• The Korea Journal of Herbology
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• v.24 no.2
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• pp.29-37
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• 2009

Objectives: The goal of this study was to investigate the antidepressive effect of Bupleuri Radix (BR). Methods : The forced swimming test (FST) was performed. Also the expression of corticotropin releasing factor (CRF), c-Fos and tyrosine hydroxylase (TH) was measured immunohistochemically at paraventricular nucleus (PVN) and locus coeruleus (LC). Concentration of adrenocorticotrophic hormone (ACTH) was measured in plasma by ELISA method. Results : The immobility in BR400 Group was significantly decreased in comparison with the control group (p

• The Korea Journal of Herbology
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• v.24 no.2
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• pp.87-92
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• 2009

Objectives : This study was performed to evaluate the neuroprotective effect of water extracts from Nelumbinis semen (NSW) in dopaminergic cells. Methods : We performed 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay, 2,2-azinobis3-ethyl-benzothiazoline-6-sulfonic acid (ABTS) cation scavenging assay, and determination of total polyphenolic content to examine the antioxidant effects of NSW. We also evaluated the neuroprotective effects against 6-hydroxydopamine (6-OHDA)-induced toxicity using 3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyl-tetrazolium bromide assay (MIT) assay, trypan blue cytotoxicity assay, and nitric oxide assay in SH-SY5Y cells and tyrosine hydroxylase (TH) immunohistochemistry in primary rat dopaminergic neurons. Results : NSW showed $IC_{50}$ values of 184.80 and 92.90 ${\mu}$g/mL in DPPH and in ABTS assays, respectively. NSW showed 1.05% of total polyphenol contents. NSW showed protective effect against 6-0HDA-induced neurotoxicity whereas no influence on cell viability at the concentration of 1${\sim}$50 ${\mu}$g/mL. NSW reduced NO generation while 6-OHDA produced it. Moreover, it protected rat dopaminergic neurons against 6-0HDA at a dose of 1 ${\mu}$g/mL. Conclusions : These results indicated that NSW has neuroprotective effect against 6-0HDA-induced neurotoxicity through antioxidant activity in dopaminergic cell culture.

• Korean Journal of Acupuncture
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• v.39 no.1
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• pp.8-15
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• 2022

Objectives : Saam acupuncture is one of the indigenous therapeutic modalities in traditional Korean medicine. In this study, the neuroprotective effect of Saam acupuncture of kidney tonification was investigated using 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated mice. Methods : Twelve-week-old male C57BL/6 mice were intraperitoneally administered with 30 mg/kg of MPTP at 24-h intervals for 5 days and acupuncture stimulation at LU8, KI7, SP3 and KI3 was performed once a day for 12 days from the first MPTP injection. The pole test and the rotarod test were performed to evaluate motor function, and dopaminergic neuronal survival in the substantia nigra (SN) and striatum was evaluated using tyrosine-hydroxylase immunohistochemistry. Results : MPTP administration caused behavioral impairment and dopaminergic neuronal death in the nigrostriatal pathway. Whereas the Saam acupuncture treatment alleviated the MPTP-induced motor dysfunction and dopaminergic neuronal death in the SN and striatum. Conclusions : Saam acupuncture of kidney tonification can alleviate the MPTP-induced motor dysfunction and dopaminergic neuronal death in the nigrostriatal pathway, suggesting a possible role for acupuncture in the treatment of Parkinson's disease.

• Applied Microscopy
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• v.30 no.2
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• pp.193-204
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• 2000

Nowadays, mongolian gerbil is widely utilized in the research of brain and water deprivation because of congenitally incomplete Willis' circle, audiogenic seizure in low noise, and special cholesterol metabolism without water absorption for a long time. In this study, we intended to identify the time lapse changes in the general morphoogy and ultrastructure of the catecholaminergic neurons of mongolian gerbil brain in after long-term water deprivation. Fifteen mongolian gerbils were divided into 3 groups (5, 10, and 20-day water deprivation groups), each with 5 mongolian gerbils. Additional 5 mongolian gerbils which received water without limitation were used as a control. The brain sections were immunostained using tyrosine hysroxylase (TH), $ dopamine-\beta-hydroxylase$ (DBH), and phenylethanolamine-N-methyltrasferase (PMNT) antibodies. And immunoreactive cells were observed by electromicroscopy for the ultrastructural changes . The TH-immunoreactive (TH-IR) nerve cells were observed in the para- and peri-ventricular nucleus of the 3 rd ventricle in the hypothalamus and the substantia nigra. The number of TH-IR neurons in these areas was decreased from the 5th day of the water deprivation to the 10 th day and reincreased until 20 th day water deprivation. The shape and density of the dopamine-secreting cells identified by immunohistochemistry showed changes in the continuous water deprivation. Electron microscopy revealed a round nucleus in the neurons of control group but 5-day water deprivation group showed a dense and irregularly shaped nucleus. Also in the 5-day water-deprived group, mitochondria was decreased in number and junctins were disappered. Endoplasmic reticulum, Golgi complex did not show changes after water-deprivation. In this results, we can conclude that dopamine are involved in the water metabolism in mongolian gerbil, and mongolian gerbil could be used as an animal model for the researches of water deprivation.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.34 no.4
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• pp.287-301
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• 2008

