Kainic Acid에 의한 흰쥐 해마 신경원 가지돌기의 독성변화에 대한 광학 및 전자현미경 연구

최진영,김경용,이원복,김동창 대한해부학회 1993 Anatomy & Cell Biology Vol.26 No.4

열다한소탕(熱多寒少湯)이 kainic acid에 의해 유발된 mouse의 해마체 손상에 미치는 영향

김일환,김경요,Kim, Il-hwan,Kim, Kyung-yo 사상체질의학회 1999 사상체질의학회지 Vol.11 No.2

1. 연구목적 kainic acid를 실험동물에 주입할 경우 간질발작을 일으키고 변연계 특히 해마체 부위에서 조직의 손상이 일어나게 되는데 이는 사람에 있어 측두엽성 간질에서 보이는 구조적 변화와 유사한 것이다. 본 실험은 신경독성 물질인 kainic acid로 마우스의 해마체에 손상을 유발하고 이 경우에 열다한소탕이 신경보호효과가 있는지의 여부를 알고자한 것이다. 2. 연구방법 kainic acid를 경구투여하고 열다한소탕을 3주일간 복용시키면서 각각 1, 2, 3일과 1주, 3주에 조직을 관찰하였다. 조직손상은 해마체의 CA1, CA3과 thalamus, amygdala 등에서 c-fos 와 DNA fragmentation의 출현 율로 지표를 삼았으며 광학현미경하에서 육안적 관찰을 병행하였다. 3. 결과 및 결론 kainic acid만을 투여한 대조 군에서는 실험 3주째에도 손상지표인 c-fos 와 DNA fragmentation이 발견되었으나 열다한소탕을 투여한 실험 군에서는 실험 약 3일 째부터 손상지표의 발현이 줄어 7일째에는 나타나지 않음을 알 수 있었다. 또한 현미경 하의 육안적 관찰에서도 실험초기 많은 손상을 보였던 신경세포가 2주일 후 어느 정도 회복되는 것을 볼 수 있었다. 이로써 열다한소탕이 kainic acid로 유발된 실험동물의 해마체손상에 대하여 신경세포 보호 효과 및 손상 억제 효과가 있음을 알 수 있었다. 1. Purpose : Systemic injection of kainic acid in experimental animals induces the limbic seizure and structural damages in hippocampus and amygdala which resembles the changes in human temporal lobe epilepsy. The author performed this study to investigate the neuroprotective effects of Yuldahansotang, on the neurotoxicity induced by kainic acid in the hippocampus in rats. 2. Method : Kainic acid was administered intraperitoneally. And feeding with Yuldahansotang for 3 weeks after kainic acid administration. Seizure were induced in male mice (kainate 10-40 mg/kg i.p) and animals were sacrified at various time-points after injection. The experimental animals were sacrificed at 1, 2, 3day and 1, 3weeks while Yuldahansotang administrations. Seizure were graded using a behavioral scale developed in our laboratory. c-fos belong to immediate early genes(IEGs) known to have rapid and brief responses. And neuronal injury was assayed by examining DNA fragmentation using in situ nick translation histochemistry. 3. Results & Conclusion : Seizure severity paralled kainate dosage. At higher doses DNA fragmentation is seen mainly in hippocampus in area CA3, and variable in CA1, thalamus, amygdala within 24 h, is maximal within 72 h, and is large gene by 7 days after administration of kainate. And we can't see the expression of DNA fragmentation and c-fos in the mice what feeded by Yuldahansotang after 7 days from kainic acid administration. It is consequently suggested that Yuldahansotang may attenuate the kainic acid-induced neuronal degeneration and increase the immunoreactivity of hippocampus in mouse.

Kainic acid로 유발된 seizure가 흰쥐 소뇌의 GABA와 GABA 수송체에 미치는 영향

홍혜남,이종환,임진옥,황승준,김동호 대한해부학회 1999 Anatomy & Cell Biology Vol.32 No.6

A comparative study of the phenotype with kainic acid-induced seizure in DBA/2 mice from three different sources

강경구,김영인,서민수,성수은,최주희,이시준,정영석,조준영,황대연,박상준,김길수 한국실험동물학회 2020 Laboratory Animal Research Vol.36 No.4

