• 동의생리병리학회지
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• 제17권2호
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• pp.503-509
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• 2003

The purpose of this study is to examine the toxic effects caused by xanthine oxidase/hypoxanthine (XO/HX) and the effects of herbal extracts such as Mangeum-tang (萬金湯: MGT) and Gamimangeum-tang (加味萬金湯: GMGT) on the treatment of the toxic effects. The results of these experiments were XO/HX, an oxygen radical-generating system, decreased the survival rates of the cultured cells on XTT assay, the amount of DNA syntheses, and the amount of neurofilaments, and increased c-fos positive cells, MGT and GMGT have the efficacy of increasing the survival rates of the cultured cells by increasing the amount of neurofilaments and DNA synthesis and decreasing the c-fos positive cells damaged by XO/HX, From the above results, it is suggested that MGT and GMGT have marked efficacy as a treatment for the damages caused by the XO/HX-mediated oxidative stress. And MGT and GMGT are thought to have certain pharmacological effects.

• 동의생리병리학회지
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• 제17권4호
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• pp.1031-1036
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• 2003

To certify the protective effect of herbal medicine against oxygen free radical-induced myocardiotoxicity, cytotoxicity was measured using TBARS assay and Beating rate in the presence of Tongryeong-san(TRS) extracts or single constituents of this prescription. Myocardial toxicity was evaluated in neonatal rat myocardiocytes in cultures. In the present study, xanthine oxidase/hypoxanthine (XO/HX) resulted in a increase in lipid peroxidation and decreases in beating rate in cultured myocardial cells. In the effect of TRS extract, it showed the prevention from the XO/HX-induced cardiotoxicity by the increases of beating rate as well as the decrease of lipid peroxidation, In the protective effect of Faeces Trogopterori(FT), Pollen Typhae(PT), Caulis Akebiae(CA) and Radix Paeoniae Rubra(PRR), all the extracts were significantly effective in the protection of XO/HX-induced cardiotoxocity in cultured myocardial cells by the increase of beating rate as well as th decrease of lipid peroxidation. From these results, they show that XO/HX is cardiotoxic in cultured myocardial cells derived from neonatal rat, and it suggests that TRS, FT, PT, CA and PRR extracts are positively effective in the blocking in XO/HX-induced cardiotoxicity.

• 사상체질의학회지
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• 제14권1호
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• pp.67-78
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• 2002

Jowiseungcheongtang(JST) has been in Sasang constitution medicine for many years as a therapeutic agent for cerebral disease. But the effect of Jowiseungcheongtang(JST) on neurotoxicity is not known. The purpose of this study was to determine the effects of Jowiseungcheongtang(JST) on the hippocampal cell injured by Xanthine Oxidase/Hypoxanthine. The results were as follows: 1. XO/HX decreased the survival rate of the cultured hippocampal cells on NR assay and MTT assay. 2. JST water extract have efficacy of decreasing a amount of lipid peroxidation increased by XO/HX in cultured hippocampal cells. 3. JST water extract have efficacy of increasing DNA synthesis decreased by XO/HX in cultured hippocampal cells. From the above results, It is concluded that JST has marked efficacy in preventing cultured hippocampal cells from the damages by XO/HX.

• 대한약리학회지
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• 제27권2호
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• pp.125-133
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• 1991

내피세포가 제거된 토끼의 적출 장간막 동맥에서 토끼의 대동맥 내피세포로 부터 A23187과 acetylcholine은 NO와 유사한 혈관 이완성 물질 (EDRF)을 유리한다. 이에 첨가하여 A23187은 acetylcholine과는 달리 superoxide anion에 의하여 파괴되지 않는 EDRF도 유리시킴을 확인하고 이의 특성에 대하여 연구하였다. 정상적인 생리영양액에서는 A23187과 acetylcholine의 용량-반응 곡선은 내피세포에 의존하지 않는 sodium nitroprusside의 곡선과 유사하였다. 이들은 methylene blue에 의하여 억제되었다. Hypoxanthine (HX)과 xanthine oxidase (XO)를 bath내로 투여시 phenylephrine에 의한 수축이 일과성으로 증가한 후 지속적으로 이완되었다. HX-XO 반응중에는 A23187은 장간막 동맥을 즉각적으로 이완시켰으나 acetylcholine의 이완작용은 소실되었다. A23187에 의하여 야기되는 장간막동맥의 이완은 50 mM $K^+-PSS$로 수축을 야기시켰을 때에는 나타나지 아니하였다. Superoxide dismutase를 전처치하였을 때는 HX-XO 반응중에도 acetylcholine 뿐만아니라 A23187에 의한 장간막 동맥의 수축은 이완되었다. 한편, acetylcholine에 의하여 야기되는 장간막 동맥의 이완은 A23187에 의하여 야기되는 이완에 비하여 phorbol 12-myristate 13-acetate (PMA)에 훨씬 더 민감하게 억제되었다. 내피세포의 기능과는 무관한 sodium nitroprusside에 의하여 야기되는 이완은 PMA에 의하여 영향을 받지 아니하였다. 이상의 결과로 미루어 볼때, A23187과 acetylcholine은 methylene blue에 의하여 억제되는 내피세포 의존성 이완을 야기시키고, 첨가하여 A23187은 어떤 병적 환경 아래서는 superoxide anion과 PMA에 저항성을 지닌 혈관이완성 물질을 유리하는 것으로 사료된다. 앞으로 A23187에 의하여 유리되는 혈관 이완성 물질이 superoxide anion에 의존하여 생성된 것인지에 대하여는 추후의 연구과제이다.

