• Title/Summary/Keyword: cell homogenates

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Inhibition of Oxidative Stress-induced and Excitotoxic Neuronal Cell Damage by Xuesaitong Ruanjiaonang (혈색통연교낭(血塞通軟膠囊)의 산화적 및 흥분성 신경세포독성 억제작용)

  • Cho Jungsook
    • YAKHAK HOEJI
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    • v.49 no.1
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    • pp.30-37
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    • 2005
  • Xuesaitong Ruanjiaonang (XR), a soft capsule containing Panax notoginseng saponins as main ingredients, is believed to remove extravasated blood and increase cerebral blood flow by improving blood circulation, and therefore, has been used in China to treat ischemic stroke or hemiplegia caused by cerebral thrombosis. To characterize pharmacological actions of XR, the present study evaluated its effects on neuronal cell damage induced by various oxidative insults or excitotoxic amino acids in primary cultured rat cortical cells. The neuronal cell viability was not affected by XR with the exposure for 2 h at the concentrations tested in this study ($10{\sim}1000\;{\mu}g/ml$). However, significant reduction of the cell viability was observed when the cultured cells were exposed to XR at $1000\;{\mu}g/ml$ for 24 h. XR was found to concentration-dependently inhibit the oxidative neuronal damage induced by $H_{2}O_2$, xanthine/xanthine oxidase or $Fe^{2+}$/ascorbic acid. In addition, it dramatically inhibited the excitotoxic damage induced by glutamate or N-methyl-D-aspartate (NMDA). We found that the NMDA-induced neurotoxicity was inhibited more effectively and potently than the glutamate-induced toxicity. Moreover, XR was found to exert mild inhibition of lipid peroxidation induced by $Fe^{2+}$/ascorbic acid in rat brain homogenates and some 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity. Taken together, these results demonstrate neuroprotective and antioxidant effects of XR, showing inhibition of oxidative and excitotoxic damage in the cultured cortical neurons, as well as inhibition of lipid peroxidation and its radical scavenging activity. Considering that excitotoxicity and oxidative stress pl ay crucial roles in neuronal cell damage during ischemia and reperfusion, these results may provide pharmacological basis for its clinical usage to treat ischemic stroke.

Antioxidant and Neuronal Cell Protective Effects of Columbia Arabica Coffee with Different Roasting Conditions

  • Jeong, Ji Hee;Jeong, Hee Rok;Jo, Yu Na;Kim, Hyun Ju;Lee, Uk;Heo, Ho Jin
    • Preventive Nutrition and Food Science
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    • v.18 no.1
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    • pp.30-37
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    • 2013
  • In vitro antioxidant activities and neuronal cell protective effects of ethanol extract from roasted coffee beans were investigated. Colombia arabica coffee (Coffea arabica) green beans were roasted to give medium ($230^{\circ}C$, 10 min), city ($230^{\circ}C$, 12 min) and french ($230^{\circ}C$, 15 min) coffee beans. Total phenolics in raw green beans, medium, city and french-roasted beans were $8.81{\pm}0.05$, $9.77{\pm}0.03$, $9.92{\pm}0.04$ and $7.76{\pm}0.01$ mg of GAE/g, respectively. The content of 5-O-caffeoylquinic acid, the predominant phenolic, was detected higher in medium-roasted beans than others. In addition, we found that extracts from medium-roasted beans particularly showed the highest in vitro antioxidant activity on ABTS radical scavenging activity and FRAP assays. To determine cell viability using the MTT assay, extracts from medium- roasted beans showed higher protection against $H_2O_2$-induced neurotoxicity than others. Lactate dehydrogenase (LDH) leakage was also inhibited by the extracts due to prevention of lipid peroxidation using the malondialdehyde (MDA) assay from mouse whole brain homogenates. These data suggest that the medium-roasting condition to making tasty coffee from Columbia arabica green beans may be more helpful to human health by providing the most physiological phenolics, including 5-O-caffeoylquinic acids.

