• The Journal of Korean Medicine
• /
• v.29 no.1
• /
• pp.167-178
• /
• 2008

Object : Gliosis becomes a physical and mechanical barrier to axonal regeneration. Reactive gliosis induced by hypoxic brain injury is involved with up-regulation of CD81 and GFAP. The current study was to examine the effect of the Angelica Sinens on CD81 and GFAP regulation after hypoxic brain injury in the astrocyte. Methods : MTT assay was performed to examine cell viability, and cell based ELISA, western blot and PCR were used to detect the expression of CD81 and GFAP. Results : The following results were obtained: 1. Using ELISA, western blot and PCR from the astrocyte after hypoxic injury, CD81 and GFAP expression was seen to have increased. 2. After the administration of Angelica Sinens extract to astrocyte following hypoxic injury, CD81 and GFAP expression was down regulated significantly. The water extract of Angelica Sinens prevented cell destruction by hypoxic induced with $CoCl_2$. Conclusion : These results indicate that Angelica Sinens could suppress reactive gliosis, which disturbs astrocyte regeneration after hypoxic brain injury by controlling the expression of CD81 and GFAP.

• The Korean Journal of Pain
• /
• v.11 no.2
• /
• pp.182-193
• /
• 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.

• Korean Journal of Exercise Nutrition
• /
• v.16 no.2
• /
• pp.73-84
• /
• 2012

Thyroid hormones are important for the development of the brain including the cerebellum. In the present study, we investigated the effect of treadmill exercise on the survival of Purkinje neurons and the activation of astrocytes in the cerebellar vermis of hypothyroidism-induced rat pups. On the day of perinatal 14, pregnant rats were divided into two groups (n = 5 in each group): the pregnant control group and the pregnantmethimazole (MMI)-treated group. For the induction of hypothyroidism in the rat pups, MMI was added to the drinking water (0.02% wt/vol), from the day of perinatal 14 to postnatal 49. After delivery, male rat pups born from the pregnant control group were assigned to the control group. Male rat pups born from the MMI-treated group were divided into the hypothyroidism-induction group, the hypothyroidism-induction with treadmill exercise group, and the hypothyroidism-induction with thyroxine (T4) treatment group (n = 10 in each group). The rat pups in the exercise group were forced to run on a treadmill for 30 min once a day for 4 weeks, starting on postnatal day 22. In the hypothyroidism-induced rat pups, motor coordination was reduced and Purkinje cell death and reactive astrocytes in the cerebellar vermis were increased. Treadmill exercise enhanced motor coordination, increased the survival of Purkinje neurons, down-regulated reactive astrocytes, and enhanced brain-derived neurotrophic factor (BDNF) and receptor tyrosine kinase B (TrkB) expressions in the hypothyroidism-induced rat pups. These results suggest that treadmill exercise has beneficial effects in terms of protecting against thyroid dysfunction by increasing T3 and T4 and the related protein, BDNF, as well as TrkB, inhibition on astrocyte activation and the reduction of Purkinje cell loss regarding the cerebellum in hypothyroidism rat pups.

• The Journal of Korean Medicine
• /
• v.39 no.4
• /
• pp.1-15
• /
• 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.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.21 no.1
• /
• pp.145-152
• /
• 2007

Following CNS injury, inhibitory influences at the site of axonal damage occur. Glial cells become reactive and form a glial scar, gliosis. Astrocyte-rich gliosis relates with up-regulation of GFAP and CD81, and eventually becomes physical and mechanical barrier to axonal regeneration. It is postulated that the astrocytic reaction is absent, regeneration of axons can occur. And it was reported that treatment with anti CD81 antibodies enhanced functional recovery in the rat with spinal cord injury. So in this current study, the author investigated the effect of the water extract of Persicae Semen on the regulation of GFAP and CD81 that increase when gliosis occurs. Persicae Semen decreased the expression of GFAP and CD81 in astrocyte cell by ELISA method. Persicae Semen decreased the RNA expression of CD81 and GFAP. The proteins that separate in whole cell were analaysed by western blot, and the expression of GFAP and CD81 was decreased. In vivo, rats brains were peformed cortical stab wound, the water extracts of Persicae Semen were injected for 7 days, 30 days. As a result, GFAP and CD81 expression were decreased in immunohistochemistry. These findings demonstrate that Persicae Semen decreases GFAP and CD81 expression. Accordingly, Persicae Semen could be a candidate for promotion of axon regeneration after CNS injury.

