Pilocarpine에 의한 경련과 기억력 저하에 Cycloheximide와 Dexamethasone이 미치는 영향

김용민,박정숙,한형수 慶北大學校 醫科大學 1996 慶北醫大誌 Vol.37 No.3

목적 : Cycloheximide 또는 dexanethasone이 pilocarpine으로 유발된 경련의 발생과 뇌조직 손상으로 인한 기억력 저하에 어떤 영향을 미치는지 연구하였다. 대상 및 방법 : 생쥐에 pilocarpine(250 ㎎/㎏)을 투여하여 야기된 경련 및 기억장애가 10㎎/㎏의 cycloheximide 또는 1.5㎎/㎏ 의 dexamethasone을 전처치하였을 때 나타나는 작용을 비교하였다. 기억력검사는 Morris water maze방법을 이용하였다. 결과 : Dexamethsone 또는 cycloheximide의 전처치로서 pitocarpine에 의한 경련 시작 시간이나 경련의 강도에 영향을 주지 않았다. Pilocarpine 투여 17 일 후 기억력을 검사한 결과, reference memory을 working memory 장애가 야기되었으나, cycloheximide 및dexamethasone은 pilocarpine 투여 7일후 4일간 투여하였을 때 cycloheximide만이 pilocarpine에 의해 기억력이 저하되는 것을 억제하였다. 결론 : Cycloheximide는 항경련 작용 없이도 pilocarpine에 의한 기억력 저하를 억제할 수 있는 것으로 미루어 경련과 기억력저하의 유발과정이 다를 수 있는 것으로 보여진다. The effect of cycloheximide, inhibitor of protein synthesis, and dexamethasone, synthetic glucocorticoid, on the genesis of pilocarpine-induced seizure and seizure-related memory deficit was observed in mice. Twenty days after pilocarpine(250 ㎎/㎏) administration showed memory deficit. Cycloheximide (10㎎/㎏/day) and dexamethasone (1.5 ㎎/㎏/day) were given once a day during 7 to 10 days following pilocarpine treatment. Pretreatment with cycloheximide or dexamethasone did not influence on the onset of seizure and behavioral seizure activity induced by pilocarpine. In contrast to dexamethasone, cycloheximide treatment inhibited pilocarpine-induced memory defect on the basis of reference and working memory version of Morris water maze. These results show that cycloheximide, not dexmethsone, can reduce pilocarpine-induced memory deficit without anticonvulsant property.

Brief exposure to cycloheximide prior to electrical activation improves in vitro blastocyst development of porcine pathenogenetic and reconstructed embryos

Naruse, K.,Quan, Y.S.,Kim, B.C.,Lee, J.H.,Park, C.S.,Jin, D.I. 충남대학교 형질전환복제돼지연구센터 2007 논문집 Vol. No.10

To investigate the effects of cycloheximide exposure before electrical activation of in vitro-matured porcine oocytes on the subsequent development of parthenogenetic embryos, cumulus-free mature oocytes were exposed to NCSU-23 medium containing cycloheximide (10μg/mL) for 0, 5, 10, 20, 30 and 60 min, activated by electrical pulse treatment (1.5 kV/cm, 100μs) and then cultured in PZM-3 for 7 days. To evaluate the effects of cycloheximide on the activation of nuclear transfer embryos, reconstructed embryos were electrically activated by two DC pulses (1.2 kV/cm, 30μs) before or after exposure to cycloheximide. The reconstructed embryos were allocated into four groups: electrical pulse treatment alone (Ele); exposure to cycloheximide for 10 min followed by electrical activation (CHX + Ele); electrical activation followed by exposure to cycloheximide for 6 h (Ele + CHX); exposure to cycloheximide for 10 min, followed by electrical activation and a further exposure to cycloheximide for 6 h (CHX + Ele + CHX). The activated reconstructed embryos were cultured in PZM-3 for 6 days. Oocytes treated with 10 min exposure to cycloheximide followed by electrical activation had a significantly higher percentage of blastocyst formation compared to control oocytes and oocytes exposed for≥30 min. In the reconstructed embryos, the blastocyst development rates of embryos exposed to cycloheximide (CHX + Ele, Ele + CHX and CHX + Ele + CHX) were significantly higher than those of the control group (Ele). Among the cycloheximide-treated groups, the CHX + Ele group had increased development rate and total blastocyst cell number, though these values were not significantly different from those observed in the other cycloheximide-treated groups. To evaluate the quality of NT embryos treated with cycloheximide, apoptosis in blastocysts was analyzed by TUNEL assay. The 10 min exposure to cycloheximide prior to electrical activation significantly reduced cell death compared with longer exposure to cycloheximide after electrical fusion. In conclusion, brief exposure to cycloheximide prior to electrical activation may increase the subsequent blastocyst development rates in porcine parthenogenetic and reconstructed embryos.

