흰쥐 간 마이크로좀의 cytochrome P - 450 의존성 testosterone hydroxylase 의 활성도 측정

이동욱,박기현 ( Dong Wook Lee,Ki Hyun Park ) 생화학분자생물학회 1988 BMB Reports Vol.21 No.3

A rapid and one time determinable TLC method for the monohydroxytestosterones formed by microsomal cytochrome P-450(P-450) was established, and applying this method the activity of P-450-dependent testosterone hydroxylases was observed in liver microsomes of rats pretreated with phenobarbital(PB). The 14 testosterone metabolites were found on the TLC plate. The amounts of 6-monohydroxytestosterones (2α-, 2β-, 6α-, 6β-,7α-and 16α-OH-T) were determined. The formation of these metabolites showed a good linearity(r = 0.9687-0.9999) corresponding to the concentration of P-450 added up to about 1.0 nmol/min/reaction mixture, and an excellent reproducibility (C.V.= 1.8 ± 1.0%) when determined with 0.5 nmol P-450 in this assay system. The hydroxylation of testosterone to its 2β-OH-T by PB-inducible P-450 was increased (2 fold), while 2α- and 7α-OH-T was rather decreased to 48 and 32%, respectively, compared with those of uninduced one. The 6β-OH-T was predominantly formed by uninducible and PB-inducible P-450, especially PB-inducible P-450 hydroxylated stereospecifically onto the C2-position of testosterone (2α/2β =1: 3.8). However, the oxidation rates of testosterone to its 6α-, 6β- and 16α-OH-T, and the total amount of testosterone metabolites identified were not changed, significantly by the administration of PB.

Assay of Cytochrome P-450-dependent Testosterone Hydroxylases in Liver Microsomes of Rats

이동욱,박기현,Lee, Dong-Wook,Park, Ki-Hyun 생화학분자생물학회 1988 한국생화학회지 Vol.21 No.3

흰쥐 간 마이크로좀의 cytochrome P-450(P-450)에 의해 생성된 testosterone의 monohydroxy 대사물을 동시에 쉽게 정량할 수 있는 TLC 방법을 정립하였다. 이 방법을 이용하여 phenobarbital(PB)로 유도된 흰쥐 간 마이크로좀의 P-450에 의한 testosterone의 대사를 관찰하였다. 생성된 14종의 대사물이 확인되었고 그중 6종의 monohydroxy 대사물($2{\alpha}-$, $2{\beta}-$, $6{\alpha}-$, $6{\beta}-$, $7{\alpha}-$ 그리고 $16{\alpha}$-monohydroxytestosterone)을 정량하였다. 설정한 반응조건에서 이들 대사물의 생성은 P-450의 농도(1nmol/0.5ml 반응액 이하)에 따라 양호한 직선 관계 (r=0.9687-0.9999)와 재현성(C.V=$1.8{\pm}1.0%$)을 나타내었다. 또한 PB로 유도된 P-450에 의한 $2{\beta}$-monohydroxytestosterone의 생성은 대조군에 비해 2배로 증가된 반면$2{\alpha}-$와 $7{\alpha}$-monohydroxytestosterone의 생성은 오히려 48, 32%로 각각 감소되었다. 그러나 확인된 대사물의 총함량은 서로 비슷하였다. A rapid and one time determinable TLC method for the monohydroxytestosterones formed by microsomal cytochrome P-450(P-450) was established, and applying this method the activity of P-450-dependent testosterone hydroxylases was observed in liver microsomes of rats pretreated with phenobarbital(PB). The 14 testosterone metabolites were found on the TLC plate. The amounts of 6-monohydroxytestosterones ($2{\alpha}-$, $2{\beta}-$, $6{\alpha}-$, $6{\beta}$, $-7{\alpha}$-and $16{\alpha}$-OH-T) were determined. The formation of these metabolites showed a good linearity(r = 0.9687-0.9999) corresponding to the concentration of P-450 added up to about 1.0 nmol/min/reaction mixture, and an excellent reproducibility (C.V.=$1.8{\pm}1.0%$) when determined with 0.5 nmol P-450 in this assay system. The hydroxylation of testosterone to its $2{\beta}$-OH-T by PB-inducible P-450 was increased (2 fold), while $2{\alpha}-$ and $7{\alpha}$-OH-T was rather decreased to 48 and 32%, respectively, compared with those of uninduced one. The $6{\beta}$-OH-T was predominantly formed by uninducible and PB-inducible P-450, especially PB-inducible P-450 hydroxylated stereospecifically onto the C2-position of testosterone $(2{\alpha}/2{\beta}=1:3.8)$. However, the oxidation rates of testosterone to its $6{\alpha}-$, $6{\beta}-$ and $16{\alpha}$-OH-T, and the total amount of testosterone metabolites identified were not changed significantly by the administration of PB.

