단백질 정량 발색반응에 미치는 Zinc Protoporphyrin의 영향

김미리,최보희,김현정,조은비,최현아,홍정일 서울여자대학교 자연과학연구소 2012 자연과학연구논문집 Vol.24 No.-

The Bradford and bicinchoninic acid (BCA) assays are commonly-used colorimetric methods for quantifying amount of proteins. In the present study, effects of zinc protoporphyrin (ZnPP) on color responses of these assay methods were investigated. ZnPP and zinc did not affect the color response of Coomassie brilliant blue dye in the Bradford assay. ZnPP, however, reacted with the BCA reagent and increased the color intensity in the BCA method concentration-dependently. In the current assay system, the color response by 20 µM of ZnPP was comparable to that of ~50 µg/mL of standard protein of bovine serum albumin. Zinc did not affect the BCA assay either. Color intensity of the BCA method was also enhanced in the cell lysates containing ZnPP. The present results indicate that various factors could affect the color response of protein assays, and ZnPP is one of the interfering factors in the BCA method, which is considered carefully in related studies.

Curcumin과 Protoporphyrin류 화합물의 상호작용에 의한 형광특성 변화

김현정,김미리,최보희,홍정일 서울여자대학교 자연과학연구소 2013 자연과학연구논문집 Vol.25 No.-

Curcumin, a dietary polyphenol from Curcuma longa L., has been reported to show many beneficial bioactivities. Protoporphyrins are widely distributed in various natural sources. They are also intermediates in the endogenous synthesis of heme and commonly form a complex with metal ions. In the present study, we investigated changes in fluorescence properties of curcumin and protoporphyrin IX (PPIX) through their interactions. Fluorescence intensity of curcumin increased concentration-dependently showing a peak emission at 535 nm with an excitation at 440 nm. Curcumin fluorescence was not changed by PPIX; the fluorescence intensity of PPIX significantly decreased in the presence of curcumin. Curcumin fluorescence decreased time-dependently; the fluorescence from PPIX was rather enhanced according to time, which was less pronounced in the presence of curcumin. Curcumin fluorescence in cells decreased markedly, which is slightly enhanced by co-treatment of PPIX. Curcumin fluorescence also decreased by zinc protoporphyrin; the fluorescence from ZnPP was also quenched by curcumin. The present results suggest a possible interaction of metal porphyrins with curcumin, which may influence on their bioactivities.

빛, 용매와 zinc protoporphyrin에 의한 MTT 포마잔의 화학적 동태 변화

김주현,홍정일 한국식품과학회 2018 한국식품과학회지 Vol.50 No.1

The MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay is commonly used for analyzing the cell viability. In this study, effects of various solvents, different lights, and zinc protoporphyrin (ZnPP) on the chemical behavior of MTT formazan were investigated. The color response of MTT formazan in NaOH was highly pronounced; the absorbance of MTT formazan in 0.1 N NaOH at 550 nm was >2-fold higher than that in water, dimethyl sulfoxide (DMSO), methanol, and ethanol. MTT formazan in DMSO and NaOH (>0.1 N) was relatively stable under fluorescent and UV light at 365 nm; its rapid degradation was induced under UV light at 254 nm in all solvents. ZnPP degraded MTT formazan under light in a time- and concentration-dependent manner; MTT formazan in 0.1 N NaOH was the most sensitive to ZnPP, followed by DMSO. These results suggest that NaOH and DMSO might be suitable media for MTT formazan for monitoring photosensitizing properties.

우리나라 도시와 농촌 지역 주민의 혈중 연 및 혈중 Zinc Protoporphyrin농도 비교 연구

김준연,김병권,홍영습,이용언,정갑열 동아대학교 부설 산업의학연구소 1997 산업의학연구소 논총 Vol.- No.3

The study was carried out on healthy population in urban and rural area, in order to compare the level of blood lead (PbB) and blood zinc protoporphyrin (ZPP) between the population living in each area from July 1 1994 to December 31 1994. We stratificated the results with sex, age and area. The results obtained were as follows; 1. In urban area, overall mean of the PbB level was 7.08±1.61㎍/dl and both men and women had the highest level of PbB, which were 8.15±0.85㎍/dl, 6.90±1.48㎍/dl, in less than 39 years old group. In rural area, overall mean of the PbB level was 6.75±1.61㎍/dl and both men and women had the highest levels of PbB, which were 7.25±2.29㎍/dl, 7.00±2.92㎍/dl in 40∼49 years old group. And there was statistically significant difference between mean of the men's PbB level in urban area and that in rural area (p<0.05). 2. In urban area, overall mean of the ZPP level was 32.08±8.28㎍/dl and men had the highest level of ZPP, which was 29.59±5.13㎍/dl, in 40∼49 years old group, and women had that, which was 36.83±7.95㎍/dl, in less than 39 years old group. In rural area, overall mean of the ZPP level was 34.54±11.31㎍/dl and men had the highest level of ZPP, which was 32.10±5.45㎍/dl, in less than 39 years old group and women had that, which was 40.17±14.02㎍/dl, in 40∼49 years old group. And there was statistically significant difference between mean of the men's ZPP level in urban area and that in rural area (p<0.05). 3. There was statistically significant difference in PbB and ZPP between men and women group in both area (p<0.05) but not significant among age group in both area (p>0.05).

