• The Journal of Korean Obstetrics and Gynecology
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• v.22 no.2
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• pp.119-131
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• 2009

Purpose: The depression accompanied with menopuase shows the relation with the dopamine secretion. These studies were undertaken to evaluate the anti- oxidative and neuroprotective effects of Bunsimgieum(BSGE) on dopaminergic neurons. Methods: To estimate the antioxidant effects, we carried out 1.1-diphenyl-2- picrylhydrazyl (DPPH) free radical scavenging assay, 2,2'-azinobis-(3-ethylbenzothiazoline -6-sulfonic acid (ABTS) radical cation decolorization assay, and measurement of total polyphenolic content. To evaluate neuroprotective effect of BSGE in vitro, We performed thiazolyl blue tetrazolium bromide (MTT) assay, reactive oxygen species (ROS) creation in SH-SY5Y. Tyrosine hydroxylase (TH) immunocytochemistry, nitric oxide (NO) assay, and TNF-${\alpha}$ assay in primary rat mesencephalic dopaminergic neurons. Results: The DPPH free radical and the ABTS radical cation inhibition activities were increased at a dose dependent manner. Total polyphenolic content was 0.83%. In SH-SY5Y culture, BSGE significantly increased the decreased cell viability by 6-OHDA at the concentrations of 10${\mu}$g/m${\ell}$ in pre-treatment group, 0.1-200${\mu}$g/m${\ell}$ in post-treatment group. The production of ROS induced by 6-OHDA was significantly inhibited in BSGE treated group. In mesencephalic dopaminergic cell culture, the BSGE group reduced the dopaminergic cell loss against 6-OHDA toxicity and the production of No and TNF-${\alpha}$ at the concentration of 5${\mu}$g/m${\ell}$. Conclusion: These results shows that BSGE has antioxidant and neuroprotective effects in the dopaminergic cells through decreasing the production of ROS, NO and TNF-${\alpha}$ which can cause many neurodegenerative changes in brain cell. We suggest that BSGE could be useful for the treatment of postmenopausal depression related with the decrease of dopamine.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.1
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• pp.1-11
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• 2016

This study was conducted to investigate effect of the acetylcholinesterase inhibitor activity, the protective effect of the extract on SH-SY5Y cell death by $H_2O_2$, the memory improvement from scopolamine-induced rat. Moreover, the antioxidant activity of isorhamnetin from the dropwort (Oenanthe javanica) was investigated. Acetylcholinesterase inhibitor activity was highest (28.59%) in Hwasun O. javanica extract (H-OJE). H-OJE and Naju O. javanica extract (N-OJE) were not significantly different. SH-SY5Y cell death deceased to 37.23% and 36.68% for H-OJE and N-OJE, respectively, following treatment with the extracts. O. javanica extracts showed a protective effect against $H_2O_2$-induced neurotoxicity. Treatment with O. javanica extracts slightly improved scopolamine-induced (1 mg/kg, i.p.) memory impairment in rats. H-OJE contained the highest total phenolic and flavonoid contents of 117 mg/g and 30 mg gallic acid equivalents/g, respectively, and had a DPPH radical scavenging activity ($SC_{50}$) of $113.8{\mu}g/mL$ and ABTS radical scavenging activity of $48.2{\mu}g/mL$, which was higher than the other extracts. The thiobarbituric acid reactive substances value was highest (50.2%) in H-OJE. Antioxidant activity differed significantly among dropwort extracts. Isorhamnetin was known as one of the flavonoid and for having neuroprotective effect. So we analyzed acid-hydrolyzed O. javanica extract HPLC. The results were that peak at 14 min and spectrum of the extracts was consistent with standard solution. The results of LC/MS/MS analysis were that the extract and standard solution were confirmed total ion chromatogram at identical time, precursor ion was 317 $[M-H]^+$ m/z, product ion was 302 $[M-H]^+$ m/z. Overall, the results showed that the dropwort extract led to memory improvement and had antioxidant activity. Based on these finding, further research to investigate the production of ethanol extract of dropwort as a processed food is warranted.