A Study on the Relationship between the Uptake of $^{67}Ga-citrate$ and the Incorporation Rate of $^3H-thymidine$ and Metabolism of Protein in the Rat Livers Treated with $CCl_4$ (사염화탄소투여(四鹽化炭素投與)후 백서간세포(白鼠肝細胞)에서 $^{67}Ga$섭취율(攝取率)과 $^3H-thymidine$ 결합율(結合率) 및 단백대사(蛋白代射)와의 관계(關係)에 관(關)한 연구(硏究))

  • Hong, Seong-Woon
    • The Korean Journal of Nuclear Medicine
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    • v.19 no.1
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    • pp.83-93
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    • 1985
  • The ability of $^{67}Ga$, administered carrier free as the citrate complex, to localize in human and animal tumors to an extent sufficient to permit visualization of the lesion by scanning is well established. However, neither the mechanism of $^{67}Ga$ uptake by tumors or inflammatory cells nor its relationship to cell type or to the biochemical status of the cell is yet understood. Author investigated the uptake of $^{67}Ga-citrate$ using subcellular tissue fractionation of rat livers treated with $CCl_4$ associated with the $^3H-thymidine$ incorporation rate to detect subcellular localization of $^{67}Ga$ and it's relationship in DNA synthesis. Large amounts of $^{67}Ga$ associated with the soluble portion of tissue homogenate rather than with isolated cell organelles and not related nuclei residue in the regenerating period after hepatocellular injury caused by $CCl_4$. The elevated uptake of $^{67}Ga$ in the livers of $CCl_4$ treated rats was also inhibited when protein synthesis was stopped by cyclohexamide. Thus protein and the soluble portion of issue homogenates seems to play an important role in the elevated uptake of $^{67}Ga$ in liver injury induced by $CCl_4$ treated rats.

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Proteomic Changes in Chick Brain Proteome Post Treatment with Lathyrus Sativus Neurotoxin, β-N-Oxalyl-L-α,β-Diaminopropionic Acid (L-ODAP): A Better Insight to Transient Neurolathyrism

  • Anil Kumar, D;Natarajan, Sumathi;Omar, Nabil A M Bin;Singh, Preeti;Bhimani, Rohan;Singh, Surya Satyanarayana
    • Toxicological Research
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    • v.34 no.3
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    • pp.267-279
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    • 2018
  • Neurolathyrism is a neurodegenerative disorder characterized by spastic paraplegia resulting from the excessive consumption of Lathyrus sativus (Grass pea). ${\beta}$-N-Oxalyl-L-${\alpha},{\beta}$-diaminopropionic acid (L-ODAP) is the primary neurotoxic component in this pea. The present study attempted to evaluate the proteome-wide alterations in chick brain 2 hr and 4 hr post L-ODAP treatment. Proteomic analysis of chick brain homogenates revealed several proteins involved in cytoskeletal structure, signaling, cellular metabolism, free radical scavenging, oxidative stress and neurodegenerative disorders were initially up-regulated at 2 hr and later recovered to normal levels by 4 hr. Since L-ODAP mediated neurotoxicity is mainly by excitotoxicity and oxidative stress related dysfunctions, this study further evaluated the role of L-ODAP in apoptosis in vitro using human neuroblastoma cell line, IMR-32. The in vitro studies carried out at $200{\mu}M$ L-ODAP for 4 hr indicate minimal intracellular ROS generation and alteration of mitochondrial membrane potential though not leading to apoptotic cell death. L-ODAP at low concentrations can be explored as a stimulator of various reactive oxygen species (ROS) mediated cell signaling pathways not detrimental to cells. Insights from our study may provide a platform to explore the beneficial side of L-ODAP at lower concentrations. This study is of significance especially in view of the Government of India lifting the ban on cultivation of low toxin Lathyrus varieties and consumption of this lentil.

Gossypin Protects Primary Cultured Rat Cortical Cells from Oxidative Stress- and $\beta$-Amyloid-Induced Toxicity

  • Yoon, Injae;Lee, Kwang-Heun;Choi, Jungsook
    • Archives of Pharmacal Research
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    • v.27 no.4
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    • pp.454-459
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    • 2004
  • The present study investigated the effects of gossypin, 3,3',4',5,7,8-hexahydroxyflavone 8-glucoside, on the toxicity induced by oxidative stress or $\beta$-amyloid ($A_{\beta}$) in primary cultured rat cortical cells. The antioxidant properties of gossypin were also evaluated by cell-free assays. Gossypin was found to inhibit the oxidative neuronal damage induced by xanthinelxanthine oxidase or by a glutathione depleting agent, D,L-buthionine (S,R)-sulfoximine. In addition, gossypin significantly attenuated the neurotoxicity induced by $A_{{\beta}(25-35)}$. Furthermore, gossypin dramatically inhibited lipid peroxidation initiated by $Fe^{2+}$ and ascorbic acid in rat brain homogenates. It also exhibited potent radical scavenging activity generated from 1 ,1-diphenyl-2-picrylhydrazyl. These results indicate that gossypin exerts neuroprotective effects in the cultured cortical cells by inhibiting oxidative stress- and $A_{\beta}$-induced toxicity, and that the antioxidant properties of gossypin may contribute to its neuroprotective actions.

