메스암페타민 자발섭취가 흰쥐 뇌조직 중 세로토닌 수용체에 미치는 영향

박기숙(Ki Sook Park),홍진태(Jin Tae Hong),한진이(Jin Yi Han),김혜진(Hye Jin Kim),김용규(Yong Kyu Kim),이종권(Jong Kwon Lee),안광수(Kwang Soo Ahn),이선희(Sun Hee Lee) 한국응용약물학회 2001 Biomolecules & Therapeutics(구 응용약물학회지) Vol.9 No.2

(+)-Methamphetamine (METH) is a psychostimulant, which has been the most popular abused drug in Korea. The rewarding mechanism in METH abuse has been reported to be mediated by dopaminergic system. Recently, it has been reported that dopamine releaser (phentermine) plays a dominant role in the discriminative stimulus effects of METH, whereas 5-HT releaser (fenfluramine) can strongly modify METH selfadministration. The present study is designed to assess the behavioral changes and the changes of the serotonin receptors in the brains of rats administered repeated or self-administered METH. The repeated administration of 1.0 ㎎/㎏/day METH for 12 days increased locomotor activities, and there was no difference between i.v. and i.p. treatment. Rats had actively acquired METH self-administration for 3 weeks at 0.1 or 0.2 mg/kg/injection. Whereas, it was taken few days to acquire sucrose pellet self-administration. The binding of [³H]-8-hydroxy-DPAT (5-HT_(1A) receptors) and [³H]-5-carboxytryptamine (5-HT_(1B) receptors) to brain sections was examined. Both passive administration and self-administration of METH did not change significantly the serotonin receptors levels in hippocampus, striatum and nucleus accumbens. These results suggest that serotonin receptors may not change in the acquisition period of METH self-administration, and we are trying to investigate the serotonin receptors levels of brain in rats maintained of METH self-administration.

소의 간 미크로좀 Aldehyde Dehydrogenase 의 정제 및 특성에 관한 연구

박기숙,주충노 ( Ki Sook Park,Chung No Joo ) 생화학분자생물학회 1994 BMB Reports Vol.27 No.5

Bovine liver microsomal aldehyde dehydrogenase (ALDH) was purified 180 fold by DEAF-Sephacel, Blue Sepharose CL-6B, and 5`-AMP Sepharose affinity column chromatography. The molecules weight of the native enzyme was determined to be 234,000 dalton by Sephacryl S-200 gel filtration. Analysis of the purified enzyme on SDS-polyacrylamide gel electrophoresis showed that it was composed of four identical subunits and the molecular weight of a subunit was 54,900 dalton. Optimum pH of the enzyme was found to be 9.0∼9.5 and it was stable at 25℃ but the activity was decreased as much as 44 percent after incubation at 37℃ for 10 min. The K_m values for short chain aliphatic aldehydes and aromatic aldehyde were 10^(-3) M level but the K_m values for long chain aliphatic aldehydes (C_7∼C_9) were relatively low (10^(-6) M level). These results suggest that the microsomal aldehyde dehydrogenase may play an important role in the oxidation of fatty aldehydes which can be produced during the lipid peroxidation whithin the microsomal membranes. The enzyme showed a sigmoidal saturation curve when nonanal was used as a substrate and the number of the binding sites was determined to be 4 by modified Lineweaver-Burk Plot and Hill Plot. [S]_(0.5) was found to be 18.9 × 10^(-6) M. These results suggest that the enzyme might be stimulated when the concentration of nonanal increases.