Diethylnitrosamine 과 Diethanolnitrosamine 이 흰쥐 혈청의 탈수소 효소활성에 미치는 영향

장성근,이동욱,박기현 ( Sung Keun Chang,Dong Wook Lee,Ki Hyun Park ) 생화학분자생물학회 1991 BMB Reports Vol.24 No.6

Lactate dehydrogenase(LDH) and its isozyme patterns were investigated in the serum of the rats treated with diethylnitrosamine(DENA) or diethanolnitrosamine(DELNA). LDH activity was remarkably increased by the treatment of these chemicals, but that was gradually decreased from about 2 weeks after administration of the chemicals. Especially the relative activity of LDH isozyme 1 was increased remarkably, whereas that of isozyme. 5 was slightly decreased from the 5th days. However, the activity ratios of the isozymes were not changed significantly with time course. This unique alterations in serum LDH activity might be provide a useful information for the detection of tissue toxicity induced by these chemicals.

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.

성선 이형성 환자 혈액 및 성선 조직에서 Y 염색체 모자이시즘의 진단

김진영,이상준,박기현,김정연,배상욱,이병석,김세광,김인규,조동제,송찬호,김재욱,이호준,Kim, Jin-Yeong,Lee, Sang-Joon,Park, Ki-Hyun,Kim, Jung-Yeon,Bai, Sang-Wook,Lee, Byung-Seok,Kim, Se-Kwang,Kim, In-Kyu,Cho, Dong-Je,Song, Chan-Ho,Kim, Jae- 대한생식의학회 1999 Clinical and Experimental Reproductive Medicine Vol.26 No.3

Objective: The presence of Y chromosome in patients with gonadal dysgenesis is related to the risk of gonadoblastoma. Since the patients with abnormal sexual differentiation may have cryptic Y mosaicism, it is important to detect the presence of Y material in these patients. But sometimes it is difficult to detect Y material only with karyotyping. This study was performed to evaluate the usefulness of the SRY gene screening in blood and gonad by using PCR in detecting the presence of Y material and possible tissue mosaicism in patients with gonadal dysgenesis as Turner syndrome and 46,XY pure gonadal dysgenesis (PGD, Swyer syndrome). Method: In 26 patients with gonadal dysgenesis, we screened for Y material by using PCR for SRY gene in peripheral leukocytes and in gonadal tissues of some patients. They were 22 cases of Turner syndrome (7 45,XO, 2 46,Xi(Xq), 3 45,XO/46,XX, 5 45,XO/46,Xi(Xq), 1 45, XO/46,XY, 1 45,XO/46,Xi(Yq), 1 45,XO/47,XYY, 1 46,XX,del(X)(q24) and 1 46,X,+mar) and 4 cases of 46,XY pure gonadal dysgenesis. PCR for SRY gene in the gonadal tissue was performed in 5 Turner syndrome and 2 PGD to determine the cryptic Y mosaicism between blood and gonad. Results: By using PCR analysis for SRY, Y chromosome material was detected in the blood of 4 of 22 Turner syndrome patients (45,XO/46,Xi(Xq), 45,XO/46,Xi(Yq), 45,XO/46,XY, and 45, XO/47,XYY), 3 of 4 46,XY pure gonadal dysgenesis. Discrepancy between karyotyping and blood PCR for SRY was noted in 1 Turner syndrome (45,XO/46,Xi(Xq)) and 1 PGD. Laparoscopic gonadectomy was performed in Y containing or SRY positive cases. In addition, PCR analysis for SRY in the gonads of 5 Turner syndrome and 2 PGD showed discrepancy between blood and gonad or between both gonads in 3 Turner syndrome (45,XO/46,Xi(Xq), 45,XO/46,Xi(Y q), 45,XO/46,XY) and 2 PGD patients. Conclusion: In gonadal dysgenesis, PCR analysis for SRY gene is useful to detect the cryptic Y mosaicism that is sometimes undetected by karyotyping. And since there may be tissue mosaicism, it is necessary to evaluate Y mosaicism in various tissues even in the case without Y chromosome on karyotyping.

