• Title/Summary/Keyword: PC12 cells

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Sea Buckthorn (Hippophae rhamnoides L.) Leaf Extracts Protect Neuronal PC-12 Cells from Oxidative Stress

  • Cho, Chi Heung;Jang, Holim;Lee, Migi;Kang, Hee;Heo, Ho Jin;Kim, Dae-Ok
    • Journal of Microbiology and Biotechnology
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    • v.27 no.7
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    • pp.1257-1265
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    • 2017
  • The present study was carried out to investigate the antioxidative and neuroprotective effects of sea buckthorn (Hippophae rhamnoides L.) leaves (SBL) harvested at different times. Reversed-phase high-performance liquid chromatography analysis revealed five major phenolic compounds: ellagic acid, gallic acid, isorhamnetin, kaempferol, and quercetin. SBL harvested in August had the highest total phenolic and flavonoid contents and antioxidant capacity. Treatment of neuronal PC-12 cells with the ethyl acetate fraction of SBL harvested in August increased their viability and membrane integrity and reduced intracellular oxidative stress in a dose-dependent manner. The relative populations of both early and late apoptotic PC-12 cells were decreased by treatment with the SBL ethyl acetate fraction, based on flow cytometry analysis using annexin V-FITC/PI staining. These findings suggest that SBL can serve as a good source of antioxidants and medicinal agents that attenuate oxidative stress.

Protective Effects of Black Soybean Seed Coat Extracts against Oxidative Stress-induced Neurotoxicity (산화적 손상에 의해 유도된 신경세포독성에 대한 검정콩 껍질 추출물의 보호효과)

  • Kwak, Ji Hyeon;Jo, Yu Na;Jeong, Ji Hee;Kim, Hyeon Ju;Jin, Su Il;Choi, Sung-Gil;Heo, Ho Jin
    • Korean Journal of Food Science and Technology
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    • v.45 no.2
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    • pp.257-261
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    • 2013
  • Rat pheochromocytoma cells (PC12) and mice were utilized as in vitro and in vivo models to determine the neuroprotective effects of a 70% acetone extract of black soybean seed coat (BSSCE). BSSCE showed higher total phenolic contents than other extracts. Intracellular reactive oxygen species accumulation from $H_2O_2$ treatment of PC12 cells was significantly reduced when BSSCE was present in the media compared to PC12 cells treated with $H_2O_2$ only. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium-bromide (MTT) reduction assay and lactate dehydrogenase assay also showed significantly increased protective effects in PC12 cells. In addition, BSSCE improved the in vivo cognitive ability against amyloid beta peptide-induced neuronal deficits.

The Antioxidative and Neuroprotective Effect of Guibi-tang(Guipitang) and Guibi-tang gamibang(Guipitang jiaweijang) on PC12 cells. (귀비탕(歸脾湯)과 귀비탕가미방(歸脾湯加味方)의 항산화 효과 및 6-Hydroxydopamine에 대한 PC12 세포 보호효과 비교연구)

  • Lim, Jae-Whan;Kim, Jong-Woo;Chung, Sun-Yong;Cho, Sung-Hoon;Oh, Myung-Sook;Hwang, Wei-Wan
    • 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.

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Anti-tumor activities of Panax quinquefolius saponins and potential biomarkers in prostate cancer

  • He, Shan;Lyu, Fangqiao;Lou, Lixia;Liu, Lu;Li, Songlin;Jakowitsch, Johannes;Ma, Yan
    • Journal of Ginseng Research
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    • v.45 no.2
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    • pp.273-286
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    • 2021
  • Background: Prostate carcinoma is the second most common cancer among men worldwide. Developing new therapeutic approaches and diagnostic biomarkers for prostate cancer (PC) is a significant need. The Chinese herbal medicine Panax quinquefolius saponins (PQS) have been reported to show anti-tumor effects. We hypothesized that PQS exhibits anti-cancer activity in human PC cells and we aimed to search for novel biomarkers allowing early diagnosis of PC. Methods: We used the human PC cell line DU145 and the prostate epithelial cell line PNT2 to perform cell viability assays, flow cytometric analysis of the cell cycle, and FACS-based apoptosis assays. Microarray-based gene expression analysis was used to display specific gene expression patterns and to search for novel biomarkers. Western blot and quantitative real-time PCR were performed to demonstrate the expression levels of multiple cancer-related genes. Results: Our data showed that PQS inhibited the viability of DU145 cells and induced cell cycle arrest at the G1 phase. A significant decrease in DU145 cell invasion and migration were observed after 24 h treatment by PQS. PQS up-regulated the expression levels of p21, p53, TMEM79, ACOXL, ETV5, and SPINT1 while it down-regulated the expression levels of bcl2, STAT3, FANCD2, DRD2, and TMPRSS2. Conclusion: PQS promoted cells apoptosis and inhibited the proliferation of DU145 cells, which suggests that PQS may be effective for treating PC. TMEM79 and ACOXL were expressed significantly higher in PNT2 than in DU145 cells and could be novel biomarker candidates for PC diagnosis.