소의 간 미크로좀 Aldehyde Dehydrogenase의 정제 및 특성에 관한 연구

박기숙,주충노,Park, Ki-Sook,Joo, Chung-No 생화학분자생물학회 1994 한국생화학회지 Vol.27 No.5

소의 간 미크로좀 분획으로부터 aldehyde dehydrogenase[ALDH, EC 1.2.1.3]를 sodium deoxycholate 처리, DEAE-Sephacel, Blue Sepharose CL-6B, 그리고 5'-AMP Sepharose 4B affinity 등의 column chromatography 방법으로 186배 정제하였다. Sephacryl S-200 gel filtration 방법으로 측정된 이 효소의 분자량은 293.9KDa이었고 SDS-polyacrylamide gel 전기영동법을 수행한 결과 subunit 분자량이 54.9KDa인 homotetramer임이 판명되었다. 최적 pH 범위는 9.0~9.5로 비교적 넓었으며 $25^{\circ}C$에서는 안정하지만 $37^{\circ}C$에서 10분간 방치하면 효소활성이 44% 감소되었다. 탄소수가 짧은 aliphatic aldehydes나 aromatic aldehyde에 대한 $K_m$값은 $10^{-3}$ M 수준이었으나 탄소수가 긴 aliphatic aldehyde($C_7{\sim}C_9$)에 대한 $K_m$값은 $10^{-6}$ M 수준으로 아주 낮은 것으로 보아 미크로좀 ALDH가 미크로좀 막에서 lipid peroxidation과정에서 생성되는 fatty aldehydes의 산화에 중요한 역할을 할 것으로 생각된다. 또한 이 효소는 nonanal을 기질로 사용했을 경우에만 속도-기질농도 곡선이 sigmoidal saturation curve를 보였고 modified Lineweaver-Burk Plot과 Hill Plot을 행한 결과 결합자리수가 4임이 밝혀졌다. Nonanal의 $[S]_{0.5}$는 $18.9{\times}10^{-6}M$이었고 nonanal의 농도가 증가하면 효소활성이 크게 증가됨을 알 수 있었다. 이 효소의 활성 부위의 아미노산 잔기를 조사하기 위하여 sulfhydryl기에 특이하게 반응하는 NEM, DNTB를 처리했을 때는 효소의 활성이 부분적으로 억제되었으나 hydroxyl기에 특이하게 반응하는 PMSF를 처리했을 경우 효소활성의 억제 정도가 매우 낮은 것으로 보아 효소활성 부위에 sulfhydryl기가 존재할 것으로 생각된다. 또한 ALDH에 대한 inhibitor로 알려진 disulfiram에 대해서도 매우 민감하게 저해되지만 0.2mM의 높은 disulfiram 농도에서도 26%의 효소활성이 남아 있는 것으로 보아 disulfiram 작용 부위와 효소활성 부위는 동일하지 않은 것으로 생각된다. Bovine liver microsomal aldehyde dehydrogenase (ALDH) was purified 180 fold by DEAE-Sephacel, Blue Sepharose CL-6B, and 5'-AMP Sepharose affinity column chromatography. The moleculer weight of the native enzyme was determined to be 234,000 dalton by Sephacryl S-200 gel filtration. Analysis of the purified enzyme on SDS-polyacrylamide gel electrophoresis showed that it was composed of four identical subunits and the molecular weight of a subunit was 54,900 dalton. Optimum pH of the enzyme was found to be 9.0~9.5 and it was stable at $25^{\circ}C$ but the activity was decreased as much as 44 percent after incubation at $37^{\circ}C$ for 10 min. The $K_m$ values for short chain aliphatic aldehydes and aromatic aldehyde were $10^{-3}$ M level but the $K_m$ values for long chain aliphatic aldehydes ($C_7{\sim}C_9$) were relatively low ($10^{-6}$M level). These results suggest that the microsomal aldehyde dehydrogenase may play an important role in the oxidation of fatty aldehydes which can be produced during the lipid peroxidation whithin the microsomal membranes. The enzyme showed a sigmoidal saturation curve when nonanal was used as a substrate and the number of the binding sites was determined to be 4 by modified Lineweaver-Burk Plot and Hill Plot. $[S]_{0.5}$ was found to be $18.9{\times}10^{-6}M$. These results suggest that the enzyme might be stimulated when the concentration of nonanal increases.

안전성약리시험의 Good Laboratory Practice 평가기술연구

최기환(Ki Hwan Choi),박기숙(Ki Sook Park),이윤희(Yun Hee Lee),나한광(Hang-Kwang Na),윤재석(Jae Suk Yun),김동섭(Dong-Sup Kim),김주일(Joo-Il Kim) 한국독성학회 2006 Toxicological Research Vol.22 No.2

Safety pharmacology studies are conducted to investigated the potential undesirable pharmacodynamic effects of a substance on physiological functions in relation to exposure in the therapeutic range and above. In the International Conference on Harmonisation (ICH), the guideline “S7A : Safety Pharmacology Studies for Human Pharmaceuticals” has been developed and reached Step 5 of the ICH process in 2001. Now the Korea Food and Drug Administration (KFDA) are going to transfer “The Guideline for General Pharmacology” into “The Guideline for Safety Pharmacology”. Safety pharmacology studies should be performed in compliance with Good Laboratory Practice (GLP). Thus, the present paper reviews the Japanese GLP guidelines for pharmaceuticals to help the conduct and inspection of safety pharmacology studies in compliance with GLP. We also reviewed the ICH guidelines “S7B revised : The Nonclinical Evaluation of the Potential for Delayed Ventricular Repolarization (QT Interval Prolongation) by Human Pharmaceuticals” and “E14 : The Clinical Evaluation of QT/QTc Interval Prolongation and Proarrhythmic Potential for Non-antiarrhythmic Drugs” to apply our drug approval systems.

