• International Journal of Oral Biology
• /
• v.40 no.2
• /
• pp.71-77
• /
• 2015

The activation of glial cells in the spinal cord has been contribute to the initiation and maintenance of pain facilitation induced by peripheral inflammation and nerve injury. The present study investigated effects of botulinum toxin type A (BoNT-A), injected subcutaneously or intracisternally, on the expression of microglia and astrocytes in rats. Complete Freund's Adjuvant (CFA)-induced inflammation was employed as an orofacial chronic inflammatory pain model. A subcutaneous injection of $40{\mu}L$ CFA into the vibrissa pad was performed under 3% isoflurane anesthesia in SD rats. Immunohistochemical analysis for changes in Iba1 (a microglia marker) and GFAP (an astrocyte marker), were performed 5 days after CFA injection. Subcutaneous injection of CFA produced increases in Iba1 and GFAP expression, in the ipsilateral superficial lamia I and II in the medullary dorsal horn of rats. Subcutaneous treatment with BoNT-A attenuated the up-regulation of Iba1 and GFAP expressions induced by CFA injection. Moreover, intracisternal injection of BoNT-A also attenuated the up-regulated Iba1 and GFAP expressions. These results suggest that the anti-nociceptive action of BoNT-A is mediated by modulation activation of glial cells, including microglia and astrocyte.

• Preventive Nutrition and Food Science
• /
• v.19 no.3
• /
• pp.145-155
• /
• 2014

Vascular dementia is characterized by white matter lesions involving the demyelination and activation of astrocytes and microglia. In a previous study, we showed that the supernatant of a laboratory-scale, hot water extract of ground whole wheat (TALE) attenuated white matter injury and astrocytic activation in a rat model of bilateral common carotid artery occlusion (BCCAO). In the present study, we made several modifications to the hot water extraction process to remove starch and enable large-scale production. We used wheat bran (WB), which contains less starch, instead of ground whole wheat. In addition, we removed starch granules with a decanter before hot water extraction. The final product, wheat bran extract (WBE), contained 2.42% arabinose, a surrogate marker of arabinoxylan, which is an active constituent of WBE. Supplementation of the rat model of BCCAO with WBE (400 mg/kg/day) for 33 days attenuated white matter injury, which was assessed by Luxol Fast Blue staining, in the corpus callosum (cc) and optic tract (opt) regions. Attenuation of white matter injury in the opt region was accompanied by improvement of the pupillary light reflex. Immunochemical staining revealed that supplementation with WBE reduced astrocytic activation in the cc and opt regions and reduced microglial activation in the opt region. These findings indicate that supplementation with WBE is effective at attenuating white matter injury accompanied by the inhibition of astrocytic and microglial activation. Therefore, extracts from WB, a cheap by-product of wheat milling, can be developed as a nutraceutical to prevent vascular dementia, a disease for which there is no approved pharmaceutical treatment.

• Journal of Life Science
• /
• v.17 no.5 s.85
• /
• pp.706-713
• /
• 2007

Scutellaria baicalensis GEORGI(SB) is used in oriental medicine for the treatment of incipient strokes. Although it has been reported that SB is neuroprotective in a hypoxia model, its mechanism is poorly understood. Here, we investigated the effect of SB on the modulation of heme oxygenase-1(HO-1), which has important biological roles in regulating mitochondrial heme protein turnover and in protecting against conditions such as hypoxia, neurodegenerative diseases, or sepsis. Rat cerebrocortical day In vitro(DIV)12 cells were grown in neurobasal medium. On DIV12 cells were treated with SB($20{\mu}g/ml$) and given a hypoxic shock ($2%\;O_2/5%\;CO_2,\;3\;hr$) on DIV14. In situ hybridization results revealed that SB upregulated HO-1 mRNA in neuronal dendrites in both normoxia and hypoxia(38.5% and 59.2%, respectively). At the protein level, SB upregulated HO-1 in the neuronal soma in both normoxia and hypoxia(22.4% and 15.7%, respectively). Interestingly, most significant increase was associated with astrocytes, which increased HO-1 protein by 77.5% compared to SB-untreated culture. These results indicate that SB upregulates both neuronal and glial HO-1 expression, which contributes to the neuroprotection efficacy in hypoxia).

• Korean Journal of Veterinary Research
• /
• v.46 no.3
• /
• pp.177-184
• /
• 2006

Experimental autoimmune encephalomyelitis (EAE) is a disease model of multiple sclerosis (MS) that is characterized by remittance and relapse of the disease and autoimmune and demyelinating lesions in the central nervous system (CNS). Autoimmune inflammation is maintained by secretion of a large number of protein. Previous studies have suggested that transcripts encoding osteopontin (OPN) are frequently detected in the mRNA population of MS plaques. To elucidate the functional role of OPN in initiation and development of EAE, we examined the expression and localization of OPN in the spinal cord during acute EAE. We demonstrated that OPN significantly increased at the early stage of EAE and slightly declined thereafter by western blot analysis. An immunohistochemical study revealed that OPN was constitutively expressed in some glial cells (microglia, astrocytes) of white matter and neurons in the CNS of control rats. OPN expression was shown to be increased in the same cells at the early and peak stage of EAE. To identity cells expressing OPN by double-immunofluorescence labeling, we labeled rat spinal cord sections for OPN with a monoclonal OPN antibody and with mAbs for astrocyte (GFAP), microglia/macrophage (OX42)-specific markers. The major cell types of OPN-expressing cells were activated astrocytes and microglia in the adjacent inflammatory lesions. Interestingly, OPN was mainly expressed in the end feet of astrocytes around vascular cell adhesion molecule-1 (VCAM-1) expressing endothelial cells of CNS blood vessel. These findings suggest that increased levels of OPN in activated glial cell may play an important role in the recruitment of inflammatory cells into the CNS parenchyma during EAE.