Antioxidant and Neuroprotective Effects of Hesperidin and its Aglycone Hesperetin

  • Cho, Jung-Sook
    • Archives of Pharmacal Research
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    • v.29 no.8
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    • pp.699-706
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    • 2006
  • The present study evaluated antioxidant and neuroprotective activities of hesperidin, a flavanone mainly isolated from citrus fruits, and its aglycone hesperetin using cell-free bioassay system and primary cultured rat cortical cells. Both hesperidin and hesperetin exhibited similar patterns of 1,1-diphenyl-2-picrylhydrazyl radical scavenging activities. While hesperidin was inactive, hesperetin was found to be a potent antioxidant, inhibiting lipid peroxidation initiated in rat brain homogenates by $Fe^{2+}$ and L-ascorbic acid. In consistence with these findings, hesperetin protected primary cultured cortical cells against the oxidative neuronal damage induced by $H_2O_2$ or xanthine and xanthine oxidase. In addition, it was shown to attenuate the excitotoxic neuronal damage induced by excess glutamate in the cortical cultures. When the excitotoxicity was induced by the glutamate receptor subtype-selective ligands, only the N-methyl-D-aspartic acid-induced toxicity was selectively and markedly inhibited by hesperetin. Furthermore, hesperetin protected cultured cells against the $A_{{\beta}(25-35)}-induced$ neuronal damage. Hesperidin, however, exerted minimal or no protective effects on the neuronal damage tested in this study. Taken together, these results demonstrate potent antioxidant and neuroprotective effects of hesperetin, implying its potential role in protecting neurons against various types of insults associated with many neurodegenerative diseases.

Inhibitory Effects of Xiaoshuan Zaizao Wan on Excitotoxic and Oxidative Neuronal Damage Induced in Primary Cultured Rat Cortical Cells (일차 배양한 흰쥐 대뇌피질세포의 흥분성 및 산화적 신경세포손상에 대한 소전재조환의 억제효과)

  • 조정숙
    • YAKHAK HOEJI
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    • v.47 no.6
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    • pp.369-375
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    • 2003
  • Xiaoshuan Zaizao Wan (XZW) has been used in China to improve hemiplegia, deviation of eye and mouth, and dysphasia due to cerebral thrombosis. To characterize pharmacological actions of XZW, we evaluated its effects on neuronal cell damage induced in primary cultured rat cortical cells by various oxidative insults, glutamate or N-methyl-D-aspartate (NMDA), and $\beta$-amyloid fragment ($A_{\beta(25-35)}$). XZW was found to inhibit the oxidative neuronal damage induced by $H_2O_2$, xanthine/xanthine oxidase, or $Fe^{2+}$/ascorbic acid. It also attenuated the excitotoxic damage induced by glutamate or NMDA. The NMDA-induced neurotoxicity was more effectively inhibited than the glutamate-induced toxicity. In addition, we found that XZW protected neurons against the $A_{\beta(25-35)}$-induced toxicity. Moreover; XZW exhibited dramatic inhibition of lipid peroxidation in rat brain homogenates and mild 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity. Taken together; these results demonstrate that XZW exerts neuroprotective effects against oxidative, excitotoxic, or $A_{\beta(25-35)}$-induced neuronal damage. These findings may provide pharmacological basis for its clinical usage treating the sequelae caused by cerebral thrombosis. Furthermore, XZW may exert beneficial effects on Alzheimer's disease and other oxidative stress-related neurodegenerative disorders.

Detection of viral pathogens and isolation of porcine circovirus 2 from postweaning multisystemic wasting syndrome-affected piglets (이유자돈 전신소모성증후군 이환 자돈에서의 바이러스성 원인체 검색 및 porcine circovirus 2 분리동정)

  • Park, Choi-Kyu;Kim, Hyun-Soo
    • Korean Journal of Veterinary Research
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    • v.44 no.4
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    • pp.561-569
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    • 2004
  • To detect viral agents and isolate porcine circovirus 2 (PCV2), 60 samples of lung and lymph node were collected from 5 to 12 week-old pigs that had showed clinical signs of postweaning multisystemic wasting syndrome (PMWS). Polymerase chain reactions (PCRs) were conducted to identify the viral pathogens including PCV1, PCV2, porcine parvovirus (PPV) and porcine reproductive and respiratory syndrome virus (PRRSV) that have been considered to be the causal agents of PMWS. Among 60 samples, PCV 2 was detected from 57 samples but no PCV 1 was detected. PRRSV and/or PPV were also detected from 27 (47.4%) samples and 1 (1.8%) sample of these 57 PCV 2-positive samples, respectively. Tissue homogenates were inoculated onto PCV-free PK-15 cell monolayers. Seven isolates were confirmed as PCV 2 by multiplex PCR, indirect immunofluorescence assay, and transmissible electron microscopy. These date suggest that PRRSV is a major cofactors causing PMWS in pigs that were infected with PCV2 in Korea.

