• The Korean Journal of Zoology
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• v.36 no.4
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• pp.439-451
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• 1993

Protein kinase C isozymes in a human prostate adenocarcinoma PC-3 cell line were characterized. Immunoreactive bands and immunocytochemical stains were obsenred in PC-3 cells with antibodies raised against protein kinase C ${\alpha}$, ${\beta}$, ${\gamma}$, $\delta$, $\varepsilon$, and ζ types, respectively. Protein kinase C ${\alpha}$ corresponded to a immunoreactive band at a molecular weight of 80,000-dalton, whereas molecular weights of other immunoreactive isozvmes of protein kinase C were detected at 68,000-dalton. Protein kinHse C $\delta$ and ζ antibodies detected additional bands at 55,000-dalton and 80,000-dalton, respectively Immunocvtochemical study confirmed the results of the immunoblotting experiments qualitatively: all six protein kinase C isozymes were detected in the cytoplasm of PC-3 cells. Translocation of protein kinase C in PC-3 cells were also examined with phorbol 12-myristate 13-acetate (PMA), bryostatin 2, diolein, and 1-oleoyl-2-acetyl glycerol (OAG). Differential reactions of protein kinase C isozvmes to these activators were obsenred. When PC-3 cells were treated with 10mM bryostatin 2, protein kinase C isozyme u was translocated into the nucleus, whereas s type was translocated into the plasma membrane and the nucleus. Protein kinase C ${\alpha}$ and ζ types were translocated into the nucleus following the treatment with 101M diolein, whereas protein kinase C ${\alpha}$, ${\beta}$, ${\gamma}$, and $\varepsilon$ types were translocated into the nucleus by the treatment with 10mM OAG. Protein kinase C ${\alpha}$ and $\varepsilon$ types were translocated into the nucleus in the presence of 100nM PMA. Protein kinase C $\delta$ type was translocated to the nuclear membrane by these activators, however, only PMA-induced translocation was inhibited by protein kinase C inhibitor, 1-(5-isoquinolinesulfonyll-2-methvlpiperazine dihvdrochloride (H7) . H7 inhibited translocation of protein kinase C ${\alpha}$ type induced by PMA, ${\beta}$ type by OAG and s type by PMA and OAG, whereas it did not affect translocations induced by bryostatin and diolein, respectively. These results suggest that there exist six isoformes of protein kinase C (${\alpha}$, ${\beta}$, ${\gamma}$, $\delta$, $\varepsilon$ and ζ types) in PC-3 cells and that each of these isozvmes distinctivelv reacts to bryostatin, diolein, OAG and PMA, in part due to an altered molecular size and conceivably discrete binding site(s).

• The Journal of Korean Medicine
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• v.27 no.2 s.66
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• pp.122-133
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• 2006

Objectives : The amyloid $\beta$-protein ($A\beta$) is the principal component of the senile plaques characteristic of Alzheimer's disease (AD) and elicits a toxic effect on neurons in vitro and in vivo. Many environmental factors including antioxidants and proteoglycans modify $A{\beta}toxicity$. In this study, we have investigated the protective effects of water- and organic solvent-extracts of Hemerocallis fulva root fractions pre-extracted with methanol on $A\beta$-induced oxidative cell death in cultured rat pheochromocytoma (PC12) cells. Methods : For this study, we used MTT reduction assay for detection of protective effects of water- and organic solvent-extracts of Hemerocallis fulva root fractions pre-extracted with methanol on $A{\beta}_{25-35}$-induced cytotoxicity to PC12 cells. We also used cell-based $\beta$-secretase assay system to investigate the inhibitory effect of water- and organic solvent-extracts of Hemerocallis fulva root on $\beta$-secretase activity. Results : We previously reported that methanol extracts of Hemerocallis fulva root strongly attenuated cytotoxicity induced by the three $A\beta$ fragments ($A{\beta}_{25-35},\;A{\beta}_{1-42}\;A{\beta}_{1-43}$) to both SK-N-MC and PC12 cells. In the present study, we found that butanol-, ethylacetate-, chloroform-, and water-extracts of Hemerocallis fulva root fractions pre-extracted with methanol had strong protective effects against $A{\beta}_{25-35}$-induced cytotoxicity to PC12 cells and inhibitory potency to $\beta$-secretase activity. Conclusion : These results suggest that butanol-, ethylacetate-, chloroform-, and water-extracts of Hemerocallis fulva root fractions pre-extracted with methanol may contain the protective component(s) against $A\beta$-induced cell death in PC12 cells as well as inhibitory component(s) to $\beta$-secretase activity.

