• 대한한방내과학회지
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• 제22권2호
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• pp.135-144
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• 2001

Objectives : The purpose of this investigation is to evaluate the effects of Woohwangcheongsim-won on neuronal death of hypoxic E18 cortical neuroblast. Methods : To evaluate the effect of Woohwangcheongsim-won on neuronal death caused by hypoxia, the survival rate of E18 cortical neuroblast was measured with MTT assay and the changes of several synaptic proteins and enzymes were investigated with the immunoblot assays. Results : The E18 cortical neuroblasts were added 50, 100, 500, 1,000, and $5,000{\mu}g/ml$ Woohwangcheongsim-won. They showed neurotoxicity, when the concentration of Woohwangcheongsim-won was above $1,000{\mu}g/ml$. The E18 cortical neuroblasts, which were added 50, 100, and $500{\mu}g/ml$ Woohwangcheongsim-won, were exposed 98% $N_2/5%\;CO_2$ for 3 hours to induce hypoxia, 3 days later, the survival rate of $50{\mu}g/ml$ Woohwangcheongsim-won was 141.5% when compared to the control group. On the immuneblot assays, the expressions of ${\alpha}$CaMKII, NR2A, NR28, PDE2, PSD-95, and eEF-$1{\alpha}$ were increased in normoxia, but those of NR2A, NR2B were decreased in hypoxia when compared to the control group. Conclusions : The data shows that the effects of Woohwangcheongsim-won on neuronal death of hypoxic E18 cortical neuroblast is a significant result.

• 대한한의학회지
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• 제22권1호
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• pp.63-77
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• 2001

Objectives : In acute stage of CVA, many patients experience Bugibultong as a symptom. Daesiho-tang is one of the most useful prescriptions for constipated stroke patients. This study investigated the effects of Daesiho-tang extracts on hypoxic E18 cortical neuroblast. Methods : To evaluate the effect of Daesiho-tang extracts on enuronal death caused by hypocia the neuronal viability and protein expressions of NMDA receptor subunits, ${\alpha}CaMK$ II PDE2 PSD95 densin-180, $eEF-l{\alpha}$ and GFAP were investigated. Results : We guessed that Daesiho-tang extracts worked to protect against cell damage through effecting on the protein expression of PSD(post synatpic density), especially increase of ${\alpha}CaMK$ II, $eEF-l{\alpha}$ PDE2, PSD95 and densin-180 Conclusions : According to the above results, it is supposed that Daesiho-tang is clinically applicable to stroke patients in the acute stage.

• 대한한방내과학회지
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• 제24권1호
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• pp.104-112
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• 2003

Objectives : The purpose of this study is to evaluate the effects of Sohaphyang-won and is to study the mechanism for neuronal death protection in hypoxia with Embryonic day 20(E20) cortical cells of a guinea pig(Sprague Dawley). Methods : E20 cortical cells, used in this investigation were dissociated in Neurobasal media and grown for 14 days in vitro (DIV). On 14 DIV, Sohaphyang-won was added to the culture media for 72 hours. On 17 DIV, cells were given a hypoxic shock and further incubated in normoxia for another three days. On 20 DIV, Sohaphyang-won's effects for neuronal death protection were evaluated by LDH assay and the mechanism was studied by Bcl-2, Bak, Bax, caspase family. Results : This study indicates that Sohaphyang-won's effects for neuronal death protection in hypoxia is confirmed by LDH assay by the method of Embryonic day 20(E20) cortical neuroblast. Conclusions : Sohaphyang-won's mechanism for neuronal death protection in hypoxia restrains inflow of cytochrome C into cellularity caused by Bcl-2 increase and reduces the caspase cascade initiator caspase-10 and the effector caspase-3.

• 대한한방내과학회지
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• 제24권2호
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• pp.348-357
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• 2003

Objectives : The purpose of this investigation is to evaluate the effects of Aucklandiae Radix Moschus(木香 麝香)and to study the mechanism for neuronal death protection in hypoxia with Embryonic day 20 (E20) cortical cells of a rat (Sprague Dawley). Methods : E20 cortical cells used in this investigation were dissociated in Neurobasal media and grown for 14 days in vitro (DIV). On 14 DIV, Aucklandiae Radix Moschus(木香 麝香) was added to the culture media for 72 hrs. On 17 DIV, cells were given a hypoxic shock and further incubated in normoxia for another three days. On 20 DIV, Moschus(麝香)'s effects for neuronal death protection were evaluated by LDH assay and the mechanisms were studied by Bcl-2, Bak, Bax, caspase family. Results : This study indicate that Aucklandiae Radix(木香)'s effects for neuronal death protection in normoxia and Scutellariae Radix(麝香)'s effects for neuronal death protection in hypoxia were confirmed by LDH assay in culture method of Embryonic day 20(E20) cortical neuroblast. Moschus(麝香)'s mechanism for neuronal death protection in hypoxia is to increase the anti-apoptosis protein Bcl-2. Conclusions : It may be reasonable to propose that Moschus(麝香) protects delayed neuronal death in hypoxia by increasing Bcl-2, thereby reducing mitochondrial permeability transition(PT) pores, the cytochrome c channels.