• Biomolecules & Therapeutics
• /
• v.22 no.6
• /
• pp.497-502
• /
• 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.

• The Korean Journal of Physiology and Pharmacology
• /
• v.7 no.6
• /
• pp.307-310
• /
• 2003

Kainic acid (KA) is a structural analogue of glutamate that interacts with specific presynaptic and postsynaptic receptors to potentiate the release and excitatory actions of glutamate. Systemic or intracerebroventricular (i.c.v.) administration of KA to experimental animals elicits multifocal seizures with a predominantly limbic localization, and results in neuronal death of cornu ammonia 1 (CA1), reactive gliosis and biochemical changes in the hippocampus and other limbic structures. Several lines of evidence suggest that reactive oxygen species (ROS) play a pivotal role in the pathogenesis of excitotoxic death by KA. Curcumin has been known to possess anti-oxidative and anti-inflammatory activities. In this study, the effects of curcumin on KA induced hippocampal cell death, reactive gliosis and biochemical changes in reactive glia were investigated by immunohistochemical methods. Our data demonstrated that curcumin attenuated KA-induced astroglial and microglial activation although it did not protect KA-induced hippocampal cell death.

• The Korean Journal of Physiology and Pharmacology
• /
• v.1 no.3
• /
• pp.285-296
• /
• 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.

• Journal of Pharmacopuncture
• /
• v.12 no.1
• /
• pp.67-76
• /
• 2009

Objective : Following central nervous system(CNS) injury, inhibitory influences at the site of axonal damage occur. Glial cells become reactive and form a glial scar, gliosis. Also myelin debris such as MAG inhibits axonal regeneration. Astrocyte-rich gliosis relates with up-regulation of GFAP and CD81, and eventually becomes physical and mechanical barrier to axonal regeneration. MAG is one of several endogenous axon regeneration inhibitors that limit recovery from CNS injury and disease. It was reported that molecules that block such inhibitors enhanced axon regeneration and functional recovery. Recently it was reported that treatment with anti-CD81 antibodies enhanced functional recovery in the rat with spinal cord injury. So in this current study, the author investigated the effect of the water extract of Uncariae Ramulus et Uncus on the regulation of CD81, GFAP and MAG that increase when gliosis occurs. Methods : MTT assay was performed to examine cell viability, and cell-based ELISA, western blot and PCR were used to detect the expression of CD81, GFAP and MAG. Then also immunohistochemistry was performed to confirm in vivo. Results : Water extract of Uncariae Ramulus et Uncus showed relatively high cell viability at the concentration of 0.05%, 0.1% and 0.5%. The expression of CD81, GFAP and MAG in astrocytes was decreased after the administration of Uncariae Ramulus et Uncus water extract. These results was confirmed in the brain sections following cortical stab injury by immunohistochemistry. Conclusion : The authors observed that Uncariae Ramulus et Uncus significantly down-regulates the expression of CD81, GFAP and MAG. These results suggest that Uncariae Ramulus et Uncus can be a candidate to regenerate CNS injury.

• The Journal of Internal Korean Medicine
• /
• v.30 no.1
• /
• pp.24-35
• /
• 2009

Object : In conditions of brain infarction, irreversible axon damage occurs in the central nerve system (CNS), because gliosis makes physical and mechanical barriers. If gliosis formation could be suppressed, irreversible axon damage would be reduced. This could mean that an injured CNS could be regenerated. CD81 and GFAP have close relationships to gliosis. The increase in glial cells at CNS injury gives rise to the expression of CD81 and GFAP. CD81 was postulated to play a central role in the process of CNS scar formation. Method : In this study, the author investigated the effect of the water extract of the Moutan Radicis Cortex on regulation of CD81 and GFAP expression in injured CNS cells. MTT assay was used to examine cell viability, while RT-PCR and ELISA methods were carried out to measure the expression of CD81 and GFAP in the astrocyte. Results : We observed that water extract of the Moutan Radicis Cortex increased cell viability under hypoxia induced by $CoCl_2$ and suppressed the expression of CD81 and GFAP up-regulated by hypoxia. Conclusion : These results suggest that the Moutan Redicis Cortex could promote neural regeneration as a consequence of protecting CNS cells from hypoxia and suppressing the reactive gliosis following CNS injury.