• Journal of Oriental Neuropsychiatry
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• v.20 no.1
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• pp.1-19
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• 2009

Objectives : This Study was performed to assess the antioxidative and neuroprotective effect of Guibi-tang(Guipi-tang) and Guibi-tang gamibang(Guipitang jiaweifang) on PC12 cells. Methods : The antioxidative effect was investigated through the DPPH radical and ABTS cation scavenging methods and total polyphenol amount of Guibi-tang(Guipitang) and Guibi-tang gamibang(Guipitang jiaweifang). The neuroprotective effect of Guibi-tang(Guipitang) and Guibi-tang gamibang(Guipitang jiaweifang) was assessed using MTT assay in PC12 cells. The scavenging effect of Guibi-tang(Guipitang) and Guibi-tang gamibang(Guipitang jiaweifang) on NO and ROS production induced by 6-OHDA in PC12 cells was evaluated, as well as the attenuating effect of Guibi-tang gamibang(Guipitang jiaweifang) on GSH reduction. Results : 1. Guibi-tang(Guipitang) and Guibi-tang gamibang(Guipitang jiaweifang) had concentration-dependent scavenging activities of DPPH radical 2. Guibi-tang(Guipitang) and Guibi-tang gamibang(Guipitang jiaweifang) had concentration-dependent scavenging activities of ABTS cation. 3. Total polyphenol amount of Guibi-tang(Guipitang) and Guibi-tang gamibang(Guipitang jiaweifang) was calculated 79.10${\pm}$2.20 pg/IO mg and 121.03${\pm}$1.11 pg/IO mg, respectively. 4. Cell viability of Guibi-tang(Guipitang) was increased in a dose dependent manner. Guibi-tang gamibang(Guipitang jiaweifang) was increased at low concentrations, but decreased at high concentrations. 5. In Guibi-tang(Guipitang), cell viability of PC12 cell treated with 6-OHDA was decreased by pre-treatment, and increased by post- and co- treatment. Cell viability of Guibi-tang gamibang(Guipitang jiaweifang) showed variable effects by pre-treatment, but increased by post- and co- treatment. 6. NO production rate of Guibi-tang(Guipitang) didn't show a significant effect, but that of Guibi-tang gamibang(Guipitang jiaweifang) was decreased in a dose dependent manner. 7. ROS production rate of Guibi-tang(Guipitang) was decreased at some concentrations. In Guibi-tang gamibang(Guipitang jiaweifang), ROS production rate was decreased at high concentrations. 8. Guibi-tang gamibang(Guipitang jiaweifang) protected the 6-OHDA-induced GSH reduction. Conclusions : These results demonstrate that both Guibi-tang(Guipitang) and GBTGMB have antioxidative and neuroprotective effect, but Guibi-tang gamibang(Guipitang jiaweifang) has more antioxidative and neuroprotective effect than Guibi-tang.

• The Korean Journal of Food And Nutrition
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• v.33 no.2
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• pp.215-221
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• 2020

The purpose of this study was to investigate the protective effect on oxidative stress induced PC12 cells, and volatile flavor composition of essential oils derived from medicinal plant seeds- Gossypium hirsutum L. (G. hirsutum), Coix lachryma-jobi (C. lachryma-jobi) and Oenothera biennis (O. biennis). The essential oils were obtained by the solvent (hexane) extraction method from the seeds. The essential oils of the seeds were analyzed by the solid-phase micro-extraction gas chromatography mass spectrometry (SPME-GC/MS). The major compounds of G. hirsutum, C. lachryma-jobi and O. biennis were cyclonexanol (16.65%), β-asarone (14.29%) and ylangene (50.01%). The DPPH radical scavenging activity (IC₅₀) was the highest value of 8.52 mg/mL in the O. biennis. Additionally, IC₅₀ values of G. hirsutum and C. lachryma-jobi were 26.76 mg/mL and 36.81 mg/mL. For the oxidative stress on PC12 cells, we treated with hydrogen peroxide (H₂O₂). The pretreatment of oxidative stress induced PC12 cells with all the essential oils preserved or increased their cell viability and G. hirsutum and O. biennis attenuated the ROS generation (by 68.75% and 56.25% vs. H₂O₂ control). The results of this study suggest that the essential oils derived from medicinal plant seeds could be used as valuable back data as a natural essential oil material to prevent neurodegenerative diseases by protecting neuro-cells.

