• 대한한의학회지
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• 제23권3호
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• pp.164-173
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• 2002

목적：반응성산소기들은 장관에서 여러 종류의 질병의 발생과 관련을 가지고 있는 것으로 알려져 있어, 이들에 의한 세포손상을 방지하는 약물의 개발은 시급한 실정이다. 본 연구에서는 항산화작용을 가진 약재로 보고 된 단삼추출액이 장관상피세포에서 $H_2O_2$에 의한 세포손상을 방지할 수 있는 지를 조사하고자 하였다. 방법：장관상피세포로는 사람의 소장상피세포에서 유래한 배양세포주인 Caco-2세포를 이용하였고, 세포손상 정도는 trypan blue exclusion assay를 통해 평가하였고, 지질의 과산화는 그 산물인 malondialdehyde의 량을 측정하여 산정하였다. 결과： $H_2O_2$는 처리 시간 및 농도에 비례하여 세포손상을 유발하였으며, 이러한 효과는 단삼추출액에 의해 농도의존적으로 방지되었다. $H_2O_2$에 의한 세포소상은 $H_2O_2$제거제인 catalase와 철착염제인 deferoxamine에 의해 방지되었으나 항산화제인 N,N-diphenyl-p-phenylenamine(DPPD)에 의해 영향을 받지 않았다. $H_2O_2$는 지질의 과산화를 증가시켰으며, 이러한 효과는 단삼추출액과 DPPD에 의해 억제되었다. 단삼추출액은 $H_2O_2$에 의한 세포내 ATP 고갈을 방지하였다. $H_2O_2$는 DNA 손상을 일으켰으며, 이러한 효과는 단삼추출액, catalase 및 deferoxamine에 의해 방지되었으나, DPPD에 의해서는 변화되지 않았다. 결론 : 이상의 결과를 종합하면 단삼추출액은 장관상피세포에서 $H_2O_2$에 의한 세포손상을 방지하며, 이러한 효과는 항산화작용이 아닌 다른 작용기전에 기인할 것으로 생각된다. 또한 본 연구의 결과는 $H_2O_2$가 장관상피세포에서 지질의 과산화를 유발하여 세포손상을 일으키지 않음을 가리킨다.

• 대한한의학방제학회지
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• 제11권2호
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• pp.159-171
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• 2003

This study was done to investigate effects of SR, CF and AR to angiogenesis of cerebral tissues, protecting damage of cerebral neurons and activating them in cerebral contusion-induced rats. I observed these conclusions as follows ; 1. Observation of VEGF-immunoreactive cells : Groups of administered AR were not meaningful in increasing VEGF-immunoreactive cells for 3 days and 7 days, groups of administered SR meaningfully increased them to control groups in all groups, and groups of administered CF meaningfully increased them to control groups in all groups, too, interestingly, increased double to control group for 7 days. 2. Observation on cerebral neurons by Cresyl violet stain : Dendrites and axons of groups of administered SR, AR for 3 days were clearly observed to control group. Cerebral neurons of groups of administered CF for 3days and 7 days were increased a little, but were not meaningful. In conclusion, AR will be careful of being used in cerebral contusion. CF and SR were effective to activating cerebral hemokinesis by inducing angiogenesis in trauma of tissue, but weakly to protecting trauma of cerebral neurons and activating them. I think more studies will be done in these facts.

• 대한한방내과학회지
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• 제25권3호
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• pp.461-472
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• 2004

Objectives : For the screening of neuroprotective effects of medicinal herbs, the complex system of animal models suffer some disadvantages in controlling critical parameters such as blood pressure and body temperature. Additionally, application of drugs to the appropriate brain area sometimes is difficult, due to poor permeability though the blood brain barrier, and so potential protective effects might be masked. Methods : Organotypic hippocampal slice culture (OHSC) method has the advantages of being relatively easy to prepare and of maintaining the general structure, including tissue integrity and the connections between cells. Drugs can easily be applied and neuronal damage can easily be quantified by using tissues and culture media. This study demonstrates neuroprotective effects of Puerariae radix (葛根, PR), Salviae miltiorrhizae radix (丹蔘, SR), Rhei rhizoma (大黃, RR), and Bupleuri radix (柴胡, BR). These were screenedand compared to MK-801, antagonist of NMDA receptors, by using OHSC of 1 week-old Sprague-Dawley rats. Oxygen/glucose deprivation (OGD) were conducted in an anaerobic chamber $(85%\;N_2,\;10%\;CO_2\;and\;5%\;H_2)$ in a deoxygenated glucose-free medium for 60 minutes. Water extracts of each herbs were treated to culture media with $5\;{\mu}g/ml$ for 48 hours. Results : Neuronal cell death in the cultures was monitored by densitometric measurements of the cellular uptake of propidium iodide (PI). PI fluorescence images were obtained at 48 hours after the OGD and medicinal herb treatment. Also TUNEL-positive cells in the CAI and DG regions and LDH concentrations in culture media were measured at 48 hours after the OGD. According to measured data, MK-801, PR, SR and BR demonstrated significant neuroprotective effect against excessive neuronal cell death and apoptosis induced by the OGD insult. Especially, PR revealed similar neuroprotective effect to MK-801 and RR demonstrated weak neuroprotective effect. Conclusions : These results suggest that OHSC can be a suitable method for screening of neuroprotective effects of medicinal herbs. (This work was supported by the research program of Dongguk University and Grant 01-PJ9-PG1-01CO03-0003 from Ministry of Health & Welfare.)