Isolation and Physico-Chemical Properties of Cycloheximide-isomer from Streptomyces sp.NA-52

Yoon,Won-Ho,Lim,Dae-Seog,Kim,Chang-Han,Yoon,Sang-Kun 건국대학교 동물자원연구센터 1999 動物資源硏究誌 Vol.20 No.-

새로운 분리균주 Streptomyces sp. NA-52는 Pyricularia oryzae와 Chlorella regularis에 대해 항균활성물질을 생산하는 것으로 밝혀졌다. 그 균주에 의해 생산되는 항생물질 복합물 중 (compound A 및 B), 활성이 좋은 화합물 A가 분리되었으며, 그것의 이화학적 성질이 조사되었다. 그 화합물의 분자량은 281(EI-MS)로 결정되었고, UV, IR 및 ¹H, 13C NMR 스펙트럼 분석결과, cycloheximide(4-[2-(3, 5-dimethyl-2-oxocyclohexyl)-2-hydroxyethyl]-2,6-piperidinedione)의 이성체와 같은 glutarimide 그룹의 항생물질로 분류되었다. 한편, 백자무우 씨앗의 발아시험결과, compound A는 cycloheximide와 마찬가지로 백자무우씨앗에 대해 강한 독성을 가지고 있었으나, compound B는 그 씨앗을 100% 발아시켰다. 지금까지의 cycloheximide 이성체와 비교해 볼 때, compound B는 백자무우씨앗에 대해 전혀 독성을 나타내지 않는, glutarimide 그룹내의 새로운 cycloheximide 이성체로 추정되었다. 결과적으로 cycloheximide와 같은 glutarimide 그룹의 항생물질이 Streptomyces griseus가 아닌 다른 방선균종에서도 생산된다는 것으로 밝혀졌다. A new isolate of Streptomyces sp. NA-52 was found to produce antibiotics with antimicrobial spectra against Pyricularia oryzae and Chlorella regularis. The main active component, compound A, of the antibiotic complex (compounds A and B) produced by the strain was isolated, and its physico-chemical properties were investigated. The molecular weight of the compound was determined to be 281 (EI-MS). UV, 1R, and ¹H and 13C NMR spectra suggested that the compound is a cycloheximide (4-[2-(3,5-Dimethyl-2-oxocyclohexyl)-2-hydroxyethyl]-2,6-piperidinedione) isomerlike glutarimide group antibiotic. Ina germination test using BAIKJA radish seed, neither compound A nor cycloheximide germinated the seeds, but compound B completely germinated them. Compared with known cycloheximide-isomers, compound B may be thought of as a new cycloheximide-isomer, without toxicity for BAIKJA radish seed, in the glutarimide group.

