• Applied Microscopy
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• v.39 no.1
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• pp.1-7
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• 2009

Experimental induction of polycystic ovary (PCO) resembling some aspects of human PCO syndrome was produced using the long-acting compound estradiol valerate (EV). Our previous study on the role of Korean red ginseng total saponins (GTS) in a steroid-induced PCO rat model demonstrated that electro-acupuncture modulates nerve growth factor (NGF) concentration in the ovaries. In fact, the involvement of a neurogenic component in the pathology of PCO-related ovarian dysfunction is preceded by an increase in sympathetic outflow to the ovaries. In the present study, we tested the hypothesis that therapeutic GTS administration modulates sympathetic nerve activity in rats with PCO. This was done by analyzing NGF protein and NGF mRNA expression involved in the pathophysiological process underlying steroid-induced PCO. EV injection resulted in significantly higher ovarian NGF mRNA expression in PCO rats compared to control rats, and PCO ovaries were counteracted by GTS administration with significantly lower expression of NGF mRNA compared to EV treated ovaries. However, NGF protein was unaffected in both EV and GTS treated ovaries compared to control rats. These results indicate that EV modulates the neurotrophic state of the ovaries, which may be a component of the pathological process by which EV induces cyst formation and anovulation in rodents.

• Applied Microscopy
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• v.41 no.2
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• pp.109-116
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• 2011

Polycystic ovary syndrome (PCOS) is hormonal imbalance condition as the endocrine and metabolic disorder that induces the infertility and various complications in reproductive age women. Estradiol valerate (EV) is used hormone replacement therapy in menopausal women and is reported that excessive administration of EV induces the PCOS. Nerve growth factor (NGF) is the factor to regulate the survival and maturation of developing neuronal cell and is also synthesized in ovary. And NGF is overexpressed in EV-induced polycystic ovary (PCO) as previously reported. Therefore, this study examined the possibility of NGF as can be used the biological marker in diagnosis of PCOS, the hormonal imbalance condition, using PCO and CHO (chinese hamster ovarian) cell lines. The concentration of EV treatment is optimized a 1 mg as not influence on the proliferation of CHO cell but 2 mg and 3 mg of EV treatment have the inhibition effect at initial stage. The morphological change was not observed in CHO cell after dose dependent manner treatment of EV. Expression of NGF mRNA and protein is significantly increased at 30 min after EV treatment in CHO cells compared to that of control. And NGF protein expression is strongly increased in PCO tissue, which observed many follicular cysts compared to normal ovary tissue. Taken together, overexpression of NGF may be act as a molecule to induce an abnormal development of follicle, suggesting that NGF can be used as a biological marker in diagnosis of PCOS.

• Journal of Magnetics
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• v.20 no.4
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• pp.377-380
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• 2015

Muscle contusion has a negative effect on muscle function. Although several studies showed that pain control and muscle recovery is facilitated by pulsed electromagnetic fields (PEMF), there has not been much research regarding the specific effects of PEMF on them. The aim of the present study is to investigate effects of PEMF on pain and muscle recovery following extensor digitorum longus (EDL) contusion injury through measuring the expression of the c-fos proto-oncogene and nerve growth factor (NGF). Significantly reduced c-fos expression in the spinal cord was shown in PEMF groups compared with control (CON) groups. There was no significant difference between PEMF1 and CON1, but significantly increased NGF expression was shown in PEMF3 and PEMF5 compared with in CON groups, where the numbers in the group names are the days from contusion. In conclusion, PEMF could be used to not only reduce pain in muscle injuries by down-regulating c-fos expression in the spinal cord, but it could also influence muscle healing through increasing NGF expression in the injured muscle.

• BMB Reports
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• v.40 no.6
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• pp.888-894
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• 2007

Src homology (SH) domains of phospholipase C-$\gamma1$ (PLC-$\gamma1$) impair NGF-mediated PC12 cells differentiation. However, whether the enzymatic activity is also implicated in this process remains elusive. Here, we report that the enzymatic activity of phospholipase C-$\gamma1$ (PLC-$\gamma1$) is at least partially involved to the blockage of neuronal differentiation via an abrogation of MAPK activation, as well as sustained Akt activation. By contrast, Overexpression of WT-PLC-$\gamma1$ exhibited sustained NGF-induced MAPK activation, and triggered transient Akt activation resulting in profound inhibition of neurite outgrowth. However, lipase-inactive mutant (LIM) PLC-$\gamma1$ cells fail to suppress neurite outgrowth, although it contains intact SH domains, specifically enhancing the expression of cyclin D1 and p21 proteins, which regulate the function of retinoblastoma Rb protein. These observations show that the lipase inactive mutant of PLC-$\gamma1$ does not alter NGF-induced neuronal differentiation via enzymatic inability and the modulation of cell cycle regulatory proteins independent on SH3 domain.