The kainic acid-induced seizure mouse model is widely used in epilepsy research. In this study, we applied kainic acid to the subcutaneous injections of three different sources of DBA/2 mice to compare and evaluate the seizure response. The three mouse sources consisted of DBA/2Kor1 (Korea FDA source), DBA/2A (USA source), and DBA/2 (Japan source), and were purchased from different vendors. To compare the responses of DBA/2 mice to kainic acid injections, we examined the survival rate, seizure phenotype scoring, and behavioral changes. We also evaluated brain lesions using histopathological analysis. Following the administration of kainic acid, almost half of the cohort survived, and the seizure phenotype displayed a moderate level of sensitivity (2 ~ 4 out of 6). In the histopathologic analysis, there was no change in morphological features, and levels of glial fibrillary acidic protein (GFAP) and ionized calcium binding adaptor molecule 1 (Iba-1) increased in the kainic acid-treated groups. However, there was no difference in the neuronal nuclei (NeuN) expression level. All the data showed that the responses in the kainic acid-treated group were similar across the three strains. In conclusion, our results suggest that the three sources of DBA/2 mice (DBA/2Kor1, DBA/2A, and DBA/2B) have similar pathological responses to kainic acid-induced seizures.

Neuroprotective Effects of Butterbur and Rough Aster Aainst Kainic Acid-Induced Oxidative Stress in Mice

Oh, Sang-Hee,Sok, Dai-Eun,Kim, Mee-Ree 충남대학교 암공동연구소 2006 암공동연구소 업적집 Vol.5 No.

The separate and combined ncuroprotective effects of rough aster (Aster scaber) and butterbur (Petasite japonicus) extracts against oxidative damage in the brain of mice challenged with kainic acid were examined by comparing behavioral changes and biochemical parameters of oxidative stress. Rough aster butanol extract (400 mg/kg) and/or butterbur butanol extract (150 or 400 mg/kg) were administered to male [CR mice, 68 weeks old, through a gavage for 4 days consecutively, and on day 4, kainic acid (50 mg/kg) was administered intrapcritoneally. Compared with the vehicle-treated control, no significant changes in body and brain weight were observed in mice administered rough aster or butterbur butanol extract. Administration of kainic acid only, causing a lethality of approximately 54%, resulted in a significant decrease of total glutathione level and increase of thiobarbituric acid-reactive substances (TBARS) value in brain tissue. The administration of butterbur or rough aster extract (400 mg/kg) decreased the lethality (50%) of kainic acid to 25%, alleviated the behavioral signs of neurotoxicity, restored the cytosolic glutathione level of brain homogenate to approximately 80% (P<.05), and reduced kainic acid-induced increases in TBARS values. In contrast to no significant neuroprotection by butterbur extract at a low dose (150 mg/kg), the combination of rough aster extract and butterbur extract reduced the lethality to 12.5%. Moreover, the combination delayed the onset time of behavioral signs by twofold, and significantly preserved the level of cytosolic glutathione peroxidase and glutathione reductasc activities. However, the other biochemical parameters were not altered significantly by the combination. Thus, the combination of two vegetable extracts significantly increased the neuroprotective action against kainic acid-induced neurotoxicity. Based on these findings, the combination of butterbur extract and rough aster extract contains a functional agent or agents that protect against oxidative stress in the brain of mice.

Neuroprotective Effects of Butterbur and Rough Aster Aainst Kainic Acid-Induced Oxidative Stress in Mice

Oh, Sang-Hee,Sok, Dai-Eun,Kim, Mee-Ree 충남대학교 암연구소 2006 암연구소 업적집 Vol.5 No.-

The separate and combined ncuroprotective effects of rough aster (Aster scaber) and butterbur (Petasite japonicus) extracts against oxidative damage in the brain of mice challenged with kainic acid were examined by comparing behavioral changes and biochemical parameters of oxidative stress. Rough aster butanol extract (400 mg/kg) and/or butterbur butanol extract (150 or 400 mg/kg) were administered to male [CR mice, 68 weeks old, through a gavage for 4 days consecutively, and on day 4, kainic acid (50 mg/kg) was administered intrapcritoneally. Compared with the vehicle-treated control, no significant changes in body and brain weight were observed in mice administered rough aster or butterbur butanol extract. Administration of kainic acid only, causing a lethality of approximately 54%, resulted in a significant decrease of total glutathione level and increase of thiobarbituric acid-reactive substances (TBARS) value in brain tissue. The administration of butterbur or rough aster extract (400 mg/kg) decreased the lethality (50%) of kainic acid to 25%, alleviated the behavioral signs of neurotoxicity, restored the cytosolic glutathione level of brain homogenate to approximately 80% (P<.05), and reduced kainic acid-induced increases in TBARS values. In contrast to no significant neuroprotection by butterbur extract at a low dose (150 mg/kg), the combination of rough aster extract and butterbur extract reduced the lethality to 12.5%. Moreover, the combination delayed the onset time of behavioral signs by twofold, and significantly preserved the level of cytosolic glutathione peroxidase and glutathione reductasc activities. However, the other biochemical parameters were not altered significantly by the combination. Thus, the combination of two vegetable extracts significantly increased the neuroprotective action against kainic acid-induced neurotoxicity. Based on these findings, the combination of butterbur extract and rough aster extract contains a functional agent or agents that protect against oxidative stress in the brain of mice.