간세포 Apoptosis에 대한 형태학적 연구

한치동,조창호,서인수 慶北大學校 醫科大學 1995 慶北醫大誌 Vol.36 No.4

목적 : 본 연구는 간세포에 괴사를 주로 일으키는 물질로 알려진 사염화탄소와 apoptosis를 일으키는 것으로 알려진 cycloheximide로 간세포에 일어난 괴사와 apoptosis의 차이를 형태학적으로 알아보고 또 최근 개발된 in situ nick translation방법을 이용한 ApopTag염색을 통해 apoptosis를 쉽게 찾을 수 있는가를 알아보기 위해 시도하였다. 대상 및 방법 : 표준식으로 사육한 쥐에 cycloheximide를 체중 1㎏당 1.5㎎ 또는 50㎎이 되도록 정맥내에 주사하였고, 다른 동물은 사염화탄소를 체중 1㎏당 1㎎씩 복강내로 주사하였다. Cycloheximide 주사 후 30분에서 6시간 사이에, 사염화탄소 주사 후는 12시간에서 24시간 사이에 도살하여 H-E 및 ApopTag로 염색하에 apoptosis수를 계수하였고, 전자현미경으로도 관찰하였다. 결과 : Cycloheximide군은 H-E염색상 1.5㎎/㎏군에서는 30분후 100개의 간세포당 0.82개, 6시간후에 4.29개의 apoptosis가 나타났고 50㎎/㎏투여군에서는 처음 1.85개에서 6시간 후 9.86개가 나타나 용량의존성을 보였다. 사염화탄소 군은 H-E 염색에서 처음 0.46개에서 후에 1.88개로 증가하였다. ApopTag염색에서는 cycloheximide 투여군에서는 10-15%의 검출률이 더 높았으나 사염화탄소 투여군에서는 2-3배의 검출률을 보여 상반된 결과를 보였다. 결론 : 이상의 성적으로 보아 ApopTag는 apoptosis의 쉬운 검출을 위해 매우 유용한 것으로 생각되나 괴사 세포에 양성을 보일 수도 있으므로 결과해석에는 신중한 검토가 필요할 것으로 생각된다. It is well known that there are two ways of cell death; necrosis and apoptosis. Recently apoptosis is vigrously studied in concern with mechanism of hyperplasia and tumor promotion. This study was carried out to compare the morphologic differences between necrosis and apoptosis and to verify the usefulness of recently developed ApopTag for the detection of apoptotic cells. Necrosis was induced by carbon tetrachloride and apoptosis was induced by cycloheximide. Rats were received intravenous single dose of 1.5㎎/㎏ or 50㎎/㎏ of cycloheximide and sacrificed 30 minutes, 1 hour, 3 hours, and 6 hours after injection, other rats were received intraperitoneal single does of 1㎖ of carbon tetrachloride and sacrificed 12, 15 and 24 hours after injection. Numbers of apoptotic cells were counted in hematoxylin and eosin and ApopTag stained tissue. Apoptosis was also observed under electron microscope. Cycloheximide 1.5㎎/㎏ group showed 0.82 apoptotic cells among 100 liver cell nuclei after 30minutes; 2.19 after 1 hour; 2.64 after 3 hours; 4.29 after 6 hours. Cycloheximide 5㎎/㎏ group showed 1.85 apoptotic cells among 100 liver cell nuclei after 30 minutes; 5.42 after 1 hour; 9.69 after 3 hours; 9.86 after 6 hours respectively. Apoptosis appeared very early in 30 minutes and the number was increased with time pass and there was dose-dependency. Carbon tetrachloride group also showed increased number of apoptotic cells; 0.46 apoptotic cells among 100 liver cell nuclei after 12 hours; 1.85 after 15 hours 1.88 after 24 hours. ApopTag staining showed 10-15% increased number of apoptotic cells than H-E staining in cycloheximide groups, while 2-3 times increased number of apoptotic cells in carbon tetrachloride group. Some of necrotic cells in carbon tetrachloride group were also positive. The results suggested that ApopTag staining is superior than H-E staining with easy detection of the apoptosis, but caution should be paid in interpreting the results because necrotic cell may show positive reaction.