연 폭로시 성별에 따른 혈중 Zinc Protoporphyrin의 양-반응 관계

김용배,리갑수,황규윤,이성수,안규동,이병국,안현철,Kim, Yong-Bae,Lee, Gap-Soo,Hwang, Kyu-Yoon,Lee, Sung-Soo,Ahn, Kyu-Dong,Lee, Byung-Kook,Ahn, Hyun-Cheol 대한예방의학회 1999 Journal of Preventive Medicine and Public Health Vol.32 No.4

Objectives : To evaluate whether a relation between blood lead and zinc proto porphyrin(ZPP) was modified by gender in Korean lead workers. Methods : A cross-sectional study was conducted with 1,304 male and 101 female subjects in 1997. The relation between blood lead and ZPP were evaluated with linear, exponential, and quadratic models. Then, the different effect of gender on the relation was examined by adding the interaction terms in the each model. Results : $Mean{\pm}SD$ of blood lead and ZPP level was different between male$(27.7{\pm}10{\mu}g/dl\;and\;51.3{\pm}23.4{\mu}g/dl)$ and female subjects$(22.5{\pm}9.2{\mu}g/dl\;and\;78.7{\pm}38.6{\mu}g/dl)$. After adjusting for possible confounders, the effect modification by gender was significant in linear$(\beta=1.119,\;p<0.001)$, exponential$(\beta=Exp(0.008),\;p<0.05)$, and quadratic model$(\beta=1.388,\;p<0.001)$. In separate analysis, a quadratic relation between blood lead and ZPP was shown in male lead workers$(\beta=0.036,\;p<0.001)$, but an exponential relation in female lead workers$(\beta=Exp(0.029), p<0.001)$. Conclusion : Our data showed that the increasing rate of ZPP in female were always higher than in male lead workers, suggesting that females were more susceptible to occupational lead exposure than males.

MTT formazan의 발색에 미치는 zinc protoporphyrin의 영향

박경아(Kyung A Park),최현아(Hyun A Choi),김미리(Mi-Ri Kim),최유미(Yoo Mi Choi),김현정(Hyun Jung Kim),홍정일(Jungil Hong) 한국식품과학회 2011 한국식품과학회지 Vol.43 No.6

본 연구는 세포의 사멸 및 성장변화 등의 평가에 널리 이용되는 MTT assay에서 생성된 formazan dye에 미치는 ZnPP의 영향을 조사하였다. ZnPP는 생체 내에 자연적으로 생성되거나 다양한 관련 실험에 인위적으로 첨가해주는 물질로서, MTT formazan dye의 빛에 의한 탈색을 가속화시키는 것으로 밝혀졌다. Formazan dye의 분해는 5와 10 μM ZnPP 존재 시 반감기를 기준하여 각각 10 및 20배 가량 가속화되었으며, 빛이 차단된 조건에서는 영향을 미치지 않았다. ZnPP 구조 중 Zn는 formazan dye의 탈색에 영향을 미치지 않았으나, porphyrin 구조와 공통적인 감광체 성질을 나타내는 MB에 의해서 ZnPP 존재 시와 유사하게 dye의 탈색을 가속화 시켰다. 이러한 ZnPP와 MB에 의한 formazan dye의 탈색반응은 NAC와 β-carotene에 의해 지연되었으나, BHT에 의한 저해효과는 나타나지 않았다. 본 결과는 세포 중에 존재하는 ZnPP 등의 감광체류가 MTT tetrazolium으로부터 환원된 formazan dye의 빛에 의한 신속한 분해를 유도하며, 관련 실험에서 빛의 차단 등의 조치와 함께 각별히 유의해야함을 시사한다. Zinc protoporphyrin (ZnPP) is produced endogenously during heme metabolism and treated in cells as a heme oxygenase inhibitor. In the present study, the effects of ZnPP on the color response of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, a commonly-used method for analyzing cell viability, were investigated. ZnPP induced rapid decolorizaion of MTT formazan under light; the degradation rates were 10- and 20- folds faster in the presence of 5 and 10 μM ZnPP, respectively. Methylene blue (MB), another type of photosensitizer, also accelerated degradation of formazan under light. Butylated hyroxytoluene did not inhibit ZnPP- or MB-induced formazan degradation. The color degradation of formazan dye was signficantly delayed in the presence of N-acetylcysteine or β-carotene. The present results suggest that certain photosensitizing compounds may affect the color and stability of MTT formazan, which should be carefully considered when conducting the MTT assay.