In order to develop a new depigmenting agent, extracts were obtained from 60 native plants and their antimelanogenic activities were screened by evaluating the inhibitory effect on tyrosinase which is a major enzyme responsibles for the melanin synthesis. The extracts of Trichosanthes kirilowii fruits, Phyllostachys bambusoides inner films (BIF), Clerodendrum trichotomum leaves, and Acer okamotoanum leaves showed relatively high inhibitory effect on tyrosinase and their $IC_{50}$ values were $50{\sim}100{\mu}g/mL$. The extract of BIF inhibited melanin synthesis of B16F10 melanoma cells by 52%, which was the highest among those of various extracts. Furthermore, the effect of BIF extract is 10% higher than that of arbutin (42%), a popular depigmenting agent in Korea. Ten compounds having antimelanogenic activity were isolated from the BIF extract by solvent extraction and chromatography. These compounds were identified as phenolic derivatives: SM701, SM702, SM703, and BPR211 were hydroquinone derivatives; SM707 a gallic acid derivative; SM704, SM705, SM706, SM708 and SM709 ferulic acid derivatives. The free radical scavenging activities of these compounds were measured and compared to those of hydroquinone and vitamin C. The $SC_{50}$ values scavenging 50% DPPH of SM702 and SM709 were $60{\sim}70{\mu}M$ similar to that of hydroquinone and those of SM701 and SM708 were $30{\sim}40{\mu}M$ slightly lower than that of vitamin C. These results suggest the presence of components having high antioxidant activity in the BIF extract. The SM709, identified as 1,2-O-diferulylglycerol, inhibited the activities of tyrosine hydroxylase and dopa oxidase by 18 and 60%, respectively. The SM709 also inhibited the melanin synthesis of B16F10 melanoma cells by 62% and this was the highest antimelanogenic activity among those obtained from the various purified compounds. Therefore, antimelanogenic activity of the BIF extract was concluded to be due to both inhibition of DOPA oxidase and antioxidant activity.

• Science of Emotion and Sensibility
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• v.10 no.3
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• pp.411-418
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• 2007

Exposure to low frequency noise(LFN) can lead to vibroacoustic diseases(VADs), which include a systemic disease with lesions in a broad spectrum of organs and a psychiatric condition. It is known that VAD is an established risk factor for the development of many psychological conditions in humans and rodents, including major depression and anxiety disorder. The present study investigated the effects of LFN on neuronal stress responses in the rat brain. The neuronal expression of the proto-oncogene c-fos in the paraventricular nucleus(PVN) of the hypothalamus and tyrosine hydroxylase(TH) in the LC was observed. The immunocytochemical detection of the Fos protein and TH has been used as a marker of neuronal activation in response to stress. In addition, corticosterone concentration was evaluated by using an enzyme-linked immunosorbent assay(ELISA). The LFN groups were exposed to 32.5Hz and 125Hz of noise(4hr/day for 2days). The numbers of c-fos and TH-immunoreactive cells in the PVN and LC were significantly increased in the LFN groups(32.5Hz and 125Hz) compared to the normal group. Corticosterone concentration in plasma was also increased in LFN groups. The present results demonstrated that exposure with LFN produced a pronounced increase in expression of c-Fos and TH in stress-relevant brain areas. These results suggest that the neural characteristics involved in LFN are similar to those activated by typical processive stressors. These results also suggest that the central and peripheral activations by LFN may be related to LFN-related negative behavioral dysfunctions such as VADs.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.26 no.1
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• pp.149-162
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• 2000