Cycloheximide 가 Mouse 血漿의 Cholinesterase에 미치는 影響

나동진 서울保健大學 1982 論文集 Vol.2 No.1

Cycloheximide의 복강투여가 mouse 혈장 중 colinesterase활성에 미치는 영향을 관찰한 바 투여후 4시간에 암수 공히 현저한 억제효과를 나타내어 50㎍/head(20 g) 투여로 10%, 100 ㎍/head(20g) 투여로 22.5%, 250㎍/head(20g) 투여로 24%의 억제효과를 보였다. The effect of cycloheximide on the activity of cholinesterase were investigated in mouse plasma. Cycloheximide inhibited markedly the activity of cholinesterase in mouse plasma and its inhibition effect was 10% at 50 ㎍/head(20g), 22.5% at 100㎍/head(20g), 24% at 250 ㎍/head(20g). It is suggested from these results that the effect of cycloheximide on the cholinesterase activity is shown by the inhibition of the biosynthesis of this enzyme.

6-Dimethylaminopurine 및 Cycloheximide의 처리가 돼지난포란의 단위발생에 미치는 영향

김종화,박병권,한만희,이규승 충남대학교 농업과학연구소 2007 농업과학연구 Vol.34 No.1

This study is aimed at finding an optimum density for 6-Dimethylaminopurine (6-DMAP) and cycloheximide which have an effect on the revitalization of porcine oocytes. The results were summarized as follows: 1. When 6-DMAP was treated with 2 mM for 2 hours. It showed a significantly (P<0.05) different high result in activation rate, cleavage rate and blastocyst growth rate of 51.2%, 52.7% and 25.2% respectively. 2. When Cycloheximide was treated with 5 ug/ml for 6 hours, It showed a significantly (P <0.05) different high result in activation rate, cleavage rate and blastocyst growth rate of 47.7%, 46.8%, and 27.3% respectively. 3. When it was cultured in the culture medium, NCSU, for 7 days after inducing activation with 6-DMAP and cycloheximide, it showed no differences in the number of inner cell mass (ICM) and total cell of blastocysts. To conclude, it has been examined for porcine oocytes to be suitable when 6-DMAP was treated with 2 mM for 2 hours, Cycloheximide with 5 ug/ml for 6 hours.

항균제 및 항진균제 첨가를 통한 효모 및 유산균의 선택배지 제조

박호민(Homin Park),차정민(Jeongmin Cha),박성은(Seong-Eun Park),손홍석(Hong-Seok Son) 한국식품과학회 2022 한국식품과학회지 Vol.54 No.5

본 연구에서는 항균제인 chloramphenicol과 항진균제인 cycloheximide, amphotericin B, methyl paraben을 각각 YM 배지와 MRS 배지에 첨가하였을 때 배지의 선택성을 확인하였다. Chloramphenicol은 0.01 g/L 이상의 농도에서 유산균의 생육을 억제하였고, cycloheximide는 0.5 g/L 이상의 농도에서 효모의 생육을 억제하는 효과를 나타내었다. 반면 amphotericin B와 methyl paraben은 각각 0.125-0.5, 0.28 g/L의 농도에서는 효모의 생육을 거의 억제하지 못했다. MRS 배지에 지시약인 bromocresol green을 0.05 g/L 첨가했을 때 L. fermentum과 S. cerevisiae 콜로니를 육안상으로 구별하는데 도움이 되었다. 결론적으로 MRS 배지에서 유산균의 생균수 측정을 위해서는 항진균제로 0.5 g/L 이상의 농도의 cycloheximide를, YM 배지에서 효모의 생균수 측정을 위해서는 항균제로 0.01 g/L 이상의 농도의 chloramphenicol을 첨가하는 것이 바람직하다. 실제 막걸리 시료에 chloramphenicol과 cycloheximide를 적용한 결과 효과적으로 배지의 선택성을 증가시킬 수 있음을 확인하였다. 본 연구는 김치와 막걸리와 같이 유산균과 효모가 같이 존재하는 식품을 MRS, YM 배지를 이용해 유산균이나 효모의 생균수만을 확인하고자 할 때 선택배지 제조에 있어 활용이 가능할 것으로 판단된다. 하지만 다양한 미생물이 상이하게 존재하는 여러 발효식품에서도 유효한지에 관해서는 추가적이 연구가 필요하다. In this study, we examined whether the of yeast mold agar and de Man, Rogosa and Sharpe (MRS) agar was improved when antibacterial and antifungal agents were added. The addition of 0.01 g/L of chloramphenicol, a widely used antibacterial agent, was sufficient to inhibit the growth of lactic acid bacteria (LAB), and the addition 0.5 g/L of the antifungal agent, cycloheximide, inhibited yeast growth. The antifungal agents, amphotericin B and methyl paraben, poorly inhibited yeast growth at concentrations of 0.5 and 0.28 g/L, respectively. Moreover, it was shown that the addition of chloramphenicol or cycloheximide selectively inhibited the growth of LAB or yeast in the makgeolli sample, whereby both yeast and LAB were present. These results suggest that these selective media can be used to count in various types of samples in which LAB and yeast coexist.

