induced by activated astrocytes causes brain damage. The purpose of the present study was to determine the suppressive effects of GTS on astroglial activation in lipopolysaccharide (LPS)-stimulated rat primary astrocytes. Astrocytes treated for 24 h with LPS demonstrated suppressed glialfibrillary acidic protein expression in a dose-dependent manner in the presence of GTS. GTS reduced production of proinflammatory cytokines such as tumor necrosis factor-${\alpha}$ and interleukin-1${\beta}$ and inhibited the level of inducible nitric oxide synthase, and cyclooxygenase-2 in LPS-stimulated astrocytes. Furthermore, GTS suppressed intracellular reactive oxygen species production. These modulations due to GTS may indicate neuroprotective antiinfl ammatory properties which may in turn be related to improvements in neurological performance.

Arctigenin Increases Hemeoxygenase-1 Gene Expression by Modulating PI3K/AKT Signaling Pathway in Rat Primary Astrocytes

  • Jeong, Yeon-Hui;Park, Jin-Sun;Kim, Dong-Hyun;Kim, Hee-Sun
    • Biomolecules & Therapeutics
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    • v.22 no.6
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    • pp.497-502
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    • 2014
  • In the present study, we found that the natural compound arctigenin inhibited hydrogen peroxide-induced reactive oxygen species (ROS) production in rat primary astrocytes. Since hemeoxygenase-1 (HO-1) plays a critical role as an antioxidant defense factor in the brain, we examined the effect of arctigenin on HO-1 expression in rat primary astrocytes. We found that arctigenin increased HO-1 mRNA and protein levels. Arctigenin also increases the nuclear translocation and DNA binding of Nrf2/c-Jun to the antioxidant response element (ARE) on HO-1 promoter. In addition, arctigenin increased ARE-mediated transcriptional activities in rat primary astrocytes. Further mechanistic studies revealed that arctigenin increased the phosphorylation of AKT, a downstream substrate of phosphatidylinositol 3-kinase (PI3K). Treatment of cells with a PI3K-specific inhibitor, LY294002, suppressed the HO-1 expression, Nrf2 DNA binding and ARE-mediated transcriptional activities in arctigenin-treated astrocyte cells. The results collectively suggest that PI3K/AKT signaling pathway is at least partly involved in HO-1 expression by arctigenin via modulation of Nrf2/ARE axis in rat primary astrocytes.

Studies on the anti-inflammatory action of Chilbokyeum extract in central nervous system (중추신경계(中樞神經系)에서 칠복음(七福飮)의 항염증작용(抗炎症作用)에 관한 연구(硏究))

  • Min Sang-Jun;Lee Sung-Ryull;Kang Hyung-Won;Lyu Yeoung-Su;Jeon Chang-Hwan
    • Journal of Oriental Neuropsychiatry
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    • v.12 no.2
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    • pp.173-183
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    • 2001
  • Substance P can stimulate secretion of tumor necrosis $factor-\;{\alpha}\;(TNF-\;{\alpha}\;)$ from astrocytes stimulated with lipopolysaccharide (LPS). Here I report that Chilbogeum can modulate cytokines secretion from primary cultures of rat astrocytes. Chilbogeum $(10\;{\mu}g/ml)$ significantly inhibited the $TNF-\;{\alpha}$ secretion by astrocytes stimulated with LPS and Substance P. Interleukin-1 (IL-1) has been shown to elevate $TNF-\;{\alpha}$ secretion from LPS-stimulated astrocytes while having no effect on astrocytes in the absence of LPS. Treatment of Chilbogeum $(10,\;100\;{\mu}g/ml)$ to astrocytes stimulated with both LPS and Substance P decreased IL-1 secretion significantly. The secretion of $TNF-\;{\alpha}$ by LPS and Substance P in astrocytes was progressively inhibited with increasing amount of IL-1 neutralizing antibody. Upon stimulation from various agents, these cells adopt a reactive phenotype, a morphological hallmark in Alzheimer's disease (AD) pathology, during which they themselves may produce still more inflammatory cytokines. Chilbogeum $(10,\;100\;{\mu}g/ml)$ significantly inhibited the $TNF-\;{\alpha}$ secretion by CCF-STTG1 astrocytoma cells stimulated with $A\;{\beta}$ and IL-1. These results suggest that Chilbogeum may inhibit $TNF-\;{\alpha}$ secretion by inhibiting IL-1 secretion and that Chilbogeum has an antiinflammatory activity in AD brain.

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Mannitol induces selective astroglial death in the CA1 region of the rat hippocampus following status epilepticus

  • Ko, Ah-Reum;Kang, Tae-Cheon
    • BMB Reports
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    • v.48 no.9
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    • pp.507-512
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    • 2015
  • In the present study, we addressed the question of whether treatment with mannitol, an osmotic diuretic, affects astrogliovascular responses to status epilepticus (SE). In saline-treated animals, astrocytes exhibited reactive astrogliosis in the CA1-3 regions 2-4 days after SE. In the mannitol-treated animals, a large astroglial empty zone was observed in the CA1 region 2 days after SE. This astroglial loss was unrelated to vasogenic edema formation. There was no difference in SE-induced neuronal loss between saline- and mannitol-treated animals. Furthermore, mannitol treatment did not affect astroglial loss and vasogenic edema formation in the dentate gyrus and the piriform cortex. These findings suggest that mannitol treatment induces selective astroglial loss in the CA1 region independent of vasogenic edema formation following SE. These findings support the hypothesis that the susceptibility of astrocytes to SE is most likely due to the distinctive heterogeneity of astrocytes independent of hemodynamics. [BMB Reports 2015; 48(9): 507-512]

Protein-protein interaction between caveolin-1 and SHP-2 is dependent on the N-SH2 domain of SHP-2

  • Park, Hyunju;Ahn, Keun Jae;Kang, Jihee Lee;Choi, Youn-Hee
    • BMB Reports
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    • v.48 no.3
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    • pp.184-189
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    • 2015
  • Src homology 2-containing protein tyrosine phosphatase 2 (SHP-2) is known to protect neurons from neurodegeneration during ischemia/reperfusion injury. We recently reported that ROS-mediated oxidative stress promotes phosphorylation of endogenous SHP-2 in astrocytes and complex formation between caveolin-1 and SHP-2 in response to oxidative stress. To examine the region of SHP-2 participating in complex formation with caveolin-1, we generated three deletion mutant constructs and six point mutation constructs of SHP-2. Compared with wild-type SHP-2, binding of the N-SH2 domain deletion mutant of SHP-2 to p-caveolin-1 was reduced greatly, using flow cytometric competitive binding assays and surface plasmon resonance (SPR). Moreover, deletion of the N-SH2 domain of SHP-2 affected $H_2O_2$-mediated ERK phosphorylation and Src phosphorylation at Tyr 419 in primary astrocytes, suggesting that N-SH2 domain of SHP-2 is responsible for the binding of caveolin-1 and contributes to the regulation of Src phosphorylation and activation following ROS-induced oxidative stress in brain astrocytes.