흰 쥐 간으로부터 분리된 Microsomal Cytochrome P - 450 dependent Monooxygenase 의 안정성

김숙영,박기현 ( Sook Young Kim,Ki Hyun Park ) 생화학분자생물학회 1988 BMB Reports Vol.21 No.4

The stability of cytochrome P-450, cytochrome b_5, cytochrome P-450-dependent monooxygenases, and NADPH-cytochrome P-450 reductase was observed in liver microsomes prepared from rats pretreated with β-naphthoflavone during the stored period of 8 months at -80℃. The contents of P-450 and b_5 were not changed significantly and the activities of aryl hydrocarbon hydroxylase, 7-ethoxycoumarin ο-deethylase, and benzphetamine demethylase were also stable for this period. However, the activities of NADPH-cytochrome P-450 reductase were increased gradually, whereas since 5th month, those of testosterone hydroxylases including 16α-hydroxylase were decreased. The decrease seems due to a certain change in testosterone binding site of P-450, not to the inactivation of one isozyme of P-450.

Saponin을 함유한 생약추출물의 거담작용에 관한 연구

김숙영(Sook-Young Kim),문자영(Ja-Yeong Moon),이동욱(Dong-Wook Lee),박기현(Ki-Hyun Park) 한국생약학회 1988 생약학회지 Vol.19 No.2

The effect of ethanol extracts of Gilkyung(Platicodi Radix, Platicodon grandiflorum A, DC), Onji(Polygalae Radix, Polygala tennuifolia Willdenow) and Deoduk(Codonopsis lanceolate Radix, Codonopsis lanceolata) on expectorant activity of rat trachea was investigated. Following treatment of 50% ethanol extract of these medicinal plants (25 ㎎/rat), the content of neurotransmitters such as acetylcholine and histamine in tissue was significantly increased. The secretions of acid glycoproteins and the artificially injected phenol red were also increased. However, there was no significant difference except Onji From the histological study through periodic acid Schiff and alcian blue stain, the thickness of inner membrane of acinar glands and the stained glycoproteins on surface of epithelium and on the glands were observed in all the rats trachea treated with extract of medicinal plants. In vitro, the viscosity of mucin solution was slightly decreased with an addition of the extracts. Onji showed the most effective expectorant activity among them at the identical conditions. The mechanism of expectorant activity of these medicinal plants seems to be due to stimulation of secretion and changes of rheological properties of mucus.

흰 쥐 Strain에 따른 간 마이크로좀의 Cytochrome P-450 동위 효소 특성

김숙영,박기현,Kim, Sook-Young,Park, Ki-Hyun 생화학분자생물학회 1986 한국생화학회지 Vol.19 No.3

Cytochrome P-450 (P-450) 동위 효소 유도 물질로서 phenobarbital (PB) 또는 3-methylcholanthrene (MC)을 처리한 흰 쥐 strain (Sprague-Dawley, Fischer, Wistar)의 간 마이크로좀에서 총 P-450 함량, aryl hydrocarbon hydroxylase 활성도의 유도 정도 및 P-450 동위 효소의 구성비 등은 strain에 따라 다소 차이를 보였으나, P-450-b type은 PB에 의해, c type은 MC에 의해 주로 유도되었으며, 이들 동위 효소의 기질 특이성, 단일 항체와의 결합에 의한 효소 활성도의 억제, 동위 효소의 항체 결합 부위 등은 strain의 종류와 상관관계가 없는 것으로 보였으며 P-450 동위 효소의 이러한 몇가지 특성들이 strain 사이에서 유전적으로 보존되고 있는 것으로 추정되었다. After i.p. administration of phenobarbital (PB) or 3-methylcholanthrene (MC) on the adult male rat strains, Wistar, Fischer, and Sprague-Dawley, we compared the properties of hepatic microsomal cytochrome P-450 (P-450) isozymes. Total P-450 contents, degree of induction of aryl hydrocarbon hydroxylase (AHH) activity and isozyme pattern of P-450 were slighly dependent on rat strains. The major form of P-450 isozymes in rats tested was P-450-b in the case of PB-microsomes and that of MC-microsomes was P-450-c. Substrate specificity of the isozymes and the inhibition of activity by the binding of P-450-specific monoclonal antibodies to the isozymes were not associated with rat strains. The results obtained from immunological assay using monoclonal antibodies indicated that the sensitivity of hepatic P-450-dependent AHH activity was rather influenced by the treatment of PB or MC as inducers than by the differences of strain. Thus, it seemed that such characteristics of P-450 isozymes have been genetically conserved among rat strains.

