• 한국독성학회:학술대회논문집
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• 한국독성학회 2002년도 Molecular and Cellular Response to Toxic Substances
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• pp.14-40
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• 2002

15-Deoxy- Δ$\^$12,14/-prostaglandin J$_2$ (15-deoxy-PGJ$_2$), a naturally occurring ligand activates the peroxisome proliferator-activated receptor-${\gamma}$ (PPAR-${\gamma}$). Activation of PPAR-y has been found to induce cell differentiation such as adipose cell and macrophage. Here it was investigated whether 15-deoxy-PGJ$_2$ has neuronal cell differentiation and possible underlying molecular mechanisms. Dopaminergic differentiating PC 12 cells treated with 15-deoxy-PGJ$_2$ (0.2 to 1.6 ${\mu}$M) alone showed measurable neurite extension and expression of neurofilament, markers of cell differentiation. However much greater extent of neurite extension and expression of neurofilament was observed in the presence of NGF (50 ng/$m\ell$). In parallel with its increasing effect on the neurite extension and expression of neurofilament, 15-deoxy-PGJ$_2$ enhanced NGF-induced p38 MAP kinase expression and its phosphorylation in addition to the activation of transcription factor AP-1 in a dose dependent manner. Moreover, pretreatment of SD 203580, a specific inhibitor of p38 MAP kinase inhibited the promoting effect of 15-deoxy-PGJ$_2$ (0.8 ${\mu}$M) on NGF-induced neurite extension. This inhibition correlated well with the ability of SB203580 to inhibit the enhancing effect of 15-deoxy-PGJ$_2$ on the expression of p38 MAP kinase and activation of AP-1. The promoting ability of 15-deoxy-PGJ$_2$ did not occur through PPAR-${\gamma}$, as synthetic PPAR-${\gamma}$ agonist and antagonist did not change the neurite promoting effect of 15-deoxy-PGJ$_2$. In addition, contrast to other cells (embryonic midbrain and SK-N-MC cells), PPAR-${\gamma}$ was not expressed in PC-12 cells. Other structure related prostaglandins, PGD$_2$ and PGE$_2$ acting via a cell surface G-protein-coupled receptor (GPCR) did not increase basal or NGF-induced neurite extension. Moreover, GPCR (EP and DP receptor) antagonists did not alter the promoting effect of 15-deoxy-PGJ$_2$ on neurite extension and activation of p38 MAP kinase, suggesting that the promoting effect of 15-deoxy-PGJ$_2$ may not be mediated GPCR. These data demonstrate that activation of p38 MAP kinase in conjunction with AP-1 signal pathway may be important in the promoting activity of 15-deoxy-PGJ$_2$ on the differentiation of PC12 cells.

• 동의신경정신과학회지
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• 제11권1호
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• pp.1-18
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• 2000

To study the effects of Ramulus et Uncus Uncariae (REUU) on oxygen free radical-mediated damage by hydrogen peroxide $(H_{2}O_{2})$ on cultured spinal sensory neurons, in vitro assays such as MTT assay, NR assay, neurofilament enzymeimmuno assay (EIA), sulforhodamine B (SRB) assay, assay for lactate dehydrogenase (LDH) activity and assay for lipid peroxidation were used in cultured spinal dorsal root ganglion neurons derived from mice, Spinal dorsal root ganglion neurons were cultured in media containing various concentrations of $H_{2}O_{2}$ for 5 hours, after which the neurotoxic effect of $H_{2}O_{2}$ was measured by in vitro assay. The protective effect of the herb extract, Ramulus et Uncus Uncariae (REUU) against H2O2-induced neurotoxicity was also examined. The results are as follows. 1. In NR assay and MTT assay, $H_{2}O_{2}$ significantly decreased the cell viability of cultured mouse spinal dorsal root ganglion neurons according to exposure concentration in these cultures. An additional time course study was done on these cultures. 2. Cultured spinal dorsal root ganglion neurons which were exposed to various concentrations of $H_{2}O_{2}$ showed a quantitative decrease of neuronal cells by EIA and of total protein by sulforhodamine B (SRB) assay, while they showed an increase of both lipid peroxidation and LDH activity. 3. The effect of Ramulus et Uncus Uncariae (REUU) on $H_{2}O_{2}$ induced neurotoxicity showed a quantitative increase in both neurofilament and total protein, but showed a decrease of lipid peroxidation and LDH activity. These results suggest that $H_{2}O_{2}$ has a neurotoxic effect on cultured spinal dorsal root ganglion neurons from mice and that the herb extract, Ramulus et Uncus Uncariae (REUU), was very effective in protecting $H_{2}O_{2}$ induced neurotoxicity by decreasing lipid peroxidation and LDH activity.