간경변이 동반된 임신 3례

이기주(Ki Joo Lee),문정빈(Jeong Bin Moon),한수연(Soo Yeon Han),김미하(Mi Ha Kim),기숙현(Sook Hyeun Kee),박중신(Joong Shin Park),전종관(Jong Kwan Jun),윤보현(Bo Hyun Yoon),신희철(Hee Chul Syn) 대한산부인과학회 1999 Obstetrics & Gynecology Science Vol.42 No.9

Far Eastern countries including Korea show the high prevalence of hepatitis B virus carriers, so that the incidence of liver cirrhosis is higher than in western countries. But pregnancies with liver cirrhosis are rarely encountered in clinical settings, since liver cirrhosis usually develops after childbearing ages and often causes the disturbance of estrogen metabolism, resulting in severe menstrual irregularity and infertility. Therefore, little is known about the interactions between liver cirrhosis and pregnancy. Liver cirrhosis and portal hypertension are not contraindications to pregnancy but necessitate intensive monitoring throughout pregnancy because the complications of liver cirrhosis, which pose additional risks during pregnancy, are numerous and unpredictable. We report 3 cases of pregnancies in patients with liver cirrhosis with brief review of the literature.

G009 의 간 보호작용에 관한 연구

고광호(Kwang Ho Ko),박기숙(Ki Sook Park),이주영(Joo Young Lee),정진호(Jin Ho Chung),조미정(Mee Jung Cho),이준우(Jun Woo Lee),정훈(Hoon Jeong),이승룡(Seung Yong Lee) 한국응용약물학회 1994 Biomolecules & Therapeutics(구 응용약물학회지) Vol.2 No.2

The present study was performed to determine the protective effect of G009 on liver damage induced by ethanol, CCl₄, and thioacetamide in rats. In acute fatty liver animal model induced by ethanol, triglyceride accumulation was markedly decreased to the normal control level by 25 ㎎/㎏ G009 treatment. In addition, G009 significantly reduced serum ALT and AST levels in CCl₄-induced acute hepatitis animals. Treatment of G009 to the acute hepatitis rats induced by thioacetamide resulted in a dose dependent reduction of serum ALT level as well as AST level up to the normal control level. These protective effects of G009 were confirmed by histological examinations of the liver. These results suggested that G009 could be effective for the protection from the liver damage induced by ethanol, CCl₄ and thioacetamide.

흰쥐 배양 전배자 및 중뇌세포에서 Ochratoxin A의 독성

홍진태(Jin Tae Hong),박귀례(Kui Lea Park),한순영(Soon Young Han),박기숙(Ki Sook Park),김형식(Hyung Sik Kim),오세동(Se Dong Oh),박희정(Hee Jung Park),이이다(Rhee Da Lee),장성재(Seung Jae Jang) 대한약학회 1998 약학회지 Vol.42 No.3

Effects of ochratoxin A (OTA) on embryo development were studied in cultured whole embryos from 9.5 day gestation rat for 48 h. OTA (more than 0.5mcg/ml) induced microcephaly in the cultured rat whole embryos. Protein and DNA content, and DNA synthesis were significantly inhibited by OTA. We next examined whether the microcephaly seen in cultured whole embryo partially results from inhibition of differentiation of embryonic midbrain cells. Embryonic midbrain cells were extracted from 12 day gestation rat embryos, and cultured for 96 hr. OTA ibhibited cell differentiation about 50% over control. We also tested whether OTA-induced embryotoxicity would be associated with oxidative damages. We measured the gamma-glutamyltranspeptidase (gamma-GT) and glutathione peroxidase (GPX) activities, and glutathione (GSH) content in both cultured whole embryos and embryonic midbrain cells. OTA decreased GSH content, whereas slightly increased gamma-GT activity, but GPX activity was not significantly changed. These results show that OTA caused the microcephaly and its effect may be partially due to the inhibition of cell differentiation of embryonic midbrain cells, but the role of oxidative damages is not clear in embryotoxicity.

