• Title/Summary/Keyword: PC12 cells

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Effects of an Aqueous Extract of Asparagus cochinchinensis on the Regulation of Nerve Growth Factor in Neuronal Cells (신경세포에서 신경성장인자(nerve growth factor)의 조절에 미치는 천문동(Asparagus cochinchinensis) 열수추출물의 영향)

  • Lee, Hyun Ah;Kim, Ji Eun;Song, Sung Hwa;Sung, Ji Eun;Jung, Min Gi;Kim, Dong Seob;Son, Hong Joo;Lee, Chung Yeoul;Lee, Hee Seob;Hwang, Dae Youn
    • Journal of Life Science
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    • v.26 no.5
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    • pp.509-518
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    • 2016
  • Asparagus cochinchinensis is a medical plant that has long been used to treat fever, cough, kidney disease, breast cancer, inflammatory disease and brain disease in northeast Asian countries. Although several studies have been conducted on the anti-neuroinflammatory effects of A. cochinchinensis, the correlation between these effects and nerve growth factor (NGF) has not yet been examined. In this study, we investigated the effects of an aqueous extract of A. cochinchinensis (AEAC) on the secretion and action mechanism of NGF in neuronal cells. The concentration of the NGF protein in the supernatant collected from cultured cells increased significantly in B35 cells treated with AEAC in comparison with the vehicle-treated group without any specific cytotoxicity. Furthermore, the mRNA expression of NGF showed a very similar pattern to its protein concentration. To examine the bioactivity of NGF secreted from B35 cells, undifferentiated PC12 cells were cultured in an AEAC-conditioned medium and neuritic outgrowth was observed. The dendrite length of PC12 cells in the AEAC-treated group was significantly higher than that in the vehicle-treated group. Moreover, the level of the downstream effectors p-TrkA and p-ERK of the high-affinity NGF receptor was significantly higher in the AEAC-treated group, while the expression of the downstream effectors of the low-affinity NGF receptor was significantly lower in the same group. These results suggest that AEAC may contribute to the regulation of NGF expression and secretion in neuronal cells; it is therefore an excellent candidate for further investigation as a therapeutic drug for neurodegenerative diseases.

The Changes of Growth Patterns and the Production of Brain-Derived Neurotrophic Factors (BDNFs) in Perfusion Cultivation of Human Neuroblastoma Cells

  • Hong, Jong-Soo;Lee, Joo-Nho;Kim, Sun-Hee;Park, Kyung-Yoo;Cho, Jin-Sang;Lee, Hyeon-Yong
    • Journal of Microbiology and Biotechnology
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    • v.9 no.3
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    • pp.323-327
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    • 1999
  • It was shown that brain-derived neurotrophic factors (BDNFs) secreted from human neuroblastoma cells can significantly improve the growth of the neurites of PC12 nerve cells. The addition of purified BDNFs elongated the neurites of PC 12 nerve cells two to three times more than the case where the addition was not made. The perfusion rate strongly affected the change of the size of human neuroblastoma cells because the cell size decreased as the perfusion rate increased. This could also influence the productivity of BDNF from the cells. It is also important to note that the BDNF production was decreased when the cell size was reduced. BDNF production rate also decreased at a fast perfusion rate in a smaller cell size. At the relatively fast perfusion rate of 18 ml/h, the ratio of apoptotic to necrotic cells dramatically decreased, which possibly caused the decrease of BDNF production. It has been proven that the secretion of BDNF from human neuroblastoma cells was a partially growth-related process by yielding 6.2$\times l0^{-8}/g$ of BDNF/cell/h of growth related parameter and $0.48{\times}l0^{-9}/g$ of BDNF/cell/h of nongrowth-related parameter in a growth kinetic model. In addition, it was also found that the perfusion rate played a very important role in controlling the cell death mechanism.