• 대한한방내과학회지
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• 제24권1호
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• pp.11-20
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• 2003

Objective : Experimental studies have been done to elucidate the effect of kangwhalyupung-tang(KWYPT) on neuronal cell damage induced by brain ischemia. Method : The cytotoxic effect of ischemia was measured in the MTS assay cultures. MTS assay, INT assay, neurofilament(NF) enzymeimmuno assay(EIA). And the KWYPT on ischemia-induced neurotoxicity were examined by in vitro assays. Results : 1. The KWYPT protected effectively neuronal cell-death resulted from brain ischemia induced by the treatment of $95%N_2/5%CO_2$ for 10 min in those dependent fashion. 2. The KWYPT effectively increased the amount of NF resulting from brain ischemia, induced by the treatment of $95%N_2/5%CO_2$ for 15 min in those dependent fashions. Conclusions : KWYPT protects the brain ischemia-induced neurotoxicity through the increase of cell viability and of neurofilament in dose-dependent manner.

• 동의생리병리학회지
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• 제24권4호
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• pp.681-687
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• 2010

Sibjeondaebo-tang (SJDBT) is an oriental medicinal prescription for the treatments of diverse symptoms including neurological disorders. In order to investigate its potential role for neural regulation following nerve injury, neurite outgrowth of dorsal root ganglion (DRG) neurons in culture was investigated. In DRG neurons which were preconditioned by sciatic nerve injury, neurite outgrowth was enhanced by SJDBT treatment. When preconditioned DRG neurons were co-cultured with astrocytes prepared from injured spinal cord tissue, neurite outgrowth was similarly facilitated by SJDBT. Astrocytes in co-culture showed more intense signals of vimentin protein by SJDBT compared to saline control. Sukjihwang (SJH), a conventional herbal component of SJDBT prescription, did not induce any significant changes in neurite extension of DRG neurons compared to control cells. These data suggest that SJDBT may be the therapeutic agent for nervous system disorders related to nerve damage.

• 한국정보통신학회:학술대회논문집
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• 한국정보통신학회 2014년도 추계학술대회
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• pp.989-992
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• 2014

쥐에서 슈반세포와 뉴런세포를 이용한 수초화가 수행되었다. 슈반세포와 뉴런 세포는 쥐의 배아(임신 16일)의 척수신경절로 부터 각각 분리되어 배양되었다. 배아의 척수신경절이 배양되었고 항 유사분열제가 첨가되었다. 분리 정제된 배아의 슈반세포가 배양되었고 이것은 분리 정제된 배아의 척수신경절 세포에 첨가되었다. 실험실상에서 분리 정제된 수초화 군을 완성할 수 있었다. 뉴로필라멘트 단백질의 항체를 이용하여 수초화의 형성되었음을 확인하였다.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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• pp.169.2-170
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• 2003

Erythropietin (EPO), a hematopoietic factor is also required for normal brain development, and its receptor is localized in brain. Therefore, it is possible that EPO could act as a neurotropic factor inducing differentiation of neurons. The present study, we therefore investigated whether EPO can increase differentiation of undifferentiated cortical neuron isolated from postneonatal (Day 1) rat brains and PC12 cell, undifferentiated dopaminagic cell line. EPO dose (1-100 U/ml) dependently increased cell differentiation and expression of differentiation marker gene (neurofilament and tyrosine hydroxylase) in both cells. (omitted)

• 대한소아치과학회지
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• 제21권2호
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• pp.439-455
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• 1994

생리적 치근 흡수가 뒤따르는 유치 치주인대의 신경분포 및 변화를 평가하기 위하여 생후 1, 2. 3. 4. 5개월된 개의 유치 치주인대와 9개월된 영구치 치주인대를 대상으로 neurofilament protein(NFP) 을 이용한 면역조직화학적 방법으로 관찰하여 다음과 같은 결과를 얻었다. ${\bullet}$ 유견 유치의 치주인대 신경분포는 치근단 1/3 부위에서 가장 밀집되어 있었으며 치근 중앙부로 향할수록 현저하게 감소되었으며 전체적으로 그 분포밀도 및 분지 정도는 영구치와 비교시 약한 양상을 나타냈다. ${\bullet}$ 구치부에 비해 전치부에서 신경분포가 보다 발달된 양상을 보였으며 구치 치근이개부 치주인대에서는 신경섬유가 관찰되지 않았다. ${\bullet}$ 유치근이 흡수됨에 따라 신경분포 정도도 감소되었으며 탈락시기의 치근 주위에서는 신경섬유를 관찰할 수 없었다. ${\bullet}$ 치은점막상피의 신경지배는 치아가 발육함에 따라 감소되는 양상을 나타내었으며 설측점막이 순측점막보다 더 발달된 신경분포 양상을 관찰할 수 있었다. ${\bullet}$ 유치 치주인대 신경말단 형태는 수지상의 신경말단 형태를 나타내고 있었으며 전형적인 Ruffini 신경말단은 거의 관찰할 수 없었다. 이상의 결과를 미루어 신경섬유의 분포는 기능과 밀접한 관계가 있는 것으로 생각되고 더우기 유치의 생리적 치근흡수에 따른 치주인대에 신경섬유의 분포 및 변화는 유치의 성장발육에 따른 하나의 변화라고 생각된다.