• Korean Journal of Medicinal Crop Science
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• v.18 no.5
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• pp.291-297
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• 2010

Liriope platyphylla has been though as an useful medical plant to improve the cough, sputum, neurodegenerative disorders, obesity, and diabetes in Korea and China from old times. In order to investigate the effects of Liriope platyphylla on expression and secretion of nerve growth factor (NGF), the mRNA expression and protein secretion were detected in the neuronal cell (B35) and neuroglial cell (C6) cultured with three differences concentration (5%, 10%, 15%) of Liriope platyphylla. In MTT assay and FACS anslysis, the some death of some B35 and C6 cells were observed in 15% extract-treated group, while other groups did not induce the death. Also, the mRNA expression of NGF were significantly increased in 5% and 10% extracts treated-group. Furthermore, the NGF protein concentration in supernatant collected from cultured cells showed the very similar pattern with mRNA expression. In order to verify the activity of secreted NGF, the culture supernatant collected from B35 and C6 cells cultured with Liriope platyphylla extracts for 24 hrs were treated into undifferentiated PC12 cells, and the differentiation level of PC12 cell were also observed with microscopes. The differentiation level of PC12 cell were significantly increased depend on the dose of extract. Therefore, these results suggested that the water extracts of Liriope platyphylla may contribute the regulation of NGF expression and secretion in the neuronal cell and be considered as an excellent candidate for a neurodegenerative disease-therapeutic drug.

• The Journal of Korean Obstetrics and Gynecology
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• v.24 no.3
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• pp.48-72
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• 2011

Objectives: This study was designed to investigate the effects of Yongdamsagantang on the polycystic ovary(PCO) induced by estradiol valerate(EV) in rats. Methods: After administrating Yongdamsagan-tang to PCO induced rats, we measured the weight of body, ovaries, adrenal glands, and uterus of rats. The observation through naked eye and histopathological observation of ovaries were evaluated. Also, the number of follicle and corpora lutea and content of androstenedione(ADD) and total estrogen were evaluated. The expressions of nerve growth factor(NGF) and corticotropin releasing factor(CRF) were analyzed by immunohistochemistry. The breeding rate and number of implantation with normal male rats were evaluated. Results: - The weight(mg) of ovaries in YST treated group($73.8{\pm}7.6$) was significantly increased(p<0.001) compared with control group($54.3{\pm}4.5$). - The number of mature follicles in YST treated group($7.3{\pm}2.4$) was significantly increased(p<0.01) compared with control group($3.5{\pm}1.2$). - The number of atretic follicles in YST treated group($9.0{\pm}1.5$) was significantly decreased(p<0.01) compared with control group($13.4{\pm}3.8$). - The number of cystic follicles in YST treated group($3.1{\pm}1.1$) was significantly decreased(p<0.01) compared with control group($6.0{\pm}2.0$). - The number of corpora lutea in YST treated group($3.8{\pm}2.1$) was significantly increased(p<0.001) compared with control group($0.3{\pm}0.7$). - The expression of NGF-immunoreactive cells in the ovarian granulosa cells in YST treated group was lesser observed than control group. - The expression of NGF-immunoreactive cells in the adrenal cortex in YST treated group was lesser observed than control group. - The breeding rate in YST treated group(100 %) was significantly increased (p<0.05) compared with control group(50 %). - The number of implantation in YST treated group($6.4{\pm}4.7$) was significantly increased(p<0.05) compared with control group($1.4{\pm}2.6$). Conclusions: We concluded that Yongdamsagan-tang activates the maturation of follicles, normal ovulation, breeding rate and number of implantation. Therefore, this may be effective for the treatment of anovulation, amenorrhea and sterility of PCOS patients.

• Journal of Life Science
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• v.21 no.8
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• pp.1134-1141
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• 2011

Transmembrane protein 21 (TMP21) is a member of the p24 cargo protein family and has been shown to modulate ${\alpha}$-secretase-mediated A${\beta}$ production which was specifically observed in the brains of subjects with Alzheimer's disease (AD). In order to investigate whether TMP21 could affect nerve growth factor (NGF) receptor signaling pathway, the alteration of NGF receptors and their downstream proteins were detected in TMP21 over-expressed cells. CMV/hTMP21 vector used in this study was successfully expressed into TMP21 proteins in B35 cells after lipofectamin transfection. Expressed TMP21 proteins induced the down-regulation of ${\gamma}$-secretase complex components including Presenlin-1 (PS-1), PS-2, Nicastrin (NST), Pen-2 and APH-1. Also, the expression level of NGF receptor $p75^{NTR}$ and RhoA were significantly higher in CMV/hTMP21 transfectants than vehicle transfectants, while their levels returned to vehicle levels after NGF treatment. However, the phosphorylation of NGF receptor TrkA was dramtically decreased in NGF No-treated CMV/hTMP21 transfectants compared with vehicle transfectants, and increased in NGF treated CMV/hTMP21 transfectants. In TrkA downstream signaling pathway, the phosphorylation level of ERK was also decreased in CMV/hTMP21 transfectants, while the phosphorylation of Akt was increased in the same transfectants. Furthermore, NGF treatment induced the increase of phosphorylation level of Akt and ERK in CMV/hTMP21 transfectants. Therefore, these results suggested that over-expression of TMP21may simultaneously induce the up-regulation of $p75^{NTR}$/RhoA expression and the down-regulation of TrkA/ERK phosphorylation through the inhibition of ${\gamma}$-secretase activity.