Effect of Diethylnitrosamine and Diethanolnitrosamine on Serum Lactate Dehydrogenase in Rats

장성근,이동욱,박기현,Chang, Sung-Keun,Lee, Dong-Wook,Park, Ki-Hyon 생화학분자생물학회 1991 한국생화학회지 Vol.24 No.6

Diethylnitrosamine(DENA)과 Diethanolnitrosamine(DELNA)을 흰쥐에 투여하고 혈청에서 젖산 탈수소효소(LDH)의 활성도와 동위효소의 pattern을 관찰하였다. 이들의 투여로 인해 LDH의 총활성도는 크게 증가하되 약 2주일 후부터는 점차 감소되었다. 전기영동법에 의해 동위효소의 pattern을 측정한 결과 전형적인 5종의 isozyme만 확인되었고, 특히 LDH1의 상대적인 활성도는 정상쥐에 비해 크게 증가된 반면 LDH5는 오히려 다소 감소되었다. 그러나 이들 두 isozyme의 상대적인 활성도의 비는 크게 변화되지 않았다. 이와 같은 LDH isozyme 의 특이한 활성도의 변화는 nitrosamine화합물들에 의해 유발되는 생체손상의 한 특정인 것 같다. Lactate dehydrogenase(LDH) and its isozyme patterns were investigated in the serum of the rats treated with diethylnitrosamine(DENA) or diethanolnitrosamine(DELNA). LDH activity was remarkably increased by the treatment of these chemicals, but that was gradually decreased from about 2 weeks after administration of the chemicals. Especially the relative activity of LDH isozyme 1 was increased remarkably, whereas that of isozyme. 5 was slightly decreased from the 5th days. However, the activity ratios of the isozymes were not changed significantly with time course. This unique alterations in serum LDH activity might be provide a useful information for the detection of tissue toxicity induced by these chemicals.

신이식 후에 발생한 이상자궁출혈 환자에서 시행한 자궁경하 자궁내막 절제술의 효과에 관한 연구

정다정 ( Da Jung Chung ),박기현 ( Ki Hyun Park ),정경아 ( Kyung Ah Jeong ),신종승 ( Jong Seung Shin ),배상욱 ( Sang Wook Bai ),이병석 ( Byung Seok Lee ),조동제 ( Dong Jae Cho ),송찬호 ( Chan Ho Song ) 대한산부인과학회 2003 Obstetrics & Gynecology Science Vol.46 No.6

목적 : 신이식 후 발생한 이상자궁출혈의 치료로써 자궁경하 자궁내막 절제술의 효과를 알아보고자 하였다. 연구 방법 : 1999년 1월부터 2001년 12월까지 신촌세브란스병원 산부인과에 이상자궁출혈을 주소로 내원한 62명의 신이식 환자를 대상으로 자궁경하 자궁내막 절제술을 시행하였다. 시술 전 에스트라디올, 황체형성호르몬, 난포자극호르몬, 갑상선자극호르몬, 프로락틴을 측정하였다. 시술 후 6개월 간 추적 관찰하였으며 치료에 실패한 8명에서는 levono Objective : To assess the effectiveness and safety of hysteroscopic endometrial ablation as a surgical management of abnormal uterine bleeding developed in renal transplant patients. Methods : Data were collected retrospectively from 62 patients referred

터너 증후군 환자에서 총 경동맥의 혈관 내막 두께 측정을 통한 잠재성 동맥경화의 관계분석

정경아(Kyung Ah Jeong),박기현(Ki Hyun Park),김정연(Jeong Yeon Kim),송계영(Kye Young Song),안철우(Chul Woo Ahn),배상욱(Sang Wook Bai),이병석(Byung Seok Lee),조동제(Dong Jae Cho),송찬호(Chan Ho Song) 대한산부인과학회 2002 Obstetrics & Gynecology Science Vol.45 No.6