• The Korea Journal of Herbology
• /
• v.35 no.4
• /
• pp.9-16
• /
• 2020

Objective : To identify the effects of the water extract of Liriope platyphylla tuber (Liriopis tuber, LT) on the activation of astocytes, we investigated the regulatory effects of LT extract on H₂O₂-induced oxidative damage in C6 rat astrocytes. Methods : LT extract was extracted with boiling water. C6 cell line were treated with LT extract at 1, 2, and 3 mg/㎖ or without for 30 min and then stimulated with H₂O₂ at 5 ㎛ for 24 hr. The cell viability was measured by MTT assay. The expression of glial fibrillary acidic protein (GFAP), signal transducer and activator of transcription 3 (STAT3), phospho-STAT3 (pSTAT3), cyclooxygenase (COX-2), Nuclear factor-κB (NF-κB), superoxide dismutase 2 (SOD2), heme oxygenase-1 (HO-1), catalase, Akt, phospho-Akt (p-Akt) phosphoinositide 3-kinases (PI3K), and protein kinase C alpha (PKCα) proteins were determined by Western blot, respectively. GFAP expression was also observed with immunocytochemistry under a fluorescence microscope. Results : LT extract induced cell proliferation in H₂O₂-stimulated C6 cells. LT extract significantly inhibited the expression of GFAP, NF-κB and COX-2 and increased the expression of HO-1 and the phosphorylation of STAT3 in H₂O₂-stimulated C6 cells. LT extract also significantly increased the phosphorylation of Akt and decreased the expression of PKCα in a dose-dependent manner in H₂O₂-stimulated C6 cells. Conclusions : LT extract can regulate H₂O₂-induced activation of astrocytes through inhibiting the expression of NF-κB, COX-2 and regulating Akt / HO-1, STAT3 or PKCα signaling pathway.

• Toxicological Research
• /
• v.11 no.2
• /
• pp.315-327
• /
• 1995

Diverse neurotoxic insults result in proliferation and hypertrophy of astrocytes, a subtype of glia in central nervous system. The hallmark of this response, often terms "reactive gliosis", is the enhanced expression of the major intermediate filament protein of castrocytes, glial fibrillary acidic protein (GFAP). These changes in the astrocytes suggest that GFAP may be a useful biochemical indicator of neurotoxicity. To investigate this possibility, we administered intra-peritoneally prototype nerotoxicants, metharnphetamine (MAP, 5 mg/kg), cocaine (30 mg/kg), N-buthyl benzenesulfonamide (NBBS, 300 mg/kg) and trimethytin (TMT, 8 mg/kg) to Wistar Rats and then assessed the effects of these agents on content of GFAP, which were determined by Sandwish ELISA and evaluated with neurotoxic symptoms, and quantitative changes of imrnunoreactivity of GFAP by light microscopic image analysis in specific regions. We found that assay of GFAP revealed time- and region-dependant patterns of neurotoxicity. The GFAP immunoreactivity of rat brain was increased in substantia nigra and hippocampus by MAP, NBBS and TMT; in roedial septal nucleus and nucleus accurnbens, it was also increased by RrBBS. Sandwich ELISA showed that GFAP levels of cerebrum in all groups on days 3 and 7 and that of brainstem(including cerebellum) in MAP, NBBS groups on day 1 and 3 were increased. A review of the background, design and results of these experiments are presented in this paper. Our findings indicate that GFAP is a sensitive and specific biomarker of neurotoxicity.otoxicity.

• Toxicological Research
• /
• v.32 no.1
• /
• pp.73-80
• /
• 2016

Chronic exposure to cadmium (Cd) is known to adversely affect renal function. Our previous studies indicated that Cd induces p53-dependent apoptosis by inhibiting gene expression of the ubiquitin-conjugating enzyme (Ube) 2d family in both human and rat proximal tubular cells. In this study, the effects of Cd on protein expression of p53 and apoptotic signals in the kidney and liver of mice exposed to Cd for 12 months were examined, as well as the effects of Cd on p53 protein levels and gene expression of the Ube2d family in various cell lines. Results showed that in the kidney of mice exposed to 300 ppm Cd for 12 months, there was overaccumulation of p53 proteins in addition to the induction of apoptosis, which was triggered specifically in the proximal tubules. Interestingly, the site of apoptosis was the same as that of p53 accumulation in the proximal tubules. In the liver of mice chronically exposed to Cd, gene expression of the Ube2d family tended to be slightly decreased, together with slight apoptosis without the accumulation of p53 protein. In rat small intestine epithelial (IEC-6) cells, Cd decreased not only the p53 protein level but also gene expression of Ube2d1, Ube2d2 and Ube2d4. In human brain microvascular endothelial cells (HBMECs), Cd did not suppress gene expression of the Ube2d family, but increased the p53 protein level. In human brain astrocytes (HBASTs), Cd only increased gene expression of UBE2D3. These results suggest that Cd-induced apoptosis through p53 protein is associated with renal toxicity but not hepatic toxicity, and the modification of p53 protein by Cd may vary depending on cell type.