Coenzyme Q10 is found in all tissues including skin and it is the well-known coenzyme for mitochondrial enzymes. The electron and proton transfer functions of the quinone ring are of fundamental importance for the oxidative phosphorylation pathway to generate energy in the cells. Coenzyme Q10 has been studied as a potent antioxidant molecule in the skin. It is involved in the skin's response to UVR irradiation. The concentration of this antioxidant in UVR exposed skin is higher than in non-exposed skin. However, recent studies have also shown that coenzyme Q10 is one of the first antioxidants to be depleted when skin is UVR-irradiated. This indicates that coenzyme Q10 is primarily involved in defense mechanisms of the skin. Therefore, we questioned whether coenzyme Q10 shows reulatory effect of melanogenesis. Here we report that coenzyme Q10 inhibits melanin neosynthesis of normal human melanocytes grown in culture, and lightens UVB-induced hyperpigmentation of the guinea pig skin in vivo. We treated human melanocytes with 0.05mM to 0.5mM of coenzyme Q10 for a total of two days. This inhibited melanin neosynthesis of cultured human melanocytes dose-dependently. The inhibitory effect of coenzyme Q10 was as effective as kojic acid or vitamin C on cultured human melanocytes. CoQ10 didn't have direct inhibitory effect on tyrosinase activity in in vitro tyrosine hydroxylase activity To further clarify the effect of coenzyme Q10 on the melanogenesis, we established UVB-induced hyperpigmentation on the shaved backs of brownish guinea pigs. The UVB intensity was 500mJ/$\textrm{cm}^2$ and the total energy dose was 1,500 mJ/$\textrm{cm}^2$. The animals were exposed to UVB radiation one times a week for three consecutive weeks. Coenzyme Q10, kojic acid, Arbutin, vitamin C(1% in vehicle) or vehicle alone as a control were then topically applied daily to the hyperpigmented areas twelve times per week far four successive weeks. The lightening effect was evaluated by visual scoring, chromameter and immunohistochemistry. Coenzyme Q10 had lightening effect on the UVB-induced hyperpigmentation without any other side effects, whereas another compounds showed weak lightening efficacies. Therefore, these results suggest that coenzyme Q10 may be useful for solving physiological hyperpigmenting problems for cosmetic purposes.

• Development and Reproduction
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• v.13 no.4
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• pp.257-264
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• 2009

A neurotoxin, 6-hydroxydopamine (6-OHDA) has been widely used to create animal model for Parkinson's disease (PD) due to its specific toxicity against dopaminergic (DA) neurons. Since DA signals modulate a broad spectrum of CNS physiology, one can expect profound alterations in neuroendocrine activities of both PD patients and 6-OHDA treated animals. Limited applications of 6-OHDA injection model, however, have been made on the studies of hypothalamuspituitary neuroendocrine circuits. The present study was performed to examine whether blockade of brain catecholamine (CA) biosynthesis with 6-OHDA can make any alteration in the transcriptional activities of hypothalamus-pituitary hormone genes in adult male rats. Three-month-old male rats (SD strain) were received 6-OHDA ($200{\mu}g$ in $10{\mu}\ell$ of saline/animal) by intracerebroventricular (icv) injection, and sacrificed after two weeks. To determine the mRNA levels of hypothalamuspituitary hormone genes, total RNAs were extracted and applied to the semi-quantitative RT-PCRs. The mRNA levels of tyrosine hydroxylase (TH), the rate-limiting enzyme for the catecholamine biosynthesis, were significantly lower than those from the control group (control:6-OHDA=1:0.72${\pm}$0.02AU, p<0.001), confirming the efficacy of 6-OHDA injection. The mRNA levels of gonadotropin-releasing hormone (GnRH) and corticotropin releasing hormone (CRH) in the hypothalami from 6-OHDA group were significantly lower than those from the control group (GnRH, control:6-OHDA=1:0.39${\pm}$0.03AU, p<0.001; CRH, control:6-OHDA=1:0.76${\pm}$0.07AU, p<0.01). There were significant decreases in the mRNA levels of common alpha subunit of glycoprotein homones (Cg$\alpha$), LH beta subunit (LH-$\beta$), and FSH beta subunit (FSH-$\beta$) in pituitaries from 6-OHDA group compared to control values (Cg$\alpha$, control:6-OHDA=1:0.81${\pm}$0.02AU, p<0.001; LH-$\beta$, control:6-OHDA=1:0.68${\pm}$0.04AU, p<0.001; FSH-$\beta$, control:6-OHDA=1:0.84${\pm}$0.05AU, p<0.001). Similarly, the level of adrenocorticotrophic hormone (ACTH) transcripts from 6-OHDA group was significantly lower than that from the control group (control: 6-OHDA=1:0.86${\pm}$0.04AU, p<0.01). The present study demonstrated that centrally injected DA neurotoxin could downregulate the transcriptional activities of the two hypothalamus-pituitary neuroendocrine circuits, i.e., GnRH-gonadotropins and CRH-ACTH systems. These results suggested that hypothalamic CA input might affect on the activities of gonad and adrenal through modulation of hypothalamus-pituitary function, providing plausible explanation for frequent occurrence of sexual dysfunction and poor stress-response in PD patients.