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Effects of Berberine on Serum Levels of Catecholamines after Immobilization Stress in Mice (Berberine이 구속 스트레스 부가후 Mouse의 혈중 Catecholamine 함량에 미치는 영향)

  • Shin, Jeong Soo;Lee, Sang Sun;Kim, Eung Il;Shim, Seong Min;Lee, Myung Koo
    • Korean Journal of Clinical Pharmacy
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    • v.7 no.2
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    • pp.81-85
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    • 1997
  • Berberine, a protoberberine isoquinoline alkaloid, showed inhibitory effects on dopamine content in PC12 cells $(53.8\%\;inhibition\;at\;20\;{\mu}M)$. Tyrosine hydroxylase, the rate-limiting enzyme in the catecholamine biosynthetic pathway, was inhibited at $20\;{\mu}M)$ of berberine by $21.8\%$ relative to control. Thus, we hypothesized that the inhibition of tyrosine hydroxylase by berberine might be partially contributed to the decrease in dopamine content in PC12 cells. Furthermore, we investigated the effects of berberine on catecholamine content of serum after immobilization and cold stress in mice. Adult male mice were either subjected to 30 min of restraint or to 2 hr of cold chamber at $4-6^{\circ}C$. Serum norepinephrine, 16.8 pmol/ml, in control mice was increased to 28.8 pmol/ml by immobilization and the stress-induced rise in serum norepinephrine was partially blocked by the treatment of berberine (10 mg/kg, i.p.) once a day for 6 days. Berberine (10 mg/kg/day for 3 days, i.p.) also inhibited the increase in serum norepinephrine by cold stress in mice. These results suggest that berberine may be developed as the promising antistress agent.

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Anti-stress effects of ginseng via down-regulation of tyrosine hydroxylase (TH) and dopamine ${\beta}$-hydroxylase (DBH) gene expression in immobilization-stressed rats and PC12 cells

  • Kim, Yang-Ha;Choi, Eun-Ha;Doo, Mi-Ae;Kim, Joo-Yeon;Kim, Chul-Jin;Kim, Chong-Tai;Kim, In-Hwan
    • Nutrition Research and Practice
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    • v.4 no.4
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    • pp.270-275
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    • 2010
  • Catecholamines are among the first molecules that displayed a kind of response to prolonged or repeated stress. It is well established that long-term stress leads to the induction of catecholamine biosynthetic enzymes such as tyrosine hydroxylase (TH) and dopamine ${\beta}$-hydroxylase (DBH) in adrenal medulla. The aim of the present study was to evaluate the effects of ginseng on TH and DBH mRNA expression. Repeated (2 h daily, 14 days) immobilization stress resulted in a significant increase of TH and DBH mRNA levels in rat adrenal medulla. However, ginseng treatment reversed the stress-induced increase of TH and DBH mRNA expression in the immobilization-stressed rats. Nicotine as a ligand of the nicotinic acetylcholine receptor (nAChR) in adrenal medulla stimulates catecholamine secretion and activates TH and DBH gene expression. Nicotine treatment increased mRNA levels of TH and DBH by 3.3- and 3.1-fold in PC12 cells. The ginseng total saponin exhibited a significant reversal in the nicotine-induced increase of TH and DBH mRNA expression, decreasing the mRNA levels of TH and DBH by 57.2% and 48.9%, respectively in PC12 cells. In conclusion, immobilization stress induced catecholamine biosynthetic enzymes gene expression, while ginseng appeared to restore homeostasis via suppression of TH and DBH gene expression. In part, the regulatory activity in the TH and DBH gene expression of ginseng may account for the anti-stress action produced by ginseng.

Phenolic Profiles of Hardy Kiwifruits and Their Neuroprotective Effects on PC-12 and SH-SY5Y Cells against Oxidative Stress

  • Jeong, Ha-Ram;Kim, Kwan Joong;Lee, Sang Gil;Cho, Hye Sung;Cho, Youn-Sup;Kim, Dae-Ok
    • Journal of Microbiology and Biotechnology
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    • v.30 no.6
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    • pp.912-919
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    • 2020
  • Hardy kiwifruits (Actinidia arguta Planch.) have high amounts of antioxidants, including ascorbic acid (vitamin C) and phenolics. The anti-cholinesterase activity and neuroprotective effects of three different cultivars of hardy kiwifruits, cv. Mansu (A. arguta × A. deliciosa), cv. Haeyeon (A. arguta), and cv. Chiak (A. arguta), on PC-12 and SH-SY5Y cells were evaluated. Extraction of phenolics and vitamin C was carried out using 80% (v/v) aqueous ethanol and metaphosphoric acid assisted with homogenization, respectively. Hardy kiwifruit of cv. Mansu showed higher total phenolic, total flavonoid, and vitamin C contents and antioxidant capacity compared to the other two cultivars of hardy kiwifruits, cv. Haeyeon and cv. Chiak. Analysis of high-performance liquid chromatography results revealed the presence of procyanidin B2, (-)-epicatechin, neochlorogenic acid, cryptochlorogenic acid, rutin, hyperoside, isoquercitrin, and astragalin in hardy kiwifruits. The three cultivars of hardy kiwifruits had a wide range of vitamin C content of 55.2-130.0 mg/100 g fresh weight. All three cultivars of hardy kiwifruits had protective effects on neuronal PC-12 and SH-SY5Y cells exposed to hydrogen peroxide by increasing cell viability and reducing intracellular oxidative stress. Furthermore, the hardy kiwifruits inhibited acetylcholinesterase and butyrylcholinesterase. Collectively, these results suggest that hardy kiwifruits rich in antioxidants like phenolics and vitamin C have good potential as functional materials in neuroprotective applications.