• BMB Reports
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• v.45 no.9
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• pp.532-537
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• 2012

P18, a member of the INK4 family of cyclin-dependent kinase inhibitors, is a tumor suppressor protein and plays a key cell survival role in a variety of human cancers. Under pathophysiological conditions, the INK4 group proteins participate in novel biological functions associated with neuronal diseases and oxidative stress. Parkinson's disease (PD) is characterized by loss of dopaminergic neurons, and oxidative stress is important in its pathogenesis. Therefore, we examined the effects of PEP-1-p18 on oxidative stress-induced SH-SY5Y cells and in a PD mouse model. The transduced PEP-1-p18 markedly inhibited 1-methyl-4-phenyl pyridinium-induced SH-SY5Y cell death by inhibiting Bax expression levels and DNA fragmentation. Additionally, PEP-1-p18 prevented dopaminergic neuronal cell death in the substantia nigra of a 1-methyl-4-phenyl-1,2,3,6,-tetrahydropyridine-induced PD mouse model. These results indicate that PEP-1-p18 may be a useful therapeutic agent against various diseases and is a potential tool for treating PD.

• BMB Reports
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• v.42 no.7
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• pp.444-449
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• 2009

Different neurodegenerative disorders like prion disease, is caused by protein misfolding conformers. Reverse-transfected cytosolic prion protein (PrP) and PrP expressed in the cytosol have been shown to be neurotoxic. To investigate the possible mechanism of neurotoxicity due to accumulation of PrP in cytosol, a PrP mutant lacking the signal and GPI (CytoPrP) was introduced into the SH-SY5Y cell. MTT and trypan blue assays indicated that the viability of cells expressing CytoPrP was remarkably reduced after treatment of MG-132. Obvious apoptosis phenomena were detected in the cells accumulated with CytoPrP, including loss of mitochondrial transmembrane potential, increase of caspase-3 activity, more annexin V/PI-double positive-stained cells and reduced Bcl-2 level. Moreover, DNA fragmentation and TUNEL assays also revealed clear evidences of late apoptosis in the cells accumulated CytoPrP. These data suggest that the accumulation of CytoPrP in cytoplasm may trigger cell apoptosis, in which mitochondrial relative apoptosis pathway seems to play critical role.

• Journal of Oriental Neuropsychiatry
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• v.21 no.2
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• pp.125-140
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• 2010

Objectives : The purpose of this study is to examine from various angles the protective effect of Gastrodia elata Blume (GEB) against nerve cell death induced by $\beta$-amyloid by using the cell line SH-SY5Y, which is commonly utilized for toxicity testing in nerve cells, and to find out its mechanism of action. Methods : To begin with, as a result of assessing the rate of cell survival by employing MTT reduction assay, the treatment with $\beta$-amyloid at different concentrations caused cytotoxicity, which was inhibited by preprocessing GEB extract. In addition, after $\beta$-amyloid was processed with the cell SH-SY5Y, apoptosis progressed, which was reduced effectively by processing GEB extract. Results : When cytotoxicity was caused by using hydrogen peroxide, a representative ROS, in order to examine the antioxidant effect of GEB, its protective effect was also observed. Apart from ROS, reactive nitrogen species (RNS) are also known to play a crucial role in nerve cell death. The treatment with the NO donor SNAP increased the production of nitric oxide and the expression of iNOS, which was also inhibited by GEB extract. Meanwhile, as an attempt to find out the mechanism of action explaining the antioxidant effect, the intracellular antioxidant enzyme expressions were measured by RT-PCR, which showed that GEB extract increased the expressions of heme oxygenase-1, GAPDH and $\gamma$-glutamate cysteine ligase. Lastly, GEB extract had a protective effect against impaired memory induced by scopolamine in animal models (in vivo). Conclusions : These findings indicate that GEB has a protective effect against the death of cranial nerve cells, suggesting possibilities for the prevention and treatment of AD.