Protective Effects of Hwansodan(Huanshao-dan) Water Extract in Serum Deprivation-induced Apoptosis of PC12 Cells (환소단이 영양혈청 결핍성 PC12 신경세포의 apoptosis에 미치는 영향)

  • 임준식;김명선;소홍섭;이지현;한상혁;허윤;박래길;문병순
    • The Journal of Korean Medicine
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    • v.21 no.4
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    • pp.64-72
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    • 2000
  • Objectives : This study was designed to investigate the neuroprotective effect of Hwansodan(Huanshao-dan) on the apoptosis induced by withdrawal of neurotrophic support. Methods : PCl2 pheochromocytoma cells have been used extensively as a model for studying the cellular and molecular effects of neuronal cells. The viability of cells was measured by MTT assay. We used DNA fragmentation and caspase 3-like protease activation assay. Results : The water extract of Hwansodan(Huanshao-dan) significantly showed protective effects on serum and glucose deprivation-induced apoptotic death. Hwansodan(Huanshao-dan) also prevents DNA fragmentation and caspase 3-like protease activation, representing typical neuronal apoptotic phenomena in PCl2 pheochromocytoma cells and induces tyrosine phosphorylation of proteins around 44 kDa, which was identified as ERK1 with electrophoretic gel mobility shift by Western blot. In addition, MAPK kinase(MEK) inhibitor PD98059 and Ras inactivator, ${\alpha}-hydroxyfarnesylphosphonic$ acid attenuated the neuroprotective effects of Hwansodan(Huanshao-dan) in serum-deprived PCl2 cells. Conclusions : These results indicate that Ras/MEK/ERK signaling pathway plays a key role in neuroprotective effects of Hwansodan(Huanshao-dan) in serum-deprived PCl2 cells. Taken together, we suggest the possibility that Hwansodan(Huanshao-dan) might provide a neurotrophic-like activity in PCl2 cells.

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Neuritogenic activity of hot water extract from Hericium erinaceus

  • Li, Hua;See, Hye-Jung;Moon, BoKyung;Yoo, Young-Bok;Lee, Chan
    • Journal of Mushroom
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    • v.11 no.3
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    • pp.117-123
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    • 2013
  • Hot water soluble extract was prepared from Hericium erinaceus and its neuritogenic activity on PC12h cells was analyzed, which is a clone originating from a rat pheochromocytomon. The moisture content of freeze dried hot water extract was 12.08%. The extract was mainly composed of carbohydrate (51.24%) followed by crude protein (24.04%), crude fat (0.26%), dietary fiber (5.09), and ash (12.18%). Fatty acids, glucan and inorganic constituents were found as minor components. The neuritogenic activity of hot water extract was evaluated under microscopic observation of neurite outgrowth in PC12h cells and by measuring the neurite length of induvidual cell. The extract exhibited strong effect of neurite outgrowth in a dose-dependent manner from 0.01 mg/mL to 1 mg/mL, in which longer neurite outgrowth was observed as the treatment dose increased.

Nuclear Rac1 regulates the bFGF-induced neurite outgrowth in PC12 cells

  • Kim, Eung-Gook;Shin, Eun-Young
    • BMB Reports
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    • v.46 no.12
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    • pp.617-622
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    • 2013
  • Rac1 plays a key role in neurite outgrowth via reorganization of the actin cytoskeleton. The molecular mechanisms underlying Rac1-mediated actin dynamics in the cytosol and plasma membrane have been intensively studied, but the nuclear function of Rac1 in neurite outgrowth has not yet been addressed. Using subcellular fractionation and immunocytochemistry, we sought to explore the role of nuclear Rac1 in neurite outgrowth. bFGF, a strong agonist for neurite outgrowth in PC12 cells, stimulated the nuclear accumulation of an active form of Rac1. Rac1-PBR (Q) mutant, in which six basic residues in the polybasic region at the C-terminus were replaced by glutamine, didn't accumulate in the nucleus. In comparison with control cells, cells expressing this mutant form of Rac1 displayed a marked defect in extending neurites that was concomitant with reduced expression of MAP2 and MEK-1. These results suggest that Rac1 translocation to the nucleus functionally correlates with bFGF-induced neurite outgrowth.

