• Title/Summary/Keyword: primary glial cells

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Glial Fibrillary Acidic Protein Splice Variants in Hepatic Stellate Cells - Expression and Regulation

  • Lim, Michelle Chin Chia;Maubach, Gunter;Zhuo, Lang
    • Molecules and Cells
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    • v.25 no.3
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    • pp.376-384
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    • 2008
  • The glial fibrillary acidic protein (GFAP) is traditionally used as a marker for astrocytes of the brain, and more recently for the hepatic stellate cells (HSCs) of the liver. Several GFAP splice variants have been previously reported in the astrocytes of the CNS and in the non-myelinating Schwann cells of the PNS. In this study, we investigate whether GFAP splice variants are present in the HSCs and their expression as a function of HSCs activation. Furthermore, the regulation of these transcripts upon treatment with interferon gamma ($IFN-{\gamma}$) will be explored. Using semi-quan-titative RT-PCR and real-time PCR, we examine the expression and regulation of GFAP splice variants in HSCs as well as their respective half-life. We discover that most of the GFAP splice variants ($GFAP{\alpha}$, ${\beta}$, ${\delta}$, ${\varepsilon}$ and $\kappa$) found in the neural system are also expressed in quiescent and culture-activated primary HSCs. Interestingly, $GFAP{\alpha}$ is the predominant form in quiescent and culture-activated primary HSCs, while $GFAP{\beta}$, predominates in the SV40-immortalized activated HSC-T6. $GFAP{\delta}$, ${\varepsilon}$ and ${\kappa}$ have similar half-lives of 10 hours, while $GFAP{\beta}$ has a half-life of 17 hours. Treatment of HSC-T6 with $IFN-{\gamma}$ results in a significant 1.29-fold up-regulation of $GFAP{\alpha}$ whereas the level of the other transcripts remains unchanged. In summary, $GFAP{\alpha}$, ${\beta}$, ${\delta}$, ${\varepsilon}$ and $\kappa$ are present in HSCs. They are differentially regulated on the transcription level, implying a role of the 5' and 3' untranslated regions.

Flavonoids Modulate the Proliferation of Neospora caninum in Glial Cell Primary Cultures

  • Matos, Rosan Barbosa De;Braga-de-Souza, Suzana;Pitanga, Bruno Pena Seara;Silva, Victor Diogenes Amaral Da;Jesus, Erica Etelvina Viana De;Pinheiro, Alexandre Morales;Costa, Maria De Fatima Dias;El-Bacha, Ramon Dos Santos;Ribeiro, Catia Suse De Oliveira;Costa, Silvia Lima
    • Parasites, Hosts and Diseases
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    • v.52 no.6
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    • pp.613-619
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    • 2014
  • Neospora caninum (Apicomplexa; Sarcocystidae) is a protozoan that causes abortion in cattle, horses, sheep, and dogs as well as neurological and dermatological diseases in dogs. In the central nervous system of dogs infected with N. caninum, cysts were detected that exhibited gliosis and meningitis. Flavonoids are polyphenolic compounds that exhibit antibacterial, antiparasitic, antifungal, and antiviral properties. In this study, we investigated the effects of flavonoids in a well-established in vitro model of N. caninum infection in glial cell cultures. Glial cells were treated individually with 10 different flavonoids, and a subset of cultures was also infected with the NC-1 strain of N. caninum. All of the flavonoids tested induced an increase in the metabolism of glial cells and many of them increased nitrite levels in cultures infected with NC-1 compared to controls and uninfected cultures. Among the flavonoids tested, 3',4'-dihydroxyflavone, 3',4',5,7-tetrahydroxyflavone (luteolin), and 3,3',4',5,6-pentahydroxyflavone (quercetin), also inhibited parasitophorous vacuole formation. Taken together, our findings show that flavonoids modulate glial cell responses, increase NO secretion, and interfere with N. caninum infection and proliferation.

A Case of Cerebral Metastasis from Malignant Fibrous Histiocytoma (뇌로 전이한 악성 섬유성 조직구종 1례)

  • Kang, Kwan-Soo;Lee, Jung-Il;Suh, Yeon Lim
    • Journal of Korean Neurosurgical Society
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    • v.30 no.11
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    • pp.1340-1344
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    • 2001
  • This is a rare case of cerebral metastasis from malignant fibrous histiocytoma(MFH) of the soft tissue. A 62-year-old man underwent craniotomy for resection of multiple intracerebral masses under the impression of metastatic brain tumor with unknown primary site. Preoperative investigation failed to detect any extracranial lesion. At six months after the operation and whole brain radiotherapy, right shoulder mass was detected to grow and excised. Specimen from the brain and shoulder lesions revealed identical pathological findings of malignant fibrous histiocytoma except existence of glial fibrillary acidic protein(GFAP)-positive cells only in brain lesions. Palliative radiotherapy was performed for subsequently developing metastatic lesions in skeletal system. At twelve months after initial diagnosis recurrent lesion at right shoulder was detected and chemotherapy is given. This case is unique because metastatic brain lesion from MFH is rare and also cerebral metastasis as an initial manifestaion of MFH has not been reported before. Another important finding is that there was expression of GFAP only in brain lesions but not in extracranial primary site lesion. Although the presence of GFAP-positive cells is thought as one of characteristic histological findings of primary intracrainal MFH, our observation supports the hypothesis that GFAP-positive cells in primary intracranial MFH may be nonneoplastic astrocytes secondarily involved by MFH.