• Korean Journal of Veterinary Research
• /
• v.40 no.3
• /
• pp.431-437
• /
• 2000

The aim of this study was to evaluate the involvement of CPP32 (caspase-3), one of the death-related enzymes, in the course of experimental autoimmune encephalomyelitis (EAE). EAE was induced in Lewis rats immunized with an emulsion of rat spinal cord homogenate with complete Freunds adjuvant supplemented with Mycobacterium tuberculosis (H37Ra, 5mg/ml). The expression of CPP32 in the spinal cords of rats with EAE was studied. In normal rat spinal cords, CPP32 is constitutively, but weakly, expressed in neurons and some neuroglial cells. In the EAE spinal cords, many inflammatory cells were positive for CPP 32, and the majority of CPP32(+) cells were identified as ED1(+) macrophages. During this stage of EAE, the number of CPP32(+) cells in brain cells, including neurons and astrocytes, increased, and these cells also had increased CPP32 immunoreactivity. CPP32 immunor eactivity was not always matched with apoptosis of inflammatory cells in EAE lesions. We speculate that CPP32, which is constitutlvely expressed in brain cells, increases in response to neuroimmunological stimulation in both brain neuronal cells and inflammatory cells. The functional role of CPP32 in neuroimmunological disorders is discussed.

• Journal of Embryo Transfer
• /
• v.32 no.2
• /
• pp.39-45
• /
• 2017

Mesenchymal stem cells (MSCs) have been considered an alternative source of neuronal lineage cells, which are difficult to isolate from brain and expand in vitro. Previous studies have reported that MSCs expressing Nestin ($Nestin^+$ MSCs), a neuronal stem/progenitor cell marker, exhibit increased transcriptional levels of neural development-related genes, indicating that $Nestin^+$ MSCs may exert potential with neurogenic differentiation. Accordingly, we investigated the effects of the presence of $Nestin^+$ MSCs in bone-marrow-derived primary cells (BMPCs) on enhanced neurogenic differentiation of BMPCs by identifying the presence of $Nestin^+$ MSCs in uncultured and cultured BMPCs. The percentage of $Nestin^+$ MSCs in BMPCs was measured per passage by double staining with Nestin and CD90, an MSC marker. The efficiency of neurogenic differentiation was compared among passages, revealing the highest and lowest yields of $Nestin^+$ MSCs. The presence of $Nestin^+$ MSCs was identified in BMPCs before in vitro culture, and the highest and lowest percentages of $Nestin^+$ MSCs in BMPCs was observed at the third (P3) and fifth passages (P5). Moreover, significantly the higher efficiency of differentiation into neurons, oligodendrocyte precursor cells and astrocytes was detected in BMPCs at P3, compared with P5. In conclusion, these results demonstrate that neurogenic differentiation can be enhanced by increasing the proportion of $Nestin^+$ MSCs in cultured BMPCs.

• Journal of Oriental Neuropsychiatry
• /
• v.20 no.1
• /
• pp.177-197
• /
• 2009

Objectives : Gliosis disturbs recovery of damaged astrocytes following central nervous system(CNS) injury. Gliosis relates to up-regulation of CD81 and GFAP. In glial cells at injured CNS, the expression of CD81 and GFAP is increased. It is possible that when the gliosis formation is suppressed, regeneration of oxons can occur. According to the recent study, the treatment with anti CD81 antibodies enhanced functional recovery in rats with spinal injury. So, the author studies the effect of water extract of Radix Ginseng on regulation of CD81 and GFAP with CNS injury. Methods : In the cell study, hypoxic damage was induced by CoC12. And according to Longa et al, cerebral ischemia was made by middle cerebral artery occlusion in the rat. Cross sections of rat brain were examined under microscope. MTT analysis was performed to examine cell viability, cell based ELISA, Western Blot and PCR were used to detect the expression of CD81 and GFAP. Results : The following results were obtained. 1. We found that CD81 and GFAP were decreased in hypoxic injured cells following Radix Ginseng administration. 2. We injected the extract of Radix Ginseng to the middle cerebral artery occlusion in rats, and the immunohistochemistry analysis showed that CDS1 and GFAP were decreased. Conclusions : These results show that the extract of Radix Ginseng could suppress the gliosis formation and prevent cell death, by controlling the expression of CDS1 and GFAP. Therefore, Radix Ginseng could be a useful to regenerate CNS injury.