• The Korean Journal of Zoology
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• v.36 no.2
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• pp.245-263
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• 1993

신경성장인자 수용체(nerve growth factor receptor, HGFr)의 소재를 휜쥐 전뇌 기저부 핵들의 신경세포와 그 세포내 소기 관에서 연역세포화학적 방법으로 관찰하였다. NGFr에 면역반응을 보이는 신경세포들은 내측중격, 수직 및 수평대각선 브로카대, 거대세포 시삭전핵 그리고 Meynert 기저핵에는 다수 미상핵-피각과 복부담창구에는 소수 관찰 되었다 NGFr에 면역반응을 보이는 신경세포들은 형태학적으로 3가지 형 즉, 1) 난형(또는 원형). 2) 방추형, 3) 삼각형(또는 다각형)으로 구분되었다 내측중격은 주로 난형의 세포로 구성되었으며(91.2%), 수직 및 수평대각선 브로카대, 거대세포 시삭전핵 및 Meynert 기저 핵에는 난형의 세포가 높은 율로 구성되었으나, 방추형과 삼각형 세포들도 내측중격에서보다는 많았다 특히 복부담창구에는 다른 핵들에 비하여 방추형세포(25%)들이 높은 출현율을 보였다 일반적으로 이들 세포의 크기는 삼각형세포가 제일 컸으며, 방추형세포가 그 다음, 그리고 난형 세포가 제일 작았다 전자현미경적 관찰에서 0.05% triton X-100을 처리한 조직중 Meynert 기저핵을 관찰한 결과. Golgi체, multivesicular body 및 소포체들이 N6Fr에 면역반응을 보였으며. trion X-100을 처리하지 않은 조직에서는 단지 수평대각선 브로카대의 신경세포 원형질 막에서만 약한 면역반응을 보였다 위의 결과로 미루어 NGFr은 조연소포체에서 합성되어. Golgi체에서 농축되고, multivesicular body를 통하여 원형질막에 위치하게 되며, 원형질막에서 NGFr은 외래성의 NGF와 복합체를 형성한후, 궁극적으로는 Iysosome의 형태로 세포체 안으로 들어 가는 것으로 추정된다.

• 한국생물공학회:학술대회논문집
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• 2005.04a
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• pp.103-107
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• 2005

It has been known that the novel diterphenoids, hericenone and erinacine isolated from the fruiting body and cultured mycelia of Hericium erinaceus showed potent stimulating activity of nerve growth factor (NGF)-synthesis. To investigate the biological activities of extracts from fruiting body, cultured mycelium and cell-free broth of H. erinaceus, the activity experiments of antioxidation and AChE inhibition were carried out. Sephadex G-10 gel filtration followed by HPLC on a ${\mu}Bondapak$ $C_{18}$ column of EtOAc extract from cultured mycelium showed a biological activity.

• BMB Reports
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• v.31 no.5
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• pp.468-474
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• 1998

Membrane depolarization in PC12 cells induces calcium influx via an L-type voltage-sensitive calcium channel (L-VSCC) and increases intracellular free calcium, which leads to tyrosine phosphorylation of epidermal growth factor (EGF) receptor and the associated adaptor protein, She. This activated EGF receptor complex then can activate mitogen-activated protein (MAP) kinase, as in nerve growth factor (NGF) receptor activation. In the present study, we investigated the role of EGF receptor in the signaling pathway initiated by membrane depolarization of PC12 cells. Prolonged membrane depolarization induced phosphorylation of extracellular signal-regulated kinase (ERK) within 1 min in undifferentiated PC12 cells. Pretreatment of PC12 cells with the calcium chelator EGTA abolished depolarization-stimulated ERK phosphorylation, but NGF-induced phosphorylation of ERK was not affected. The chronic treatment of phorbol ester, which down-regulated the activity of protein kinase C (PKC), did not affect the phosphorylation of ERK upon depolarization. In the presence of an inhibitor of EGF receptor, neither depolarization nor calcium ionophore increased the level of ERK phosphorylation. These data imply that the EGF receptor is functionally necessary to activate ERK and neurite outgrowth in response to the prolonged depolarization in PC12 cells, and also that PKC is apparently not involved in this signaling pathway.