• Food Science and Preservation
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• v.18 no.3
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• pp.358-365
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• 2011

The antioxidant and neuronal cell-protective effects of hot water extract from commercial buckwheat tea (CBTE) were evaluated. The 2,2'-azino-bis(3-ethyl-benzthiazoline-6-sulfonic acid) (ABTS) radical scavenging activity, ferric reducing antioxidant power (FRAP), and malondialdehyde (MDA) inhibitory effect of the CBTE increased in a dose-dependent manner. The Intracellular reactive oxygen species (ROS) accumulation that resulted from hydrogen peroxide ($H_2O_2$) treatment more significantly decreased when CBTE was present in the media than when the PC12 cells were treated only with $H_2O_2$. In the neuronal cell viability assay using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazoliumbromide (MTT), the aqueous extracts showed a protective effect against $H_2O_2$-induced neurotoxicity, and the lactate dehydrogenase (LDH) release into the medium was also inhibited by CBTE. The total phenolics of CBTE was 9,608.10 mg/100 g, and the major phenolic compounds were rutin (13.42 mg/100 g) and quercitrin (0.90 mg/100 g). These data suggested that CBTE, including the aforementioned phenolics, may be useful in reducing the risk of neurodegenerative disease.

• Korean Journal of Food Science and Technology
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• v.46 no.3
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• pp.369-374
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• 2014

The physiological characteristics of kiwi (Actinidia delicosa) fruit were analyzed, which inclued its nutritional composition, in vitro-antioxidant activities, and neuronal cell protective effects. The most abundant components of mineral, amino acid, and fatty acid were found to be potassium (K), glutamic acid, and a-linolenic acid, respectively. The major free sugars were fructose, glucose, and sucrose. In addition, ${\beta}$-carotene and vitamin C contents were $1.35{\mu}g/100mL$ and 29.21 mg/100 g, respectively. The 2,2'-azinobis-(3-ethylbenothiazline-6-sulfonic acid) (ABTS) radical-scavenging activity of the n-hexane fraction obtained from the kiwi extract was 10.52% at a concentration of $1000{\mu}g/mL$. The malondialdehyde (MDA) inhibition of the n-hexane fraction was found to increased in a dose-dependent manner. The intracellular reactive oxygen species (ROS) accumulation after hydrogen peroxide ($H_2O_2$) treatment of PC12 cells was significantly reduced in the presence of the n-hexane fraction compared to PC12 cells treated with $H_2O_2$ only. Moreover, in the a MTT assay, the n-hexane fraction showed in vitro-protective effects against $H_2O_2$-induced neurotoxicity.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.7
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• pp.938-947
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• 2016

Antioxidant activities and neuroprotective effects of ethyl acetate fraction from Dendropanax morbifera (EFDM) against high glucose-induced oxidative stress and neurotoxicity were investigated to confirm their physiological activities. An 80% ethanolic extract of D. morbifera showed the highest contents of total phenolic compounds as well as 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) and 1,1-diphenyl-2-picrylhydrazyl radical scavenging activities. The extract was fractionated using several solvents, and the ethyl acetate fraction showed the highest activities in ferric reducing/antioxidant power and malondialdehyde inhibitory assays. To evaluate the neuroprotective effect based on antioxidant activities, cell viability was assessed using PC12 and MC-IXC cells in $H_2O_2$- and high glucose-induced cytotoxic assays, respectively. EFDM evidently showed neuroprotective effects in all cells (neuron-like PC12 cells and human brain-originated neuroblastoma MC-IXC cells). Inhibitory effect of the extract on acetylcholinesterase (AChE) as an acetylcholine-hydrolyzing enzyme was performed to examine the effect on cognitive function. EFDM presented an AChE inhibitory effect. Finally, high-performance liquid chromatography analysis showed that the major phenolic compound of EFDM is probably a rutin.

• Korean Journal of Food Science and Technology
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• v.34 no.3
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• pp.505-509
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• 2002

In the course of screening program for VHR DS-PTPase (dual-specificity protein tyrosine phosphatase) from natural sources, Gastrodia elata was selected. One compound showing potent inhibitory activity was isolated by the solvent extraction and column chromatography including silica gel, ODS RP-18, Sephades LH-20, and HPLC. This compound was identified as baicalein by several NMR techniques such as $^1H-NMR$, $^{13}C-NMR$, and DEPT. Baicalein showed selective inhibitory activity against VHR DS-PTPase with $IC_{50}=2.4\;{\mu}M$, and showed cytotoxicity against several human cancer cell lines with an $GI_{50}$ of $5.26{\sim}12.93\;{\mu}g/mL$ range, including, melanoma (LOX-IMVI), lung cancer (NCI H23 and A549), colon cancer (HCT 116 and SW 620), prostate cancer (PC-3), and leukemia (MOLT 4F).

