• 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.)

• 한국감성과학회:학술대회논문집
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• 한국감성과학회 2008년도 추계학술대회
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• pp.147-150
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• 2008

Vitamin C ascorbic acid (AA) and dehydroascorbic acid (DHA) as an antioxidant have been shown to have protective effects in experimental neurological disorder models such as stroke, ischemia, and epileptic seizures. The present study was conducted to examine the protective effect of AA and DHA on Kainic acid (KA) neurotoxicity using organotypic hippocampal slice cultures (OHSC). After 12h KA treatment, significant delayed neuronal death was detected in CA3 region, but not in CA1. Intermediate dose of AA and DHA pretreatment significantly prevented cell death and inhibit ROS level, mitochondrial dysfunction and capase-3 activation in CA3 region. In the case of low or high dose, however, AA or DHA pretreatment were not effective. These data suggest that both AA and DHA pretreatment have neuroprotective effects on KA-induced neuronal injury depending on the concentration, by means of inhibition of ROS generation, mitochondrial dysfunction, and caspase-dependent apoptotic pathway.

• 대한한방내과학회지
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• 제25권4호
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• pp.227-241
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• 2004

Objective : This study was performed to investigate neuroprotective effects of Bupleuri Radix against oxidative and ischemic damages. Method : To observe the neuroprotective effects against ischemic damage, ischemic insult was induced by oxygen/glucose deprivation (OGD) on organotypic hippocampal slice cultures (OHSC) from 1 week-old Sprague-Dawley rats. Propidium iodide (PI) fluorescence-stained neuronal dead-cell areas, area percentages and TUNEL-positive apoptotic cells in CA1 and dentate gyrus, and LDH levels in culture media of the OHSC were measured following Bupleuri Radix extract treatment. Result : The following results were obtained: (1) The $5\;{\mu}g/ml$ of Bupleuri Radix treatment demonstrated a significant decrease in PI fluorescence-stained neuronal dead-cell areas and area percentage in CA1 region of the OHSC from 18 hrs to 48 hrs following the OGD. The $50\;{\mu}g/ml$ of Bupleuri Radix treatment was also significant from 6 hrs to 48 hrs following the OGD and was more effective. (2) The 5 and $50\;{\mu}g/ml$ of Bupleuri Radix treatment demonstrated a significant decrease in PI fluorescence-stained neuronal dead-cell areas and area percentage in DG region of the OHSC from 6 hrs to 48 hrs following the OGD. The $50\;{\mu}g/ml$ treatment was more effective than the $5\;{\mu}g/ml$ treatment. (3) Bupleuri Radix treatment demonstrated a significant decrease in TUNEL-positive apoptotic cells in CA1 region (with 5 and $50\;{\mu}g/ml$) and in DG region (with $50\;{\mu}g/ml$) of the OHSC damaged by the OGD. (4) Bupleuri Radix treatment demonstrated a significant decrease in LDH concentrations in culture media of the OHSC damaged by the OGD. Conclusion : These results suggest that Bupleuri Radix has neuroprotective and control effects on inflammatory and immune responses where there has been ischemic damage to the central nervous system.