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Effect of glucose level on chemical hypoxia- and hydrogen peroxide-induced chemokine expression in human glioblastoma cell lines

  • Jung, Yieun;Ahn, So-Hee;Park, Sang Hui;Choi, Youn-Hee
    • The Korean Journal of Physiology and Pharmacology
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    • v.21 no.5
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    • pp.509-518
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    • 2017
  • Glioblastoma multiforme (GBM) is the most common primary intracranial tumor in adults and has poor prognosis. The GBM-specific tumor microenvironment (TME) plays a crucial role in tumor progression, immune escape, local invasion, and metastasis of GBM. Here, we demonstrate that hypoxia, reactive oxygen species (ROS), and differential concentration of glucose influence the expression of cytokines and chemokines, such as IL-6, IL-8, and IP-10, in human glial cell lines. Treatment with cobalt chloride ($CoCl_2$) and hydrogen peroxide ($H_2O_2$) significantly increased the expression levels of IL-6, IL-8, and IP-10 in a dose-dependent manner in CRT-MG and U251-MG astroglioma cells, but not in microglia cells. However, we found strikingly different patterns of expression of cytokines and chemokines between $H_2O_2$-treated CRT-MG cells cultured in low- and high-glucose medium. These results suggest that astroglioma and microglia cells exhibit distinct patterns of cytokine and chemokine expression in response to $CoCl_2$ and $H_2O_2$ treatment, and different concentrations of glucose influence this expression under either hypoxic or oxidant-enriched conditions.

A Case of Astroblastoma

  • Han, Young-Min;Kim, Jong-Tae;Chung, Dong-Sup;Park, Young-Sup
    • Journal of Korean Neurosurgical Society
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    • v.40 no.5
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    • pp.373-376
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    • 2006
  • An astroblastoma is a rare primary glial tumor occurring preferentially in young adults. It is characterized by a perivascular arrangement of the tumor cells forming perivascular pseudorosettes mimicking ependymomas. The histogenesis of astroblastoma is unclear, despite a number of studies to determine its possible cellular origin. We have experienced a case of astroblastoma located at the temporal lobe. It presented as a large, well-circumscribed, and highly enhanced mass lesion on magnetic resonance images[MRI]. The tumor was well demarcated and did not infiltrate the brain, which made complete removal possible. Here, we report and discuss the characteristic histological and radiological features of this case.

Neurotoxicity Assessment of Methamphetamine and Cadmium Using Cultured Neuronal Cells of Long-Evans Rats (신경세포 배양법을 이용한 methamphetamine과 cadmium의 신경독성 평가)

  • Cho, Dae-Hyun;Kim, Jun-Gyon;Jeong, Yong;Lee, Bong-Hun;Kim, Eun-Youb;Kim, Jeong-Goo;Cho, Tai-Soon;Kim, Jin-Suk;Moon, Hwa-Hwey
    • Toxicological Research
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    • v.12 no.1
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    • pp.69-79
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    • 1996
  • Primary culture of cerebellar neuronal cells derived from 8-day old Long-Evans rats was used. Pure granule cells, astrocytes or mixed cells culture systems were prepared. These cells were differentiated and developed synaptic connections. And the astrocytes were identified by immunostaining with glial fibrillary acidic protein (GFAP). Methamphetamine (MAP), which acts on dopaminergic system and cadmium (Cd), a toxic heavy metal, were applied and biochemical assays and electrophysiological studies were performed. $LC_50$ values estimated by MTT assay of MAP and Cd were 3 mM and 2$\mu M$ respectively. Cells were treated with 1 mM or 2 mM MAP and 1$\mu M$ $CdCl_2$ for 48 hour, and the incubation media were analyzed for the content of released LDH. MAP (2 mM) and Cd significantly increased the LDH release. Cell viability was decreased in both groups and some cytopathological changes like cell swelling or vacuolization were seen. The cerebellar granule cells were used for measuring membrane currents using whole-cell clamp technique. Sodium and potassium currents were not affected by MAP neither Cd, but calcium current was significantly reduced by Cd but not affected by MAP. Therefore, in vitro neurotoxicity test system using neuronaI cells and astrocytes cultures were established and can be used in screening of potential neurotoxic chemicals.