• BMB Reports
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• v.48 no.7
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• pp.395-400
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• 2015

Parkinson's disease (PD) is a neurodegenerative disability caused by a decrease of dopaminergic neurons in the substantia nigra (SN). Although the etiology of PD is not clear, oxidative stress is believed to lead to PD. Catalase is antioxidant enzyme which plays an active role in cells as a reactive oxygen species (ROS) scavenger. Thus, we investigated whether PEP-1-Catalase protects against 1-methyl-4-phenylpyridinium (MPP⁺) induced SH-SY5Y neuronal cell death and in a 1-methyl-4-phenyl-1,2,3,6-trtrahydropyridine (MPTP) induced PD animal model. PEP-1-Catalase transduced into SH-SY5Y cells significantly protecting them against MPP⁺-induced death by decreasing ROS and regulating cellular survival signals including Akt, Bax, Bcl-2, and p38. Immunohistochemical analysis showed that transduced PEP-1-Catalase markedly protected against neuronal cell death in the SN in the PD animal model. Our results indicate that PEP-1-Catalase may have potential as a therapeutic agent for PD and other oxidative stress related diseases. [BMB Reports 2015; 48(7): 395-400]

• BMB Reports
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• v.37 no.2
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• pp.185-191
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• 2004

Tissue transglutaminase (tTGase) regulates various biological processes, including extracellular matrix organization, cellular differentiation, and apoptosis. Here we report the protective role of tTGase in the cell death that is induced by the tumor necrosis factor $\alpha$ (TNF-$\alpha$) and ceramide, a product of the TNF-$\alpha$ signaling pathway, in human neuroblastoma SH-SY5Y cells. Treatment with retinoic acid (RA) induced the differentiation of the neuroblastoma cells with the formation of extended neurites. Immunostaining and Western blot analysis showed the tTGase expression by RA treatment. TNF-$\alpha$ or $C_2$ ceramide, a cell permeable ceramide analog, induced cell death in normal cells, but cell death was largely inhibited by the RA treatment. The inhibition of tTGase by the tTGase inhibitors, monodansylcadaverine and cystamine, eliminated the protective role of RA-treatment in the cell death that is caused by TNF-$\alpha$ or $C_2$-ceramide. In addition, the co-treatment of TNF-$\alpha$ and cycloheximide ecreased the protein level of tTGase and cell viability in the RA-treated cells, supporting the role of tTGase in the protection of cell death. DNA fragmentation was also induced by the co-treatment of TNF-$\alpha$ and cycloheximide. These results suggest that tTGase expressed by RA treatment plays an important role in the protection of cell death caused by TNF-$\alpha$ and ceramide.

• Food Science and Preservation
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• v.14 no.4
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• pp.425-430
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• 2007

To investigate the potential therapeutic value of a plant extract against amyloid ${\beta}-peptide-induced$ cell damage, we first screened extracts of 250 herbs, and finally selected a water extract of Spinacia oleracea for further study. This extractshowed the potential to inhibit the reactions of oxidants. We measured the angiotensin-converting-enzyme (ACE) inhibitory activity of the extract, and assessed the ability of the extract to protect neuronal cells from chemical-induced cell death. SH-SY5Y neuroblastoma cells were used in this assay. The extract exerted protective effects on $H_2O_2-induced$ cell death, when $H_2O_2$ was used at 100 M, 200 M, and 500 M (protection of 87%, 73%, and 58%, respectively). When 50 M of amyloid ${\beta}-peptide$ was added to the test cells, however, the extract had no protective effect on cell death. The extract inhibited ACE activity in a dose-dependent manner, and exhibited potent protection against the deleterious effects of $H_2O_2$. In sum, these results suggest that a water extract of Spinacia oleracea has the potential to afford protection against chemical-induced neuronal cell death, and the extract may be useful in the treatment of neurodegenerative diseases. The precise molecular mechanism of neuroprotection is under investigation.