화장품 원료의 피부자극성과 세포독성의 관련성

이은희(Eun Hee Lee),이종권(Jong Kwon Lee),김용규(Yong Kyu Kim),박기숙(Ki Sook Park),안광수(Kwang Soo Ahn),정경미(Kyoung Mi Jung),정해관(Jung Hai Kwan),이선희(Sun Hee Lee),정수연(Soo Youn Chung),홍진태(Jin Tae Hong) 대한약학회 2001 약학회지 Vol.45 No.3

To compare skin irritation and cytotoxicity of anti-wrinkle agents, we examined skin irritation of six anti-wrinkle agents (ascorbic acid, glycolic acid, all trans-retinoic acid, ginseng extract, retinol, EB) in New Zeland white rabbit. Cytotoxicity of these agents was determined by MTT [tetrasolium salt 3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyl tetrazolium bromide] at multi-time points in cultured HaCaT cell, a human immortalized keratinocyte cell. We then analyzed correlation between skin irritation and cytotoxicity by spearman's rank correlation analysis. All trans- retinoic acid shewed the highest primary irritation index (0.92) in skin irritation test. Being all the six agents not irritant, retinol showed the most cytotoxic agents. The correlation between skin irritation and cytotoxicity IC50 at different time point was 0.814,0.757,0.814 and 0.7 at 3,24, 48 and 72h, respectively. We also found that IC20 and IC80 of these agents showed similar correlation with skin irritation. These results therefore demonstrated that there is close correlation between skin irritation and cytotoxicity IC50 value by MTT in HaCaT cell at early time points by anti-wrinkle agents or IC20value. IC50 at earily time point or IC20 values may be reliable alter- native determinant of skin irritation.

Phthalate의 피부자극시험 및 안점막자극시험에 관한 연구

이종권(Jong Kwon Lee),김주환(Ju Hwan Kim),이은희(Eun Hee Lee),김용규(Yong Kyu Kim),홍진태(Jin Tae Hong),박기숙(Ki Sook Park),안광수(Kwang Soo Ahn),정수연(Soo Youn Chung),이선희(Sun Hee Lee) 한국독성학회 2001 Toxicological Research Vol.17 No.2

Phthalates are widely used as plasticizers to impart softness and flexibility to normally rigid polyvinylchloride products. However, there are not much studies for dermal and ocular irritation toxicity of phthalates. So we investigated the skin or eye irritation effect of some phthalates which was not reported. The primary skin irritation of diethyl phthalate (DEP), diisodecyl phthalate (DIDP) , diisononyl phthalate (DINP), dipropyl phthalate (DPP) and dipropyl phthalate (DPrP) was studied. The ocular irritation of dibutyl phthalate(DBP), DIDP, DINP, DPP and DPrP was also studied. DEP, DIDP, DINP, DPR, and DPrP were found to be non-irritating to the skin of the test animals. DBP, DIDP, DINP and DPP were found to be non-irritating to the eye of the rabbits. DPrP caused the slight irritations to the eye in 1 or 2 days after treatment but irritation of the animals was soon recovered.

일산화질소에 의해 유도된 연골 세포 사멸에서의 항세포사멸유전자 Bcl-XL의 방어 효과

한창환 ( Chang Whan Han ),신윤학 ( Yun Hak Shin ),권순용 ( Soon Yong Kwon ),조윤경 ( Yun Kyoung Cho ),지보근 ( Bo Keun Jee ),김영율 ( Young Yul Kim ),김순희 ( Soon Hee Kim ),박기숙 ( Ki Sook Park ),강길선 ( Gil Son Khang ) 한국조직공학과 재생의학회 2005 조직공학과 재생의학 Vol.2 No.2

To evaluate whether transfection of human chondrocytes with an anti-apoptotic gene, Bcl-XL, can prevent nitric oxide (NO)-induced chondrocyte apoptosis. We stably transduced early passage chondrocytes with an anti- apoptotic gene Bcl-XL and retrovirus. Bcl-XL transducants were compared with chondrocytes transduced in parallel with a Lac-Z gene. After the cells were treated with 500 microM of SNP, the process of apoptosis was assessed by trypan blue exclusion, enzyme-linked immunosorbent assay (ELISA), and flow cytometry. The functional aspect of the chondrocyte was measured by proteoglycan synthesis assay. Chondrocytes were cultivated in NO, the chondrocytes that were either not transfected or transfected with Lac-Z exhibited a decrease in viable cell number in comparison to the Bcl-XL transfected chondrocytes. The NO significantly increased fragmentation of DNA in chondrocytes that were either not transfected or transfected with Lac-Z, and this increase in DNA fragmentation was significantly greater than in the chondrocytes transfected with Bcl-XL. Flow cytometric result showed that apoptotic chondrocytes were significantly higher in the transfected and Lac-Z transfected chondrocytes than Bcl-XL transfected chondrocytes. Chondrocytes transfected with Bcl-XL were protected from NO-induced impairment of proteoglycan synthesis response to NO. From the present study, we have demonstrate that NO-induced chondrocytes death involve mechanisms subjected to regulation by an anti-apoptotic protein, Bcl-XL. Therefore, Bcl-XL gene therapy has the potential to protect apoptosis in human chondrocyte.