• 약학회지
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• 제41권1호
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• pp.99-104
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• 1997

In order to elucidate the cytotoxic effect of oxygen radicals on cultured spinal dorsal root ganglion(DRG) neurons derived from mouse. the neurotoxic effect of oxygen radicals w as examined after cultured DRG neurons were exposed to xanthine oxidase(XO) and hypoxanthine(HX)-oxygen radical generating system. In addition. neuroprotective effect of epidermal growth factor(EGF) against oxidant-induced neurotoxicity was also evaluated in these cultures. The results were, as follows: 1. Lethal concentration 50(LC$_{50}$) was 35mU/ml XO and 0.1mM HX in cultured DRG neurons. 2. Oxygen radicals induced the morphological changes such as the decrease of cell number and loss of neurites in these cultures. 3. EGF increased the cell viability and neurofilament in neurons damaged by oxygen radicals. From above the results, it is suggested that oxygen radicals have a cytotoxic effect on cultured DRG neurons of neonatal mouse and selective neurotrophic factors such as EGF are, effective, in blocking the neurotoxicity induced by oxygen radicals in cultured spinal DRG neurons.

• 대한한방내과학회지
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• 제21권2호
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• pp.283-290
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• 2000

The purpose of this study is to examine the toxic effects caused by xanthine oxidase/hypoxanthine(XO/HX) and the effects of herbal extracts such as Jingansikpungtang(JST) and Gamijingansikpungtang(GJST) on the treatment of the toxic effects. For this purpose, experiments with the cultured nerve cells from the spinal motor neurons of new born mice were done. The results of these experiments were as follows. XO/HX, a oxygen radical-generating system, decreased the survival rate of the cultured cells on NR assay. MTT assay, the amount of neurofilaments and increased the amount of total proteinand increased the lipid peroxidation and the amount of LDH JST has the efficacy of increasing the amount of neurofilaments and total protein, and decreasing the lipid peroxidation and the amount of LDH, GJST has efficacy of increasing the amount of neurofilaments and total protein, and decreasing lipid peroxidation and the amount of LDH. From the above results, it is concluded that JST and GJST have marked efficacy as a treatment for the damages caused in the XO/HX mediated oxidative stress. And JST and GJST are thought to have certain pharmacologicall effects. Further clinical study of this pharmacological effects of JST and GJST should be complemented.

• 동의생리병리학회지
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• 제16권5호
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• pp.989-1000
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• 2002

The purpose of this study is to examine the toxic effects caused by xanthine oxidase/hypoxanthine(XO/HX) and the effects of herbal extracts such as JHYJ and GJHYJ on the treatment of the toxic effects. For this purpose, experiments with the cultured hippocampal cells from new born mice were done. The results of these experiments were as follows. 1. XO/HX, a oxygen radical-generating system, decreased the survival rates of the cultured cells on XTT assay and INT assay, the amount of DNA syntheses, and the amount of neurofilaments, and increased the lipid peroxidation. 2. JHYJ and GJHYJ have the efficacy of increasing the survival rates of the cultured cells. 3. JHYJ and GJHYJ have the efficacy of increasing the amount of neurofilaments and of decreasing the lipid peroxidation. 4. JHYJ and GJHYJ have the efficacy of increasing the amount of DNA syntheses. From the above results, it is suggested that Jihwangyeumja and Gamijihwangyeumja have marked efficacy as a treatment for the damages caused by the XO/HX-mediated oxidative stress. And Jihwangyeumja and Gamijihwangyeumja are thought to have certain pharmacological effects. Further dinical study of this pharmacological effects of Jihwangyeumja and Gamijihwangyeumja should be complemented.