Kainic Acid 투여로 경련이 유도된 백서에서 Propofol의 항경련 효과

김종욱(Jong Uk Kim),이은호(Eun Ho Lee),박평환(Pyung Hwan Park),한성민(Sung Min Han),이종환(Jong Whan Lee),홍혜남(Hea Nam Hong),황승준(Seung Jun Hwang) 대한마취과학회 1999 영문부록 Vol.- No.-

Background : Propofol has been used to treat status epilepticus, but anticonvulsant effects of propofol were not proven distinctly. We examined the effect of propofol on seizure induced by kainic acid that is a glutamate analogue with excitotoxic properties. Methods : All experimental rats were injected with kainic acid (10 mg/kg, i.p.). When the salivation and focal convulsion were observed, propofol (50 mg/kg, n=8) or saline (5 ml/kg, n=8) was injected intraperitoneally. The seizure activities were observed for 6 hrs. Acid fuchsin stain and immunoreactivity (heat-shock protein) have been used as indicators of injured neurons and epileptic insults. Results : There were significant decrease of seizure activity in propofol treated group (p<0.01). Conspicuous cytopathologic reactions were evident in saline treated group but no detectable histologic change in propofol group. Immunocytochemical reactions with a monoclonal antibody against HSP-70 were evident in the saline group but no reaction in propofol group. Conclusions : These results indicate that propofol has marked anticonvulsant effects against seizure induced by kainic acid in the rat. (Korean J Anesthesiol 1999; 36: 1038∼1045)

흰쥐 해마에서 경련에 의해 발현 유도된 MKP-1에 의한 MAPK 활성 조절

유범희,강웅구,안용민,정선주,전송희,박주배,김용식 大韓神經精神醫學會 1999 신경정신의학 Vol.38 No.4

연구목적 : 전기경련 충격(Electroconvulsive shock, ECS) 및 카이닌산(kainic acid)에 의한 경련은 흰쥐 해마에서 mitogen activated protein kinase(MAPK)를 활성화시키며, 동시에 MAPK 불활성화 효소인 MAPK phosphatase-1(MKP-1)의 발현을 일으킨다. 이 연구의 목적은 경련에 의해 발현된 MKP-1이 역시 경련에 의해 활성화된 MAPK의 불활성화에 관여하는지를 알아보고자 하는 것이다. 방 법 : 흰쥐에 ECS를 가하여 해마에서 MKP-1 발현을 일으킨 뒤 다시 ECS를 가하고, 두 번째 ECS에 의한 MAPK의 일시적 활성화가 MKP-1의 발현상태에 의해 영향받는지를 알아보았다. 또한 흰쥐에 지속적인 MAPK 활성화 및 MKP-1 발현을 일으키는 카이닌산을 투여한 뒤 해마에서 MKP-1의 발현과 MAPK 활성과의 관계를 알아보았다. 결 과 : ECS후 해마에서 타이로신 인산화 면역블롯 및 효소활성으로 측정한 MAPK의 인산화 및 활성은 MKP-1의 발현이 일어나 있는 경우와 그렇지 않은 경우 사이에 차이를 보이지 않았다. 카이닌산의 투여에 의해 MAPK 활성화가 일어나는 경우, 뒤이어 MKP-1 발현이 일어나지만, 이렇게 발현된 MKP-1은 MAPK 활성을 충분히 감소시키지 못하였다. 결 론 : MKP-1은 흰쥐 해마에서 ECS 및 카이닌산에 의한 MAPK의 활성화를 차단하는데 큰 역할을 하지 않는다. Objectives : Both electroconvulsive shock(ECS)- and kainic acid-induced seizures activate mitogen-activated protein kinases(MAPKs) in rat hippocampus. They can also induce the expression of MAPK phosphatase-1(MKP-1) in rat hippocampus. MKP-1 is known as a specific MAPK deactivator. This study aimed to elucidate the role of MKP-1 in the deactivation of MAPKs in rat hippocampus. Methods : In order to induce MKP-1 in the hippocampus, ECS was given to the rats. At the time points when MKP-1 was sufficiently induced, the second ECS was given to them and the subsequent phosphorylation or activation of MAPKs were measured in the hippocampus. A second group of rats were injected with kainic acid and the relationship between MKP-1 expression and MAPK phosphorylation was examined in their hippocampi. Results : The expression of MKP-1 did not influence the phosphorylation or activation of MAPKs following ECS in rat hippocampus. Kainic acid-induced expression of MKP-1 did not significantly reduce the phosphorylation of MAPKs. Conclusion : MKP-1 did not play a significant role in the deactivation of MAPKs which were activated by ECS or kainic acid in rat hippocampus.