일차 배양 휜쥐 간세포에서 CCl₄ 유발 세포독성을 이용한 간보호 효과 검색방법

김영숙(Young Sook Kim),박기현(Ki Hyun Park) 한국생약학회 1995 생약학회지 Vol.26 No.1

To devise an in vitro screening method for antihepatotoxic activity, CCl₄-induced cytotoxicities in primary cultures rat hepatocytes were examined. When rat hepatocytes were intoxicated with 0.5, 1.0 or 1.5 mM CCl4 for 1.5, 3 or 19hr, in order of LDH>GOT>GPT release form hepatocytes was increased in a dose-dependent manner. Treatment with 1.5 mM CCl₄ for 1.5 hr showed maximum increase in activity of LDH, GOT or GPT released in the medium compared with the control. At this experimental condition, well known antihepatotoxic substances, glycyrrhizin and silybin markedly inhibited CCl₄-induced cytotoxicities. These results demonstrated that the screening method using CCl₄-induced injury in primary cultured rat hepatocytes might be suitable in vitro assay for antihepatotoxic activity.

수종의 전통약제가 일차 배양 간세포에서 CCl₄ 유발 세포독성에 미치는 영향

김영숙(Young Sook Kim),박기현(Ki Hyun Park) 한국생약학회 1995 생약학회지 Vol.25 No.4

80% Methanol extracts of 44 traditional drugs used for the treatment of liver diseases or tonic effects were screened for anti-hepatotoxic activity by in vitro assay using CCl₄ induced cytotoxicity in primary cultured rat hepatocytes. CCl₄-induced cytotoxicity was evaluated by determination of LDH, GOT or GPT activity in the medium. Rehmaniae Radix Preparata and Gelantina nigra inhibited the release of LDH, GOT and GPT from CCl₄ treated hepatocytes. Gibotii Rhizoma and Eucommiae Cortex showed inhibitory effect on release of LDH from normal hepatocytes as well as CCl₄-treated hepatocytes. Eucommiae Cortex and Lili Bulbus decreased release of GOT and LDH from normal hepatocytes, respectively. Astragali Radix inhibited release of GPT in CCl₄-treated hepatocytes. Phlomidis Radix, Imperatae Rhizoma, Cistanchis Herba, Broussonetiae Fructus, Asparagi Tuber, Trigonellae Semen and Polygonati Rhizoma inhibited release of LDH from CCl₄-treated hepatocytes. Among 44 traditional drugs, most of them released LDH, GOT or GPT at the dose of 1 mg/ml in normal hepatocytes, and Dynariae Rhizoma, Acanthopanacis Cortex, Longanae Arillus, Atratylodis Rhizoma and Ecliptae Herba increased CCl₄-induced cytotoxicity.

일차 배양 흰쥐 간세포에서 사염화탄소 유발 세포독성에 대한 수종 생약 용매 분획의 억제효과 검색과 in vivo 간보호 작용 평가

김영숙(Young Sook Kim),경종수(Jong Su Kyung),박기현(Ki Hyun Park) 대한약학회 1996 약학회지 Vol.40 No.1

Solvent fractions were prepared from traditional herbal drugs which of methanol extracts inhibited CCl4-induced cytotoxicity in primary cultured rat hepatocytes and continuously assayed their effects. Ethylacetate and n-buthanol fractions from Cibotii Rhizoma and chloroform fraction from Gelatina Nigra inhibited the release of LDH and GPT from CCl4-treated hepatocytes, respectively. Water fraction (WAR) among solvent fractions from Astragali Radix showed the most potent inhibitory effect on the release of GOT or GPT by treatment with CCl4. All of solvent fractions prepared from Eucommiae Cortex had no effect on CCl4-induced cytotoxicity. Chloroform and ethylacetate fractions from Rehmanniae Radix Preparata increased the release of GPT from CCl4-treated hepatocytes. n-Hexan, chloroform or ethylacetate fraction from 5 herbal drugs increased the release of LDH, GOT or GPT from normal hepatocytes at the dose of 1.Omg/ml. Administration of WAR suppressed the elevation of GOT, ALP activities and MDA contents in the serum as well as in the liver tissue of CCl4-intoxicated rats. Based on these results, isolation of antihepatotoxic substances from WAR is under the process.

The Stability of Microsomal Cytochrome P-450-dependent Monooxygenases from Rat Liver during Storage in Freezing