목적 : 본 연구는 터너 증후군 환자를 대상으로 고해상도 초음파를 통한 총 경동맥의 혈관 내막 두께를 측정하여 이들의 잠재성 동맥경화로 인한 심혈관 질환의 발생 위험을 예측하고 그 위험인자에 관하여 알아보고자 하였다. 연구 방법 : 총 18명의 터너 증후군 환자와 18명의 건강한 젊은 여성 대조군을 대상으로 이들의 임상적 특징과 생화학적 특성 및 총 경동맥의 혈관 내막 두께를 측정하여 비교하였고 이에 영향을 미칠 수 있는 여러 임상적, 생화학적 위험인자들과의 관계를 분석하였다. 결과 : 두 군의 평균 연령과 체질량지수는 차이가 없었는데 터너 증후군 환자는 통계학적으로 유의하게 키가 작고 (147.8±7.9 vs 160.3±5.9, p<0.001) 허리-둔부둘레비가 증가되어 있었다 (0.86±0.04 vs 0.78±0.04, p<0.001). 공복시 혈당 (90.1±9.9 vs 79.4±4.4 ㎎/㎗, p<0.001)과 인슐린 (9.5±3.0 vs 4.7±1.0 μIU/㎖, p=0.009), 총 콜레스테롤 (187.1±21.3 vs 154.8±21.8 ㎎/㎗, p=0.014), 저밀도지단백치 (111.3±10.0 vs 82.8±16.4 ㎎/㎗, p=0.009)는 터너 증후군 환자에서 유의하게 높았고 총 경동맥의 혈관 내막 두께는 통계학적으로 의의있게 증가되어 있었다 (0.61±0.09 vs 0.49±0.02 ㎜, p=0.002). 총 경동맥의 혈관 내막 두께에 영향을 미치는 요인은 터너 증후군, 허리-둔부둘레비, 공복시 혈당 및 인슐린치였다 (Coefficients of correlation: 0.720, p<0.001; 0.671, P<0.001; 0.445, p=0.020; 0.904, p<0.001). 결론 : 터너 증후군 환자들은 임상증상이 없는 젊은 여성에서도 잠재성 동맥경화로 인한 심혈관 질환의 발생 위험성이 정상 대조군보다 높으며 이에 영향을 미치는 가장 중요한 위험인자는 인슐린 저항 증가이다. Objective : This study was undertaken to determine whether women with Turner syndrome have greater subclinical atherosclerosis and evaluate the relationship to risk factors for atherosclerosis. Methods : 18 Women with Turner syndrome and 18 women as control group were measured the intima media thickness (IMT) of common carotid artery by B-mode ultrasound. We compared the IMT between cases and controls, and analyzed risk factors which affect the IMT. Results : There are no differences between the groups in age and body mass index (BMI). The height was shorter (147.8±7.9 vs 160.3±5.9, p<0.001) and the waist-hip ratio (WHR) was significantly increased in Turner syndrome (0.86±0.04 vs 0.78±0.04, p<0.001). Fasting blood sugar (FBS) (90.1±9.9 vs 79.4±4.4 ㎎/㎗, p<0.001), fasting insulin (9.5±3.0 vs 4.7±1.0 μIU/㎖, p=0.009), total cholesterol (187.1±21.3 vs 154.8 ±21.8 ㎎/㎗, p=0.014), and LDL (111.3±10.0 vs 82.8±16.4 ㎎/㎗, p=0.009) were significantly higher in Turner syndrome. Compare to control, the IMT was significantly increased in Turner syndrome (0.61±0.09 vs 0.49±0.02 ㎜, p=0.002). In the analysis of correlation between the IMT and clinical & biochemical characteristics, Turner syndrome status, WHR, FBS and fasting insulin were significantly affecting factors (Coefficients of correlation: 0.720, p<0.001; 0.671, P<0.001; 0.445, p=0.020; 0.904, p<0.001). Conclusion : These results suggested that women with Turner syndrome might have an increased risk of subclinical atherosclerosis and insulin resistance was most important risk factor.

Comparison of Characteristics of Hepatic Microsomal Cytochrome P450-dependent Monooxygenases from Snake and Rat

Ja Young Moon(문자영),Dong Wook Lee(이동욱),Ki Hyun Park(박기현) 한국생명과학회 1998 생명과학회지 Vol.8 No.6

폐경 후 호르몬 대체요법 2년 후 총 경동맥의 혈관 내막 두께 변화로 동맥경화에 미치는 효과를 본 연구

정경아 ( Kyung Ah Jeong ),박기현 ( Ki Hyun Park ),안철우 ( Chul Woo Ahn ),배상욱 ( Sang Wook Bai ),이병석 ( Byung Seok Lee ),조동제 ( Dong Jae Cho ),송찬호 ( Chan Ho Song ) 대한폐경학회 2003 대한폐경학회지 Vol.9 No.1