Effects of Combined Treatments of Lithium and Valproate on the Phosphorylation of ERK1/2 and Transcriptional Activity of ELK1 and C-FOS in PC12 Cells (리튬 및 발프로에이트 병용 처치가 PC12 세포에서 ERK1/2 인산화와 ELK1 및 C-FOS 전사활성에 미치는 영향)

  • Cha, Seung Keun;Kim, Se Hyun;Ha, Kyooseob;Shin, Soon Young;Kang, Ung Gu
    • Korean Journal of Biological Psychiatry
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    • v.20 no.4
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    • pp.159-165
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    • 2013
  • Objectives Mechanisms of clinical synergistic effects, induced by co-treatments of lithium and valproate, are unclear. Extracellular signal-regulated kinase (ERK) has been suggested to play important roles in mechanisms of the action of mood stabilizers. In this study, effects of co-treatments of lithium and valproate on the ERK1/2 signal pathway and its down-stream transcription factors, ELK1 and C-FOS, were investigated in vitro. Methods PC12 cells, human pheochromocytoma cells, were treated with lithium chloride (30 mM), valproate (1 mM) or lithium chloride + valproate. The phosphorylation of ERK1/2 was analyzed with immunoblot analysis. Transcriptional activities of ELK1 and C-FOS were analyzed with reporter gene assay. Results Single treatment of lithium and valproate increased the phosphorylation of ERK and transcriptional activities of ELK1 and C-FOS, respectively. Combined treatments of lithium and valproate induced more robust increase in the phosphorylation of ERK1/2 and transcriptional activities of ELK1 and C-FOS, compared to those in response to single treatment of lithium or valproate. Conclusions Co-treatments of lithium and valproate induced synergistic increase in the phosphorylation of ERK1/2 and transcriptional activities of its down-stream transcription factors, ELK1 and C-FOS, compared to effects of single treatment. The findings might suggest potentiating effects of lithium and valproate augmentation treatment strategy.

Deastringent Peel Extracts of Persimmon (Diospyros kaki Thunb. cv. Cheongdo-Bansi) Protect Neuronal PC-12 and SH-SY5Y Cells against Oxidative Stress

  • Jeong, Da-Wool;Cho, Chi Heung;Lee, Jong Suk;Lee, Seung Hwan;Kim, Taewan;Kim, Dae-Ok
    • Journal of Microbiology and Biotechnology
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    • v.28 no.7
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    • pp.1094-1104
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    • 2018
  • The peel of astringent persimmon (Diospyros kaki Thunb. cv. Cheongdo-Bansi) is a by-product of dried persimmon (gotgam). We investigated if deastringent peel extracts of persimmon cv. Cheongdo-Bansi had antioxidative and neuroprotective properties. Two different extracts were prepared: thermally and nonthermally treated persimmon peel extracts (TPE and NTPE, respectively). Both TPE and NTPE were fractionated sequentially in n-hexane, chloroform, ethyl acetate, n-butanol, and water. The TPE and NTPE ethyl acetate fractions had the highest total phenolic and flavonoid contents as well as antioxidant capacities among all the fractions. Pretreatment of neuronal PC-12 and SH-SY5Y cells with the TPE and NTPE ethyl acetate fractions increased cell viability after exposure to oxidative stress. The ethyl acetate fraction of TPE attenuated oxidative stress inside both PC-12 and SH-SY5Y cells more effectively than that of NTPE. Furthermore, the TPE and NTPE ethyl acetate fractions inhibited acetylcholinesterase and butyrylcholinesterase. Analysis of ultra-high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry results revealed gallic acid, kaempferol, kaempferol-3-O-galactoside, kaempferol-3-O-glucoside, quercetin, quercetin3-O-galactoside, quercetin-3-O-galactoside-2'-O-gallate, and quercetin-3-O-glucoside as the major phenolics of the TPE and NTPE ethyl acetate fractions. Taken together, these results suggest that the ethyl acetate fraction of deastringent persimmon peel is rich in antioxidants and has potential as a functional food to reduce oxidative stress.