• Clinical and Experimental Reproductive Medicine
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• v.31 no.1
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• pp.19-27
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• 2004

Objective: This study was to examine the in vitro neural cell differentiation patterns of human embryonic stem (hES) cells following treatment of various neurotrophic factors [basic fibroblast growth factor (bFGF), retinoic acid (RA), brain derived neurotrophic factor (BDNF) and transforming growth factor (TGF)-$\alpha$], particulary in dopaminergic neuron formation. Methods: The hES cells were induced to differentiate by bFGF and RA. Group I) In bFGF induction method, embryoid bodies (EBs, for 4 days) derived from hES were plated onto gelatin dish, selected for 8 days in ITSFn medium and expanded at the presence of bFGF (10 ng/ml) for another 6 days followed by a final differentiation in N2 medium for 7, 14 and 21 days. Group II) For RA induction, EBs were exposed of RA ($10^{-6}M$) for 4 days and allowed to differentiate in N2 medium for 7, 14 and 21 days. Group III) To examine the effects of additional neurotrophic factors, bFGF or RA induced cells were exposed to either BDNF (10 ng/ml) or TGF-$\alpha$ (10 ng/ml) during the 21 days of final differentiation. Neuron differentiation and dopamine secretion were examined by indirect immunocytochemistry and HPLC, respectively. Results: The bFGF or RA treated hES cells were resulted in similar neural cell differentiation patterns at the terminal differentiation stage, specifically, 75% neurons and 11% glial cells. Additionally, treatment of hES cells with BDNF or TGF-$\alpha$ during the terminal differentiation stage led to significantly increased tyrosine hydroxylase (TH) expression of a dopaminergic neuron marker, compared to control (p<0.05). In contrast, no effect was observed on the rate of mature neuron (NF-200) or glutamic acid decarboxylase-positive neurons. Immunocytochemistry and HPLC analyses revealed the higher levels of TH expression (20.3%) and dopamine secretion (265.5 $\pm$ 62.8 pmol/mg) in bFGF and TGF-sequentially treated hES cells than those in $\alpha$ RA or BDNF treated hES cells. Conclusion: These results indicate that the generation of dopamine secretory neurons from in vitro differentiated hES cells can be improved by TGF-$\alpha$ addition in the bFGF induction protocol.

• Journal of Korean Neurosurgical Society
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• v.30 no.4
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• pp.417-424
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• 2001

Objective : Adrenal medullary chromaffin cells are known to release analgesic substances such as opioides and catecholamines. Transplantation of them is a novel method that challenges current approaches in treating chronic pain. The transplantation of xenogeneic chromaffin cells into the central nervous system(CNS) supply antinociception in animals. In this study, we investigated the analgesic effects of rat adrenal medullary chromaffin cells transplanted into the CNS of the mouse. To study the antinociceptive efficacy of transplanted chromaffin cells, the survival of rat adrenal medullary chromaffin cells transplanted into the CNS of mouse was determined. Methods : The adrenal medullary chromaffin cells isolated from rat were transplanted into the striatum of mouse. These cells were confirmed of the release of Met-enkephalin and Leu-enkephalin by HPLC, and immunoblots for tyrosine hydroxylase(TH). Two weeks after transplantation, we performed immunohistochemistry for TH to determine the survival of implanted cells and assessed pain sensitivity at the same time. Results : The isolated rat adrenal medullary chromaffin cells were positive for anti-TH antibody and released Met-enkephalin and Leu-enkephalin more than rat endothelial cells. Transplanted rat chromaffin cells were stained with anti-TH antibody in striatum of mouse after 2 weeks. Pain sensitivity was reduced on the chromaffin cell-transplanted mouse compared to endothelial cell-transplanted mouse by the hot plate test. Conclusion : These results suggest that the rat chromaffin cells were suitably transplanted into the CNS of mouse. This approach could be used as a therapy for reducing of chronic pain induced by cancer or neuronal injury.