• Journal of agriculture & life science
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• v.44 no.2
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• pp.57-66
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• 2010

The antioxidant and neuronal cell protective effects of water extract from purple sweet potato were investigated. The total phenolics and monomeric anthocyanin contents of purple sweet potato extract were 44.25 mg/g and 2,394 mg/L, respectively. The antioxidant activities of purple sweet potato extract were evaluated using various antioxidant tests, including 1,1-diphenyl- 2-picrylhydrazyl (DPPH), 2,2'-azino- bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical scavenging activities, ferric reducing/antioxidant power (FRAP) and reducing power. In these assays, the extract of purple sweet potato presented significant radical scavenging activities, FRAP, and reducing power in a dose-dependent manner. MTT {3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyl- tetrazoliumbromide} reduction assay showed significantly increase in cell viability when PC12 cells were pretreated with purple sweet potato extract. Because oxidative stress is also known to increase neuronal cell membrane breakdown, we further investigated by lactate dehydrogenase (LDH) and neutral red uptake assay. Purple sweet potato extract inhibited oxidative stress-induced membrane damage in neuronal cells. Therefore, these data results demonstrated that water extract of purple sweet potato have antioxidant activity and neuronal cell protective effect thus it has great potential as a natural source for human health.

• Tuberculosis and Respiratory Diseases
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• v.40 no.3
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• pp.213-222
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• 1993

Background : Bronchial asthma has been known as an inflmmatory disease. There have been many evidences that polymorphonuclear leukocytes (PMN) might play an important role in the pathogrnesis of asthma. Although many investigators suggested that pneumocyte injury by PMN-derived oxygen radicals may contribute to the pathogenesis of asthma, there has been few report for a direct evidence of oxygen radicals-mediated pneumocyte injury in bronchial asthma. Furthermore the exact mechanism of oxygen radicals-mediated pneumocyte injury is still controversy. This study was designed to establish a direct in vitro evidence and its clinical significance of pneumocyte injury by PMN-derived superoxide anion in bronchial asthma and to elucidate the main mechanism of superoxide anion-mediated pneumocyte injury. Methods : 12 stable asthmatics and 5 healthy volunteers were participated in this study. PMN was separated from peripheral venous blood samples by using dextran sedimentation and Ficoll-Hypaque density gradient separation method. Superoxide anion productions by PMN and plasma SOD activities were measured by spectrophotometric assay using the principle of SOD inhibitable cytochrome c reduction. PMN-mediated pneumocyte injuries were measured by $^{51}Cr$-release assay using A549 pneumocytes and were expressed as percent lysis and percent detachment. Results: 1) PMN from asthmatics produced more amount of superoxide anion compared to PMN from normal subjects ($6.65{\pm}0.58$ vs $2.81{\pm}0.95\;nmol/1{\times}10^6$ cells, p<0.05), and showed an inverse correlation with $FEV_1$(R=-0.63, p<0.05), but no correlation with $PC_{20}$ histamine in asthmatics. 2) Plasma SOD activities were decreased in asthmatics compared to normal subjects but not significant, and showed a positive correlation with $FEV_1$(R=0.63, p<0.05) but no correlation with $PC_{20}$ histamine in asthmatics. 3) There were a positive correlation between plasma SOD activity and superoxide anion production by PMN in normal subjects (R=0.88, p<0.05) but not in asthmatics. 4) PMN-mediated pneumocyte injury was predominantly expressed as cell detachment rather than cell lysis in both groups, and PMN from asthmatics showed more potent cytotoxic effect on A549 pneumocytes compated to PMN from normal subjects. PMN-mediated detachment rather than lysis of A549 pneumocytes was significantly inhibited by in vitro SOD but not by diluted serum. 5) PMN-mediated detachment rather than lysis of A549 pneumocytes showed a good correlation with superoxide anion production by PMN (R=0.90 in normal subjects, R=0.82 in asthmatics, p<0.05) but no correlation with plasma SOD activity. PMN-mediated pneumocyte injuries were not correlated with $FEV_1$ or $PC_{20}$ histamine in asthmatics. 6) There were no significant differences in PMN-mediated pneumocyte injuries between allergic and nonallergic asthmatics. Conclusion : Our results suggest that pneumocyte injury by PMN-derived superoxide anion may partially contribute to the pathogenesis of asthma and that cell detachment rather than cell lysis may be the mechanism of superoxide anion-mediated pneumocyte injury.