효율적인 돼지 복제수정란 생산에 관한 연구 1. Ethanol, Ca/sup 2+/ Ionophore, 6-DMAP, Cycloheximide의 농도와 노출시간이 돼지난자의 활성화와 발달에 미치는 영향

위갑인,김광현,강만종,문승주 한국동물생명공학회(구 한국동물번식학회) 2003 Reproductive & developmental biology Vol.27 No.2

활성화를 통한 수핵란의 대량확보를 위해 44시간동안 체외 성숙된 돼지 난자를 ethanol, Ca/sup 2+/-ionophore, 6-DMAP 및 cycloheximide의 화학물질들을 사용하여 단위발생을 유기한 후 그들의 가장적합한 처리농도 및 노출 시간을 규명하였다. 1. Ethanol은 10%, 10분 처리가 전핵형성율, 난할율 및 배발달율에 있어 각각 약 53.4%, 51.6%, 그리고 39.9%로 가장 적합한 조건으로 판명되었다. 2. Ca/sup 2+/-ionophore 가장 적합한 난활성화 조건은 25μM에서 2분간 처리한 것이며, 전핵형성율, 난할율 및 배발달율은 각각 약 59.7%, 62.2%, 그리고 43.9%를 보였다. 3. 6-DMAP를 처리하여 돼지 난자의 활성화를 유기하였을 경우 2mM의 농도에서 각각 약 57.3%, 58.4% 및 29.0%의 전핵형성율, 난할율, 그리고 배발달율을 보여 가장 적합한 조건을 보였으며 2시간∼4.5시간 사이의 노출에는 영향을 받지 않았다. 4. Cycloheximide는 5㎍/ml의 농도가 전핵형성율 2.1%, 난할율 47.7%, 배발달율 31.8%로 가장 은 효율을 보였고, 노출시간에서는 4시간∼6시간 동안 처리하였을 때 60.5∼65.8%, 63.6∼66.7% 및 39.0∼39.5%로 가장 적합한 조건으로 판명되었다. 이상의 결과들은 돼지 체외 성숙 난자의 활성화에 있어 각 화학물질들의 적합한 조건을 바탕으로한 중복처리 및 병용처리 조건 확립 및 효율적인 수핵란의 확보에 기여할 수 있을 것이다. This study was conducted to investigate the optimal condition for produce of large quantity recipient oocytes on porcine cloned embryos. In order to determined the optimum concentration and exposure time of ethanol, Ca/sup 2+/-ionophore, 6-DMAP and cycloheximide, in vitro matured oocytes were activated in TCM-199 containing various chemicals and 15% FBS. The activated oocytes were cultured in Whitten''''s medium containing 10% FBS at 5% CO₂. 1. When the porcine oocytes were activated with the ethanol, the best pronucleus formation, cleavage, and in vitro development rate were obtained in the 10% for 10 minutes, which was significantly higher than all of the other treatment(53.4%, 51.6% and 39.9%, respectively). 2. When the porcine oocytes were activated with the Ca/sup 2+/-ionophore, the pronucleus furmation, cleavage, and in vitro development rate were found significantly higher in the 25μM fur 2min. treatment than those of other concentration and exposure time(59.7%, 62.2% and 43.9%, respectively). 3. When the porcine oocytes were activated with the 6-DMAP, the best pronucleus formation, cleavage, and in vitro development rate were obtained in the 2mM for 2hr∼4.5hr(57.3%, 58.4% and 29.0%, respectively). 4. When the porcine oocytes were activated with the cycloheximide, result showed that pronucleus formation, cleavage, and in vifro development rate were 52.1%, 47.7% and 31.8%, respectively, in the 5㎍/ml for 4hr∼6hr treatmrent, which was significantly higher than all of the other treatment. These results suggested that the active condition of porcine oocytes was established by optimum concentration and exposure time among different chemicals for produce of large quantity recipient oocytes.