목적 : 본 연구는 폐경 후 호르몬 대체요법 후에 총 경동맥의 혈관 내막 두께를 추억 관찰하여 동맥경화에 미치는 효과를 알아보고자 하였다. 연구방법 : 102명의 폐경된 여성에서 호르몬 대체요법 전후에 초음파로 총 경동맥의 혈관 내막 두께를 측정하였다. 호르몬 치료는 평균 27.9±3.7개월동안 시행한 후에 추적 관찰하였다. 102명 중 36명은 sequential regimen(0.625㎎ conjugated equine estrogens : CEE/5㎎ MPA), 34명은 continuous regimen (0.625㎎ CEE/5㎎ MPA), 12명은 2.5㎎ tibolone, 10명은 oral estrogen regimen (0.625㎎ CEE), 10명은 transdermal estrogen regimen (50㎍ estradiol)을 투여받았다. 치료전후에 혈압 및 생화학적 검사로 총 콜레스테롤, 중성지방, 고밀도지단백, 저밀도지단백, lipoprotein (a), 공복시 혈당, 인슐린, fibrinogen과 plasminogen activator inhibitor-1을 각각 측정하여 비교하였다. 결과 : 폐경 후 호르몬 대체요법을 시행받은 후에 총 경동맥의 혈관 내막 두께는 통계학적으로 유의하게 감소하였다 (평균치 좌 0.69±0.18 vs 0.53±0.20 ㎜, p<0.001: 우 0.65±0.11 vs 0.48±0.17 ㎜, p<0.001, 최대치 좌 0.73±0.20 vs 0.63±0.23 ㎜, p=0.001; 우 0.68±0.12 vs 0.57±0.20 ㎜, p<0.001). 혈압은 변화가 없었고 총 콜레스테롤 (210.7±34.0 vs 202.8±30.9 ㎎/이,p=0.029), 저밀도지단백 (113.3±25.5 vs 80.5±24.7 ㎎/이, p=0.029), lipoprotein (a) (51.3±13.8 vs 39.7±13.2 ㎎/㎗, p=0.038)와 인슐린치 (6.8±2.6 vs 4.4±2.7 ??U/㎖, p=0.026)가 의미있게 감소되었으며 그 외 생화학적 검사 수치는 유의한 변화를 보이지 않았다. 총 경동맥의 혈관 내막 두께와 생화학적 검사 모두에서 호르몬 치료 방법에 따른 차이는 없었다. 결론 : 단기간의 폐경 후 호르몬 대체요법은 동맥경화의 진행에 있어서 예방적 효과가 있다. Objective : To assess the 2-year effects of postmenopausal hormone replacement therapy (HRT) on progression of atherosclerosis, measured as intima media thickness (IMT) in carotid arteries. Methods : This study included 102 postmenopausal women recruited from the general population. Blood pressure, biochemical charicteristics, and common carotid IMT with B mode ultrasonography were measured at baseline and 27.9±3.7 years-HRT : 36 patients; sequential regimen (0.625㎎ conjugated equine estrogens: CEE/5㎎ medroxyprogesterone acetate: MPA), 34 patients; continuous regimen (0.625㎎ CEE/2.5㎎ MPA), 12 patients; 2.5㎎ tibolone, 10 patients; oral estrogen regimen (0.625㎎ CEE), 10 patients; transdermal estrogen regimen (50㎍ estradiol). Results : After HRT, total cholesterol (210.7±34.0 vs 202.8±30.9 ㎎/㎗, p=0.029), LDL (l13.3±25.5 vs 80.5±24.7 ㎎/㎗, p=0.034), lipoprotein (a) (51.3±13.8 vs 39.7±13.2 ㎎/㎗, p=0.038), and fasting insulin (6.8±2.6 vs 4.4±2.7 ??U/㎖, p=0.026) were significantly decreased. Significant changes of the other biochemical characteristics and blood pressure could not be demonstrated. Common carotid IMT (average : Lt. 0.69±0.18 vs 0.53±0.20 ㎜, p<0.001, Rt. 0.65±0.11 vs 0.48±0.17 ㎜, p<0.001; maximum : Lt. 0.73±0.20 vs 0.68±0.23 ㎜, p=0.001, Rt. 0.68±0.12 vs 0.57±0.20 ㎜, p<0.001) was significantly decreased. There were no difference between the groups according to method of HRT. Conclusion : The findings of this study show that 2-year of HRT was effective in slowing progression of subclinical atherosclerosis in postmenopausal women.