Octyl Gallate Inhibits ATP-induced Intracellular Calcium Increase in PC12 Cells by Inhibiting Multiple Pathways

  • Guo, Yujie;Hong, Yi-Jae;Jang, Hyun-Jong;Kim, Myung-Jun;Rhie, Duck-Joo;Jo, Yang-Hyeok;Hahn, Sang-June;Yoon, Shin-Hee
    • The Korean Journal of Physiology and Pharmacology
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    • v.14 no.1
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    • pp.21-28
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    • 2010
  • Phenolic compounds affect intracellular free $Ca^{2+}$ concentration ($[Ca^{2+}]_i$) signaling. The study examined whether the simple phenolic compound octyl gallate affects ATP-induced $Ca^{2+}$ signaling in PC12 cells using fura-2-based digital $Ca^{2+}$ imaging and whole-cell patch clamping. Treatment with ATP ($100\;{\mu}M$) for 90 s induced increases in $[Ca^{2+}]_i$ in PC12 cells. Pretreatment with octyl gallate (100 nM to $20\;{\mu}M$) for 10 min inhibited the ATP-induced $[Ca^{2+}]_i$ response in a concentration-dependent manner ($IC_{50}=2.84\;{\mu}M$). Treatment with octyl gallate ($3\;{\mu}M$) for 10 min significantly inhibited the ATP-induced response following the removal of extracellular $Ca^{2+}$ with nominally $Ca^{2+}$-free HEPES HBSS or depletion of intracellular $Ca^{2+}$ stores with thapsigargin ($1\;{\mu}M$). Treatment for 10 min with the L-type $Ca^{2+}$ channel antagonist nimodipine ($1\;{\mu}M$) significantly inhibited the ATP-induced $[Ca^{2+}]_i$ increase, and treatment with octyl gallate further inhibited the ATP-induced response. Treatment with octyl gallate significantly inhibited the $[Ca^{2+}]_i$ increase induced by 50 mM KCI. Pretreatment with protein kinase C inhibitors staurosporin (100 nM) and GF109203X (300 nM), or the tyrosine kinase inhibitor genistein ($50\;{\mu}M$) did not significantly affect the inhibitory effects of octyl gallate on the ATP-induced response. Treatment with octyl gallate markedly inhibited the ATP-induced currents. Therefore, we conclude that octyl gallate inhibits ATP-induced $[Ca^{2+}]_i$ increase in PC12 cells by inhibiting both non-selective P2X receptor-mediated influx of $Ca^{2+}$ from extracellular space and P2Y receptor-induced release of $Ca^{2+}$ from intracellular stores in protein kinase-independent manner. In addition, octyl gallate inhibits the ATP-induced $Ca^{2+}$ responses by inhibiting the secondary activation of voltage-gated $Ca^{2+}$ channels.

The Effects of Hibiscus syriacus(HSS) Extract on the Alzheimer's Disease Model Induced by CT-105 and ${\beta}A$ (목근피(木槿皮)가 CT105와 ${\beta}A$로 유도된 Alzheimer's Disease 병태(病態) 모델에 미치는 영향)