Neurospora crassa를 이용한 Tyrosinase 생산의 최적화

채희정,유영제 한국산업미생물학회 1991 한국미생물·생명공학회지 Vol.19 No.3

Neurospora crassa는 성장이 끝나고 분화되는 과정에서 tyrosinase를 생산하며 아미노산류의 기질유사체, 반대사물질, 단백질 생합성 저해제 등에 의해 효소생산이 유도된다. 미생물 균체를 배양시키며 여러 가지 유도물질을 사용하여 효소의 생합성을 유도시킨 결과 가장 적합한 유도물질은 cycloheximide였다. 이는 균체의 성장을 크게 저해하는 항생제의 일종이므로 효소 생합성을 최대화시키기 위한 최적의 농도가 존재하였으며, 유도물질의 농도가 일정한 경우에 효소생산에 최적인 유도시간이 존재하였다. 유도물질에 의한 균체의 성장저해, 기질소모, 유도물질 농도 및 시간에 따른 효소생산 kinetics를 고려하여 수학적 모델링을 하였고 이를 효소생산조건의 최적화에 이용하였다. 유도조건을 최적화함으로써 최대 효소생산성을 크게 향상시킬 수 있었다. Neurospora crassa (KCTC 6079) produces tyrosinase (EC 1.14.18.1) during sexual differentiation under derepressed conditions in the presence of inducers such as amino acid analogues, antimetabolites or protein synthesis inhibitors. The selection of inducer concentration and induction time as well as inducer type are critical for the optimization of the enzyme production. The best inducer was found to be cycloheximide. Since cycloheximide was toxic to the cells, an optimal inducer concentration and an optimal induction time were determined to maximize the enzyme production from batch cultures. Mathematical models for the cell growth and the enzyme production were proposed and used for process optimization. By optimizing the induction conditions, maximum tyrosinase productivity was increased significantly.

Streptanthus tortus 배양 세포에서 당류고갈이 당류 수송계에 미치는 영향

조봉희 한국식물생명공학회 2000 식물생명공학회지 Vol.27 No.1

당류의 수송속도는 당류 고갈동안 증가되었고, 당류 고갈 3일째에 최대 수송속도를 보였다. 당류 고갈로 유도된 당류 수송계는 cycloheximide에 의해서 완전히 방해되었다 선천적으로 식물에서 당류 수송계는 하나이나 당류 고갈동안 유도된 포도당 수송은 포화상태를 보이고, 설탕 수송은 포화되지 않았다. 유도된 당류 수송계는 선천적인 당류 수송계와는 다른 kinetic을 보였다. 이 결과는 고등식물도 불리한 조건하에서 새로운 수송계를 유도시킬 수 있는 능력이 있음을 보여주었다. In suspension cultures of Streptanthus. the uptake rate of sugar was increased during the ceil starvation of sugar in the medium. The maximal uptake rate obtained with 3 days of cell starvation. Sugar transport system induced by the sugar starvation was completely inhibited by 10 $\mu$M cycloheximide. Plant cells are known to possess only one sugar transport system, but the uptake rate of glucose obtained a saturated kinetic while the one of sucrose had two different kinetics after the sugar starvation. Induced sugar transport systems had different kinetics compared to plant cell. These results showed that higher plants have adaptable ability to induce new sugar transport systems when the environment changed unsuitable.