• Applied Microscopy
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• 제35권4호
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• pp.55-63
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• 2005

쥐의 뇌로부터 미세혈관에서 분리하고 배양한 내피세포의 특성을 현미경관찰, 면역염색과 전기저항을 측정해 관찰하였다. 미세혈관 내피세포는 배양 후 $5{\sim}6$일 경에 단층을 형성하였으며 특징적으로 소용돌이치는 모양을 나타냈다. 내피세포 단층의 전기저항은 배양 후 5일 경까지 따라 증가하였고 이후로는 감소하였다. 면역형광염색에서 anti-GFAP, anti-GalC, anti-neurofilament 160/200 kD antibody에 대한 면역반응을 거의 찾아볼 수 없었어 별아교세포, 희소돌기아교세포 및 신경세포에 의한 우려할 만한 오염은 배제할 수 있었다. vWF 항원에 대한 면역반응은 내피세포의 세포질에 Weibel-Palade 과립이 전반적으로 퍼져 있었다. 치밀이음부를 구성하는 occludin, ZO-1, ZO-2에 대한 면역반응은 내피세포의 접촉부위에서 매우 특징적으로 나타나고 있었다. 요약하면 쥐의 뇌 미세혈관 내피세포를 분리, 배양하여 형태학적, 조직면역 화학적 방법과 전기저항을 측정하여 내피세포의 특성을 확인하였다. 이 생체외 혈액뇌장벽 모델은 앞으로 진행될 액뇌장벽의 특징을 규명하고자 하는 시험관내 실험에 유용하게 이용될 수 있을 것이다.

• 대한한의학회지
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• 제33권3호
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• pp.133-146
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• 2012

Background: Most antitumor agents have the side effect of chemotherapy-induced peripheral neuropathy (CIPN). Cancer patients who take antitumor agents suffer from CIPN, but there is no known treatment for it. Unlike the central nerve system, the peripheral nerve can self-repair, and the Schwann cell takes this mechanism. Objectives: In this study, we researched the effect of YideungJetong-Tang (YJT) extract on taxol-induced sciatic nerve damage, through in vitro and in vivo experiments. Also, we studied the effect of YJT extract on neurite recovery and anti-inflammatory effect after compression injury of sciatic nerve in vivo. Methods: Vehicle, taxol and taxol+YJT were respectively applied on sciatic nerve cells of rat in vitro, then the cells were cultured. The sciatic nerve cells and Schwann cells were then observed using Neurofilament 200, Hoechst, ${\beta}$ -tubulin, S-$100{\beta}$, caspase-3 and phospho-Erk1/2. CIPN was induced by taxol into the sciatic nerve of rat in vivo, then YJT extract was taken orally. The axons, Schwann cells and neurites of the DRG sensory nerve were then observed using Neurofilament 200, ${\beta}$-tubulin, Hoechst, S-$100{\beta}$, phospho-Erk1/2 and caspase-3. YJT was taken orally after sciatic nerve compression injury, and the changes in axon of the sciatic nerve, Schwann cells and TNF-${\alpha}$ concentration were observed. Results: The taxol and YJT treated group showed significant effects on Schwann cell recovery, neurite growth and recovery. In vivo, YJT compared with control group showed Schwann cell structural improvement and axons recovering effect after taxol-induced Schwann cell damage. After sciatic nerve compression injury, recovery of distal axon, changes of Schwann cell distribution, and anti-inflammatory response were observed in the YJT. Conclusions: Through this study, we found that after taxol-induced neurite damage of sciatic nerve in vivo and in vitro, YJT had significant effects on sciatic nerve growth and Schwann cell structural improvement. In vivo, YJT improved recovery of distal axons and Schwann cells and had an anti-inflammatory effect.

• 대한소아치과학회지
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• 제23권2호
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• pp.291-305
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• 1996

유치 및 초기 영구치의 치수신경분포 및 변화를 평가하기 위하여 교정치료를 위해 발치한 치근흡수가 비교적 적은 유치, 정상 탈락기의 유치, 맹출중인 교합하지 않은 제 1소구치, 맹출이 완료되어 교합하고 있는 제 1소구치를 대상으로 neurofilament protein(NFP) 을 이용 한 변역조직화학적 방법으로 관찰하여 다음과 같은 결과를 얻었다. 치근흡수가 비교적 적은 유치에서도 영구치와 유사한 신경총의 존재와 신경분포 형태가 관찰되었으나, 영구치에 비해 그 신경분포의 밀도가 낮았으며 분지형성 또한 미약하게 나타났고, 전상아질로 진입하는 섬유의 수가 적었다. 정상탈락기의 유치에서는 신경의 변성으로 소포(vesicle), 단절(fragmentation)등이 관찰되었으며, 그 신경분포가 현저히 감소되어 나타났고, 총상의 형태도 소실되었다. 교합하지 않은 제 1소구치에서 치수 내의 신경은 말단가지의 숫자가 적고, 치수 조상아 세포 경계에 도달하였으며, 비교적 가는 신경 섬유가 많았다. 교합하고 있는 제 1소구치에서 치수 내의 신경은 말단이 조상아층으로 진출하고, 어떤 신경섬유는 전상아질층까지 진입하였으며, 비교적 굵은 신경섬유의 분포가 많아졌다.