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Malignant Retroperitoneal Paraganglioma in a Siberian Tiger (시베리아 호랑이에서 발생한 악성후복막부신경절종 증례)

  • Shin, Sang-Kyung;Park, Byung-Min;Youm, So-Young;Kim, Tae-Wang;Na, Ki-Jeong;Ahn, Byeong-Woo
    • Journal of Veterinary Clinics
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    • v.28 no.5
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    • pp.522-525
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    • 2011
  • A 14 year old female Siberian tiger presented for postmortem examination. A large mass attached to sublumbar area was found to be circumscribing aorta with metastases to mesenteric lymph nodes, uterus, kidney, adrenal gland, lung and thymus. The tumor cells were arranged in clusters or nests separated by well-developed fibrovascular stroma. The individual cells were plump and polygonal with granular eosinophilic cytoplasms and had distinct cell borders. The tumor cells were positive for synaptophysin, chromogranin A and neuron-specific enolase, and negative for cytokeratins, S100 and glial fibrillary acidic protein. The primary tumor was diagnosed as a malignant retroperitoneal paraganglioma.

Glioblastoma in a Pekingese (페키니즈견의 아교모세포종 증례)

  • Cho, Hyun-kee;Yoo, Dae-Young;Kang, Joo-yeon;Lee, Kwon-Young;Hwang, In-Koo;Choi, Jung-Hoon;Chung, Jin-Young
    • Journal of Veterinary Clinics
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    • v.32 no.6
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    • pp.544-547
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    • 2015
  • An 11-year-old, intact male Pekingese was brought to the Veterinary Teaching Hospital of Kangwon National University with a 10-day history of seizures. Fifteen days before coming to Kangwon National University, the dog had visited a local animal hospital for lameness, and non-steroidal anti-inflammatory drugs were prescribed to treat this symptom. However, 10 days before coming to our hospital, the dog experienced generalized seizures. Two days before his arrival, generalized ataxia and mental dullness also occurred. Our examinations revealed no remarkable findings on a routine blood test or X-ray. However, the neurological examinations confirmed mental dullness, generalized ataxia, and a lack of menace response and pupillary light reflexes. Nine hours later, dyspnea occurred, and 12 hours after that, the patient was euthanized per the client's request. A necropsy of transverse sections confirmed the presence of a prominent midline shift due to extended tumor growth. On histopathological analyses, pseudopalisading necrosis of the glial cells and microvascular proliferation were observed. In immunohistochemical analysis, glial fibrillary acidic protein, proliferating cell nuclear antigens, and ionized calcium binding adaptor molecule 1 immunoreactive cells were observed in the tumor area. Based on the results, the tumor was confirmed to be a glioblastoma. Primary intracranial tumors are rare in the veterinary field. This case report describes the clinical and histopathological findings of glioblastoma in a Pekingese.

Anti-parkinsonian effect of Cyperi Rhizoma via inhibition of neuroinflammatory action (향부자(香附子)의 염증 억제 작용을 통한 항파킨슨 효과)

  • Kim, Hyo Geun;Sim, Yeomoon;Oh, Myung Sook
    • The Korea Journal of Herbology
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    • v.28 no.5
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    • pp.21-28
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    • 2013
  • Objectives : The aim of this study was to investigate the neuroprotective effects and mechanisms of Cyperi Rhizoma extracts (CRE) using in vitro and in vivo models of Parkinson's disease (PD). Methods : We evaluated the neuroprotective effect of CRE against 1-methyl-4-phenylpyridinium (MPP+) toxicity using tyrosine hydroxylase immunohistochemistry (IHC) in primary rat mesencephalic dopaminergic neurons. In addition, the effect of CRE was evaluated in mice PD model induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). For evaluations, C57bl/6 mice were orally treated with CRE 50 mg/kg for 5 days and were injected intraperitoneally with MPTP (20 mg/kg) at 2 h intervals on the last day. To identify the CRE affects on MPTP-induced neuronal loss of dopaminergic neurons in substantia nigra pars compacta (SNpc) and striatum of mice, the behavioral tests and IHC analysis were carried out. Also, we conducted nitric oxide (NO) and tumor necrosis factor-alpha (TNF-${\alpha}$) assay in dopaminergic neurons and IHC using glial markers in SNpc of mice to assess the anti-inflammation effects. Results : In primary mesencephalic culture system, CRE protected dopaminergic cells against $10{\mu}M$ MPP+-induced toxicity at 0.2 and $1.0{\mu}g/mL$. In the behavior tests, CRE treated group showed improved motor deteriorations than those in the MPTP only treated group. CRE significantly protected striatal dopaminergic damage from MPTP-induced neurotoxicity in mice. Moreover, CRE inhibited productions of NO and TNF-${\alpha}$ in dopaminergic culture system and activation of astrocyte and microglia in SNpc of the mice. Conclusion : We concluded that CRE shows anti-parkinsonian effect by protecting dopaminergic neurons against MPP+/MPTP toxicities through anti-inflammatory actions.

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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