  • Choi, Byung-Man;Jung, In-Chul;Lee, Sang-Ryong
    • Journal of Oriental Neuropsychiatry
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    • v.15 no.2
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    • pp.119-139
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    • 2004
  • This research investigates the effect of the Hibiscus syriacus(HSS) on Alzheimer's disease. Specifically, the effects of the HSS extract on (1) $IL-1{\beta}$, IL-6, and $TNF-{\alpha}$ mRNA of PC-12 cells treated with LPS; (2) amyloid precursor proteins(APP), acetylcholinesterase(AChE), and glial fibrillary acidic protein(GFAP) mRNA of PC-12 cells treated with CT-105; (3) the AChE activity and the APP production of PC-12 cell treated with CT-105; (4) the behavior; (4) expression of $IL-1{\beta}$, $TNF-{\alpha}$, $IL-1{\beta}$ mRNA, $TNF-{\alpha}$ mRNA, and reactive oxygen species(ROS); (5) the infarction area of the hippocampus, and brain tissue injury in Alzheimer's diseased mice induced with ${\beta}A$ were investigated. The results were summarized below ; 1. The HSS extract suppressed the expression of $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ mRNA in THP-l cells treated with LPS. 2. The HSS extract suppressed the expression of APP, AChE, and GFAP mRNA in PC-12 cells treated with CT-105. 3. The HSS extract suppressed the AChE activity, and the production of APP significantly in PC-12 cells treated with CT-105. 4. For the HSS extract group a significant inhibitory effect on the memory deficit was shown for the mice with Alzheimer's disease induced by ${\beta}A$ in the Morris water maze experiment, which measured stop-through latency, and distance movement-through latency. 5. The HSS extract suppressed the over-expression of $IL-1{\beta}$, $TNF-{\alpha}$, $IL-1{\beta}$ and $TNF-{\alpha}$ mRNA, CD68/GFAP, ROS in the mice with Alzheimer's disease induced by ${\beta}A$. 6. The HSS extract reduced the infarction area of hippocampus, and controlled the injury of brain tissue in the mice with Alzheimer's disease induced by ${\beta}A$. These results suggest that the HSS extract may be effective for the prevention and treatment of Alzheimer's disease. Investigation into the clinical use of the HSS extract for Alzheimer's disease is suggested for future research.

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The Effects of Chaenomelis fructus,(CMF) Extract on the Alzheimer's Disease Model Induced by CT-105 and ${\beta}A$ (목과(木瓜)가 CT105 와 ${\beta}A$로 유도(誘導)된 Alzheimer's Disease병태(病態) 모델에 미치는 영향(影響))

  • Kim, Myung-Jin;Jung, In-Chul;Lee, Sang-Ryong
    • Journal of Oriental Neuropsychiatry
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    • v.16 no.1
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    • pp.97-117
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    • 2005
  • This research investigates the effect of the Chaenomelis fructus(CMF) on Alzheimer's disease. Specifically, the effects of the CMF extract on (1) >$IL-1{\beta}$, IL-6, and $TNF-{\alpha}$ mRNA of PC-12 cells treated with LPS; (2) amyloid precursor proteins(APP), acetylcholinesterase(AChE), and glial fibrillary acidic protein(GFAP) mRNA of PC-12 cells treated with CT-105; (3) the AChE activity and the APP production of PC-12 cell treated with CT-105; (4) the behavior of AD mice with ${\beta}A$; (5) expression of $IL-1{\beta}$, $TNF-{\alpha}$, MDA, $IL-1{\beta}$ mRNA, $TNF-{\alpha}$ mRNA, and ROS; (6) the infarction area of the hippocampus, and brain tissue injury in Alzheimer's diseased mice induced with ${\beta}A$ were investigated. The results were summarized as follows; 1. The CMF extract suppressed the expression of $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ mRNA in THP-1 cells treated with LPS. 2. The CMF extract suppressed the expression of APP, AChE, and GFAP mRNA in PC-12 cells treated with CT-105. 3. The CMF extract suppressed the AChE activity, and the production of APP significantly in PC-12 cells treated with CT-105. 4. A significant inhibitory effect on the memory deficit was shown on the CMF extract group of the mice with Alzheimer's disease induced by ${\beta}A$ in the Morris water maze experiment, which measured stop-through latency, and distance movement-through latency. 5. The CMF extract suppressed the over-expression of $IL-1{\beta}$ protein, $TNF-{\alpha}$ protein, MDA, $IL-1{\beta}$ protein, mRNA, $TNF-{\alpha}$ mRNA, CD68/GFAP, and ROS in the mice with Alzheimer's disease induced by ${\beta}A$. 6. The CMF extract reduced the infarction area of hippocampus, and controlled the injury of brain tissue in the mice with Alzheimer’s disease induced by ${\beta}A$. These results suggest that the CMF extract may be effective for the treatment of Alzheimer’s disease. Investigation into the clinical use of the CMF extract for Alzheimer's disease is suggested for future research.

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