• 대한의생명과학회지
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• 제11권3호
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• pp.319-326
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• 2005

It is well known that oxidative stress of reactive oxygen species (ROS) may be a causative factor in the pathenogenesis of bone disorder on osteoblast or osteoclast. The purpose of this study was to evaluate the cytotoxicity of oxidative stress, protective effect of glutamate receptor antagoinst against ROS-induced osteotoxicity, secretion of tumor necrosis factor $(TNF)-\alpha$ and the expression of c-fos gene in the cultured rat osteoblasts and osteoclasts. Cell viability by MTS assay or !NT assay, activity of glutathione peroxidase (GPx), lipid peroxidation (LPO) activity, protein synthesis by sulforhodamine B (SRB) assay, alkaline phosphatase (ALP) activity, lactate dehydrogenase (LDH) activity, MTS assay for NMDA (N-methyl-D-aspartate) receptor antagonist or AMPA/kainate receptor antagonist, measurement for $TNF-\alpha$, and c-fos gene expression were performed after these cells were treated with or without various cocentrations of xanthine oxidase (XO), hypoxanthine (HX), D-2-amino-5-phosphonovaleric acid (APV), 7-chlorokynurenic acid (CKA), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and 6,7-dinitroquinoxaline-2,3-dione (DNQX), respectively. In this study, XO/HX showed decreased cell viability and glutathione peroxidase (GPx) activity, but it showed increased LPO activity, $TNF-\alpha$ secretion and c-fos expression. APV and CKA incresed protein sythesis and ALP activity. While, CNQX or DNQX did not show any protective effect in LDH activity or cell viability. From these results, XO/HX showed cytotoxic effect in cultured rat osteoblast or osteoclast, and also NMDA receptor antagonist such as APV or CKA was effective in blocking XO/HX-induced osteotoxicity in these cultures.

• 동의생리병리학회지
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• 제17권4호
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• pp.996-1001
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• 2003

It is well known that oxidative stress of reactive oxygen species(ROS) may be a causative factor in the pathogenesis of bone disorder. The purpose of this study was to evaluate the cytotoxicity of oxidative stress. Cell viability by MTS assay or INT assay, activity of glutathione peroxidase(GPx), lipid peroxidation(LPO) activity and cell viablity. And also protctive effect of glutamate receptors against ROS-induced osteotoxicity was examined by protein synthesis, alkaline phosphatase (ALP) activity and lactate dehydrogenase (LDH) activity in cultured rat osteoblasts and osteoclasts. XO/HX decreased cell viability and GPx activity, protein synthesis and ALP activity, but increased LPO activity and LDH activity. In the protective effect, N-methyl-D-aspartate (NMDA) receptor antagonists or AMPA/kainate receptor antagonists such as D-2-amino-5-phosphonovaleric acid (APV), 7-chlorokynurenic acid (CKA), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and 6,7-dinitroquinoxaline-2,3-dione (DNQX), NMDA receptor antagonists but AMPA/kainate receptor antagonists showed protective effect on xanthine oxidase (XO) and hypoxanthine (HX) in these cultures by the increse of protein synthesis, ALP activity.

• 대한한의학방제학회지
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• 제7권1호
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• pp.131-141
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• 1999

In order to eludidate the mechanism of oxidative stress in cultured spinal motor neurons damaged by oxygen free radicals, cytoxicity was assesed by MTT assay and NR assay after spinal motor neurons from mouse were cultured in media containing various concentrations of xanthine oxidase(XO) and hypoxanthine(HX) for 3 hours. In addition, neuroprotective effects of several herb extracts on oxidant-induced neurotoxicity were examined in these cultures, compared with nerve growth factors such as basic fibroblast growth factor(bFGF). XO/HX decreased cell viability in dose- and time dependent manners on cultured mouse spinal motor neurons, and MTT50 and NR50 values were measured at 20mU/ml XO and 0.1mM HX for 3 hours in these cultures. bFGF significantlt increased cell viability. In neuroprotective of herb extracts, Epimedium Koreanum Nakai(EK) and Alpinia oxphylla Mig(IJI) was very effective in the prevention of the neurotoxicity induced by XO/HX in cultured mouse spinal motor neurons. From the above results, it is suggested that XO/HX shows toxic effect in cultured mouse spinal motor neurons and selective herb extracts such as Epimedium Koreanum Nakai(EK) and Alpinia oxphylla Mig(IJI) were very effective in the increase of cell viability against the neurotoxicity induced by oxygen radicals in these cultures.