• Molecules and Cells
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• v.39 no.7
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• pp.543-549
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• 2016

MicroRNAs (miRNAs) have been reported to be involved in many neurodegenerative diseases. The present study focused on the role of hsa-miR-144-3p in one of the neuro-degenerative diseases, Parkinson's disease (PD). Our study showed a remarkable down-regulation of miR-144-3p expression in 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP)-treated SH-SY5Y cells. MiR-144-3p was then overexpressed and silenced in human SH-SY5Y cells by miRNA-mimics and miRNA-inhibitor transfections, respectively. Furthermore, ${\beta}$-amyloid precursor protein (APP) was identified as a target gene of miR-144-3p via a luciferase reporter assay. We found that miR-144-3p overexpression significantly inhibited the protein expression of APP. Since mitochondrial dysfunction has been shown to be one of the major pathological events in PD, we also focused on the role of miR-144-3p and APP in regulating mitochondrial functions. Our study demonstrated that up-regulation of miR-144-3p increased expression of the key genes involved in maintaining mitochondrial function, including peroxisome proliferator-activated receptor ${\gamma}$ coactivator-$1{\alpha}$(PGC-$1{\alpha}$), nuclear respiratory factor 1 (NRF-1) and mitochondrial transcription factor A (TFAM). Moreover, there was also a significant increase in cellular ATP, cell viability and the relative copy number of mtDNA in the presence of miR-144-3p overexpression. In contrast, miR-144-3p silencing showed opposite effects. We also found that APP overexpression significantly decreased ATP level, cell viability, the relative copy number of mtDNA and the expression of these three genes, which reversed the effects of miR-144-3p overexpression. Taken together, these results show that miR-144-3p plays an important role in maintaining mitochondrial function, and its target gene APP is also involved in this process.

• The Journal of Korean Obstetrics and Gynecology
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• v.22 no.1
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• pp.1-14
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• 2009

Purpose: Oxidative stress was thought to play a critical role in neurodegenerative disease. Many in vivo and in vitro reports explained the possible pathway of human aging. But in therapeutic aspects, there was no clear answers to prevent aging associated with neural diseases. In this study, we investigated the antioxidant and neuroprotective effects of the Insamyangyung-tang (IYT). Methods: To estimate the antioxidant effects, we carried out 1.1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay, 2,2'-azinobis-(3- ethylbenzothiazoline-6- sulfonic acid (ABTS) radical cation decolorization assay, and measurement of total polyphenolic content. To evaluate neuroprotective effect of IYT in vitro. We performed thiazolyl blue tetrazolium bromide (MTT) assay, reactive oxygen species (ROS) creation in SH-SY5Y. Tyrosine hydroxylase (TH) immunocytochemistry, nitric oxide (NO) assay, and TNF-${\alpha}$ assay in primary rat mesencephalic dopaminergic neurons. Results: The $IC_{50}$ values were $571.6{\mu}g/m{\ell}$ and $202.3{\mu}g/m{\ell}$ in DPPH and ABTS assay respectively. Total polyphenolic content was 1.05%. In SH-SY5Y culture, IYT significantly increased the decreased cell viability by 6-OHDA at the concentrations of $10{\mu}g/m{\ell}$ in pre-treatment group, $10-100{\mu}g/m{\ell}$ in post-treatment group, and $100{\mu}g/m{\ell}$ in co-treatment group. The production of ROS induced by 6-OHDA was significantly inhibited in IYT treated group. In mesencephalic dopaminergic cell culture, the IYT group reduced the dopaminergic cell loss against 6-OHDA toxicity and the production of No and TNF-${\alpha}$ at the concentration of $0.2{\mu}g/m{\ell}$. Conclusion: These results showed that IYT has antioxidant and neuroprotectctive effects in the dopaminergic cells through decreasing the production of ROS, NO and TNF-${\alpha}$ which can cause many neurodegenerative changes in brain cell.