백서 해마에서 카이닌산 유도 경련에 의한 JNK 경유 신호전달경로 활성화의 발달 단계에 따른 변화

김종흔,정희연,노명선,안용민,강웅구,김용식,조수철 大韓神經精神醫學會 2001 신경정신의학 Vol.40 No.5

연구목적 : 어린 백서의 해마에서 전기경련충격(electroconvulsive shock, ECS) 후 활성화되는 것으로 알려져 있는 MAPK계 신호전달경로가 카이닌산(Kainic acid, KA)에 의한 경련 후 어떤 양상으로 활성화되는 지를 초기 발달 단계에 따라서 관찰하고자 하였다. 방 법 : 생후 7일, 14일, 21일 된 어린 백서와 성년 백서의 해마에서 면역블롯 방법으로 MAPK계 효소의 인산화 효소 중 하나인 SAPK/ERK kinase-1(SEK-1), MAPK계 효소인 c-Jun N terminal protein kinase(JNK), 전사인자 c-Jun 및 조기유전자 단백질 Fos의 단백질 양과 기저 인산화를 측정하였고, 백서에게 KA를 주사하여 경련을 유발한 후 위 단백질들의 인산화 변화의 양상을 같은 방법으로 관찰하였다. 결 과 : SEK-1, JNK, c-Jun 모두 연령 증가에 따른 단백질 양 자체에는 변화가 없었다. JNK와 c-Jun은 기저 인산화의 양적 변화도 없었으나 SEK-1의 경우 기저 인산화 양이 생후 7일에 가장 높고 이후 연령이 높아짐에 따라 감소하였다. JNK의 경우 KA 주사 후 인산화 변화 양상이 관찰되지 않았으나 SEK-1과 c-Jun은 생후 14일부터 연령의 증가와 함께 인산화 양도 증가하였다. Fos 역시 생후 7일부터 발현되어 연령의 증가와 함께 발현양이 증가하였다. 결 론 : 백서 해마에서의 MAPK계 신호전달계의 성숙은 연령의 증가에 따라, 또 신호전달 물질에 따라 차이를 보였다. KA 유도 경련 후 JNK는 생후 21일까지도 활성화가 되지 않는 것으로 보아, 세포주에서 알려진 SEK-1 - JNK-c-Jun - Fos로 연결되는 신호전달 경로가 어린 백서의 해마에서는 적용될 수 없음을 시사한다. Objective : We observed the developmental pattern of activation of MAPK signal transduction pathways known to be activated by eletroconvulsive shock(ECS) in young rat hippocampus after kainic acid(KA)-induced seizure. Methods : We used the method of immunoblotting for examining the basal protein amount and basal level of phosphorylation of MAPK kinase(SAPK/ERK kinase -1, SEK-1), MAPK(c-Jun N terminal protein kinase, JNK), transcription factor(c-Jun) and immediate early gene proteins(Fos) in rat hippocampus at postnatal day 7, 14 and 21, respectively. We also examined the changes of phosphorylation of those proteins after kainic acid-induced seizure in the same way. Results : The basal protein amounts of SEK-1, JNK, and c-Jun did not show age-dependent changes and basal level of phosphorylation of JNK and c-Jun remains unchanged throughout the early developmental period. The basal level of phosphorylation of SEK-1 was peaked at postnatal 7 days and then decreased with aging. After kainic acid-induced seizure, the change of phosphorylation of JNK was not observed but those of SEK-1 and c-Jun increased after postnatal day 14. The expression of Fos was observed at postnatal day 7 and also increased with aging. Conclusion : These results show that the MAPK signal transduction system in rat hippocampus matures in accordance with aging, but the process of maturation differs depending specific proteins. This study suggests the signal transduction cascade(SEK-1 - JNK - c-Jun - Fos) which is well established in cell line studies may not be applied to rat hipposcampus because we could not observe the activation of JNK after KA-induced seizure in young rat hippocampus.

Kainic Acid로 유발된 경련이 흰쥐 대뇌피질의 Vasoactive Instestinal Polypeptide 신경세포에 미치는 영향에 관한 면역조직화학적 연구

허영범,변우현,이원규,김정혜,안희경 대한해부학회 1995 Anatomy & Cell Biology Vol.28 No.2