김숙영,박기현,Kim, Sook-Young,Park, Ki-Hyun 생화학분자생물학회 1988 한국생화학회지 Vol.21 No.4

이물질 대사에 관여하는 cytochrome P-450(P-450) 의존성 효소들은 동물 조직에서 microsome을 분리한 후 냉장고에 장기 보존하면서 연구 목적에 따라 microsomal monooxygenase 활성도를 관찰하는 경우가 적지 않다. 따라서 ${\beta}$-naphthoflavone을 처리한 흰 쥐의 간 microsome을 $-80^{\circ}C$에서 보존하면서 P-450, cytochrome $b_5(b_5$), 몇 종류의 cytochrome P-450-dependent monooxygenase와 NADPH-cytochrome P-450 reductase 둥의 안정성을 검토하였다. P-450과 $b_5$의 함량은 변화를 보이지 않았으며 aryl hydrocarbone hydroxylase, 7-ethoxycoumarin O-deethylase와 benzphetamine demethylase의 활성도 역시 변화가 없었으나 NADPH-cytochrome P-450 reductase의 활성도는 저장 4개월 이후 점차 증가하였다. 한편 testosterone $16{\alpha}$-hydroxylase를 포함한 testosterone의 hydroxylase들은 저장 5개월 후부터 점차로 감소하였는데 이는 P-450의 testosterone 결합분위의 어떤 변화 때문인 것으로 생각된다. The stability of cytochrome P-450, cytochrome $b_5$, cytochrome P-450-dependent monooxygenases, and NADPH-cytochrome P-450 reductase was observed in liver microsomes prepared from rats pretreated with ${\beta}$-naphthoflavone during the stored period of 8 months at $-80^{\circ}C$ The contents of P-450 and $b_5$ were not changed significantly and the activities of aryl hydrocarbon hydroxylase, 7-ethoxycoumarin O-deethylase, and benzphetamine demethylase were also stable for this period. However, the activities of NADPH-cytochrome P-450 reductase were increased gradually, whereas since 5th month, those of testosterone hydroxylases including $16{\alpha}$-hydroxylase were decreased. The decrease seems due to a certain change in testosterone binding site of P-450, not to the inactivation of one isozyme of P-450.

Flavone 투여로 흰쥐간에서 유도된 Microsomal Cytochrome P - 450 의 특성

김숙영,이동욱,박기현 ( Sook Young Kim,Dong Wook Lee,Ki Hyun Park ) 생화학분자생물학회 1990 BMB Reports Vol.23 No.4

Cytochrome P-450 (P-450) isozymes of flavone-pretreated rats were compared with those of partially purified microsomal proteins and liver microsomes from control rats and phenobarbital (PB)-, 3-methylcholanthrene(MC)-, and isosafrole(ISF)-pretreated rats. The activities of microsomal P-450 dependent aryl hydrocarbon hydroxylase(AHH) and ethoxycoumarin deethylase(ECDE) were more than three times of those of control. AHH and ECDE activities of partially purified P-450 with DE_(52) column chromatography were increased more than eight times of control. Monoclonal MC 1-7-1 antibody, which is specific to P-450 c and d isozyme, bound specifically to liver microsomal P-450 of flavone-rats in radioimmunoassay. Some properties of P-450 of flavone-rats were identical with those of P-450 d in spectral and electrophoretic studies. The absorption maximum of P-450-CO complex of flavone-rats was 448 nm (same as that of MC-pretreated rats), and molecular weight of partially purified P-450 from the major band of electrophoresis was 52,000 (same as that of PB-pretreated rats). Thus, it can be indicated that the pretreatment of flavone induces P-450 d or P-450 isozyme with similar immunochemical properties to P-450 d.

황기(黃耆)의 간기능 보호 성분

김영숙,경종수,박기현,백남인,Kim, Young-Sook,Kyung, Jong-Soo,Park, Ki-Hyun,Baek, Nam-In 한국생약학회 1996 생약학회지 Vol.27 No.2

The components were isolated from the root of Astragalus membranaceus and their structures were characterized as 3,4-methylenedioxypyrrolealdehyde, $7-O-{\beta}-D-glucopyranosyl$ 7, 3'-dihydroxy-4'-methoxy isoflavone and a naphthalene derivative on the basis of spectral and physical methods. $7-O-{\beta}-D-glucopyranosyl$ 7, 3'-dihydroxy-4'-methoxy isoflavone and the naphthalene compound showed protective effect on $CC_{l4}-induced$ cytotoxicity in primary cultured rat hepatocytes.

조기난소부전 환자에서 X 염색체 불활성화 분석

손인숙 ( In Sook Sohn ),유달영 ( Dal Yeong Yoo ),장동욱 ( Dong Wook Jang ),차윤정 ( Yun Jeong Cha ),김수녕 ( Soo Nyung Kim ),이지영 ( Ji Young Lee ),윤병일 ( Byung Il Yun ),정소정 ( So Chung Chung ),박기현 ( Ki Hyun Park ),이병석 대한산부인과학회 2004 Obstetrics & Gynecology Science Vol.47 No.8

목적: 조기난소부전은 난소발달기전을 이해하는 임상적 모델로, 조기난소부전의 유전적 원인을 밝힘으로써 난소분화 및 난포 형성과정에 관련된 유전자를 알 수 있다. 조기난소부전은 40세 이전에 무월경과 성선자극호르몬이 증가되는 임상증후군으로 빈도는 약 1% 정도이다. 아직 조기난소부전의 기전은 밝혀지지 않았고, 터너 증후군, 자가면역질환, 항암제 치료, 방사선 치료 및 감염 등이 원인으로 알려져 있다. 많은 유전자가 조기난소부전의 후보 유전자로 제기되었고, Objective: Premature ovarian failure (POF) is a highly heterogenous condition, and its etiology remains unknown in approximately two-thirds of cases. POF can be caused by Turner syndrome, genetic disease, iatrogenic agents such as chemotherapy and radioth