Isolation and cultivation of swine encephalomyocarditis virus (돼지 뇌심근염 바이러스의 분리 배양)

  • Ha, Yong-kong;Yoon, Seok-min;Jung, Byung-tack;Park, Nam-yong;Lee, Bong-ju;Chung, Chi-young;Kee, Hye-young;Bae, Seong-yeol
    • Korean Journal of Veterinary Research
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    • v.31 no.4
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    • pp.479-484
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    • 1991
  • Encephalomyocarditis(EMC) virus was isolated from the mummified and stillborn pigs at a swine farm in Chonnam Province, experienced with EMC infection over the period Oct.~Dec. of 1989. In addition some cultural, serological properties of the isolates and experimental infections in the piglets were studied. The results obtained were as follows; 1. Two EMC virus strains with HA titers and CPE similar to EMC-ATCC were established in a baby hamster kidney (BHK)-21 cell line by inoculating homogenates of brain and heart of the 19 mummified or stillborn pigs and designated $K_3$ and $K_{11}$. 2. At the second BHK-21 cell line passage of the initial isolates CPE appeared after incubation for 16~18 hours, while at the fourth and fifth passage the highest titer of HA was recorded, titer of HA using rat and guinea pig erythrocytes. 3. One pig inoculated with the isolate $K_3$ showed dyspnea as clinical signs and died at the 10 days after inoculation at necropsy white necrotic foci were observed from the dead animal heart. 4. Although all the rest surviving pigs showed increases in antibody titer and body temperature of $40^{\circ}C$ above for the initial 2~4 days followed by the return to normal, there were no gross lesions when the animals were sacrificed at the 2 weeks after inoculation.

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Study on the pathogenesis of the piglets experimentally infected with Korean isolate of Aujeszky′s disease virus I. Histopathologic and electron microscopic observation (Aujeszky's disease virus 국내분리주 접종자돈의 병리발생에 관한 연구 I. 병리학적 및 전자현미경적 관찰)

  • 조우영;조성환;김재훈;박최규;황의경;조부제;정운선
    • Korean Journal of Veterinary Service
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    • v.19 no.1
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    • pp.1-29
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    • 1996
  • This study was conducted to elucidate the pathogenesis of Aujeszky's disease virus(ADV) by histopathologic examination. The first Korean ADV Isolate, which was isolated from piglets with clinical signs of Aujeszky's disease in Yangsan(YS) county, Kyungnam province, was inoculated into 32 days old piglets with a dose of $10^{5.9}$$TCID_{50}/ml$ through intranasal or intramuscular route. These piglets were sacrificed at intervals of every 24hrs for 8 days. The virulence of YS strain was determined by the observation of clinical signs, gross findings, and histopathologic changes in tissues. The virus recovery test was performed from brain, spleen, lung and tonsil in cell culture. The pathogenesis of YS strain was determined by the observation of histopathologlc lesions in CNS and neuronal tracts. The major clinical signs were fever, anorexia, dyspnea, constipation, tremor, ataxia, circling movement, hindleg paralysis and salivation. The clinical signs were more severe in piglets of the group inoculated intranasally than those of the intramuscularly inoculated gorup. Lymphocytopenia was detected on day 5 to day 6 postinoculation (PI). The ADV was recovered from the tissue homogenates of tonsil, lung, spleen and cerebrum in cell culture. The highest virus titer was detected from tonsil between day 6 and day 7 PI. Reddish sublobar consolidation foci were scattered in the apical and cardiac lobes of lung. Although yellowish necrotic foci were detected in tonsil and liver, hemorrhagic lesions were mainly observed in heart, kidney and lymph nodes. Histopathologically, degeneration and necrosis of nerve cells, nonsuppurative meningoe-ncephalitis, nodular gliosis and perivascular cuffings were observed in CNS. Multifocal fibronecrotic foci were observed in lung, liver, lymph nodes and spleen. The major pathologic changes were detected in the midbrain, pons and medulla oblongata. Eosinophilic intranuclear inclusion bodies were mainly observed in epithelia and /or macrophages of tonsil, liver, lung, spleen and submandibular lymph nodes, and neurons of brain, respectively. Observation of viral particles at various stages of replication were possible from the endothelial cells of the alveolar capillaries and tonsillar crypt epithelia by transmission electron microscope.

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