Altered APP Carboxyl-Terminal Processing Under Ferrous Iron Treatment in PC12 Cells

  • Kim, Chi Hyun;Yoo, Yeong-Min
    • The Korean Journal of Physiology and Pharmacology
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    • v.17 no.3
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    • pp.189-195
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    • 2013
  • Amyloid-${\beta}$ peptide ($A{\beta}$), generated by proteolytic cleavage of the amyloid precursor protein (APP), plays a pivotal role in the pathogenesis of Alzheimer's disease (AD). The key step in the generation of $A{\beta}$ is cleavage of APP by beta-site APP-cleaving enzyme 1 (BACE1). Levels of BACE1 are increased in vulnerable regions of the AD brain, but the underlying mechanism is unknown. In the present study, we reported the effects of ferrous ions at subtoxic concentrations on the mRNA levels of BACE1 and a-disintegrin-and-metalloproteinase 10 (ADAM10) in PC12 cells and the cell responses to ferrous ions. The cell survival in PC12 cells significantly decreased with 0 to 0.3 mM $FeCl_2$, with 0.6 mM $FeCl_2$ treatment resulting in significant reductions by about 75%. 4,6-diamidino-2-phenylindole (DAPI) staining showed that the nuclei appeared fragmented in 0.2 and 0.3 mM $FeCl_2$. APP-${\alpha}$-carboxyl terminal fragment (APP-${\alpha}$-CTF) associations with ADAM10 and APP-${\beta}$-CTF with BACE1 were increased. Levels of ADAM10 and BACE1 mRNA increased in response to the concentrations of 0.25 mM, respectively. In addition, p-ERK and p-Bad (S112, S155) expressions were increased, suggesting that APP-CTF formation is related to ADAM10/ BACE1 expression. Levels of Bcl-2 protein were increased, but significant changes were not observed in the expression of Bax. These data suggest that ion-induced enhanced expression of AMDA10/BACE1 could be one of the causes for APP-${\alpha}/{\beta}$-CTF activation.

Protective effect of ethanolic extract of antler-shaped Ganoderma lingzhi against oxidative stress in PC12 neuronal cell line (PC12 신경세포주에서 녹각영지버섯 주정 추출물의 산화 스트레스 개선 효과)

  • Kim, Hyung Don;Lee, Eun Young;Park, Jeong-Yong;Seo, Kyung Hye;Lee, Kang-Hyo;Choi, Jehun;Han, Jae-Gu;Cho, Jae-Han;Lee, Seung Eun
    • Journal of Mushroom
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    • v.16 no.3
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    • pp.213-217
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    • 2018
  • This study was carried out to identify medicinal mushrooms with protective effects against oxidative stress in PC12 neuronal cell line, followed by evaluation of their antioxidant property. Extracts of medicinal mushrooms, including Ganoderma lucidum extract (GLE), antler-shaped Ganoderma lingzhi extract (AGLE), Hericium erinaceus extract (HEE), and Sanghuangporus baumii extract (SBE), were screened for cytotoxicity using MTT assay. None of the extracts up to $10{\mu}g/ml$ concentration affected cell viability. These extracts were further checked for their protective effect against oxidative stress-induced reactive oxygen species (ROS) production. Exposure to $50{\mu}M$ $H_2O_2$ induced ROS generation in PC12 cells, which was inhibited only by treatment with AGLE. In addition, inhibition of $H_2O_2-induced$ ROS generation by AGLE was found to be in a dose-dependent manner (2.5, 5, and $10{\mu}g/ml$). Microscopic examination of DCF fluorescence for detection of ROS showed a similar pattern. Further, antioxidant activity of AGLE was determined by ABTS radical cation assay, and its $IC_{50}$ was found to be $46.90{\pm}0.31{\mu}g/ml$. Taken together, these results suggest that AGLE may help to alleviate oxidative stress in PC12 neuronal cells.