Protective effects of Korean herbal remedy against oxidative stress in cardiomyocytes

Kim, Myung-Sunny,Kwon, Dae Young,Cho, Hye-Jin,Lee, Myeong Soo John Wiley Sons, Ltd. 2006 Phytotherapy research Vol.20 No.3

<P>Ondamtanggagambang (ODG) has been used as a prescription for psychological anxiety and depression in Korean medicine. In this study, we found that ODG have protective effects against oxidative stress in cardiomyocytes. Pretreatment with ODG extract prevented H<SUB>2</SUB>O<SUB>2</SUB> and ZnCl<SUB>2</SUB>-induced cell damage in H9c2 cardiomyocytes, whereas simultaneous treatment of ODG extract did not. The protective effect of ODG extract on oxidative stress-induced damage was suppressed significantly by heme oxygenase (HO) inhibitors, zinc protoporphyrin-IX (ZnPP-IX, p < 0.01) and tin protoporphyrin-IX (SnPP-IX, p < 0.01) in H9c2 cells. ODG stimulation of cells strongly induced the expression of HO-1 protein. Taken together, it is suggested that ODG-induced expression of HO-1 may have a beneficial role in cardiomyocytes under oxidative stress. Copyright © 2006 John Wiley & Sons, Ltd.</P>

Evaluation of Erythrocyte Zinc Protoporphyrin/Heme Ratio for Iron-Deficiency Anemia in Pregnant Korean Women

Tchai, Bum Suk,Kwon, Seok Woon,Han, Jung Ho Seoul National University 1993 The seoul journal of medicine Vol.34 No.3

직업적 연폭로에서 혈중 연량 수준에 따른 선별검사로서의 혈중 ZPP 검사의 타당도

한구석,안현철,김용배,리갑수,김화성,황규윤,장봉기,이성수,안규동,이병국 순천향대학교 산업의학연구소 1999 순천향산업의학 Vol.5 No.1

In order to evaluate the validity of current screening criteria of lead exposure and to provide effective information for the better health management of lead workers, author investigated the interrelationship between blood lead and blood zinc protoporphyrin(ZPP) and evaluate the validity of blood zinc protoporphyrin as the screening test and blood lead as gold standard of diagnostic test. The total of 2245 lead workers (male workers: 1982, female workers : 263) from 12 lead using industries were studied. Blood lead, ZPP, hemoglobin and hematocrit were selected as lead exposure variables. Personal information on sex, age and work duration were also checked. Blood lead was measured with flameless atomic absorption spectrometry (model: Hitachi 8100) using standard addition method. Blood ZPP was measured with portable hematoflurometer (model: Aviv 206). The results obtained were as follows: 1. The mean blood lead of all lead workers were 0.7±14.3㎍/㎗(Male: 31.0±14.4㎍/㎗, Female: 28.2±12.9㎍/㎗), whereas the mean blood ZPP of those were 68.1±47.5㎍/㎗(Male: 63.9±43.7㎍/㎗, Female: 100.1±61.0㎍/㎗). The mean value of hemoglobin and hematocrit of all lead workers were 14.5±1.5g/㎗(Male: 14.7±1.3g/㎗, Female: 12.3±1.2g/㎗) and 45.4±3.6(Male: 46.2±2.8, Female: 39.5±3.1) respectively. 2. Validity of test using blood zinc protoporphyrin as screening test for blood lead levels of 30㎍/㎗ as gold standard of diagnostic test revealed that the highest validity value for male and female lead workers were 50 and 90 ㎍/㎗ of ZPP respectively, showing 50 and 60 ㎍/㎗ of ZPP for all lead workers. While those test for blood lead levels of 40㎍/㎗ as gold standard of diagnostic test revealed that the highest validity value for male and female lead workers were 60 and 90 ㎍/㎗ of ZPP respectively, showing 60 ㎍/㎗ of ZPP for all lead workers, those test for blood lead levels of 50 and 60㎍/㎗ as gold standard of diagnostic test revealed that the highest validity value for male and female lead workers were 60 or 70 and 100 ㎍/㎗ ZPP respectively, showing 60and 70 or 80 ㎍/㎗ of ZPP for all lead workers. 3. Predictive value of test using blood zinc protoporphyrin as the screening test for different blood lead levels of 30, 40, 50 and 60㎍/㎗ as gold standard of diagnostic test revealed that positive predictive value of screening test were decreased by the increase of blood lead level regardless of csreening criteria value ZPP and negative predictive value of screening test were increased accordingly regardless of screening criteria value of ZPP. On the other hand, within same blood lead level as a gold standard positive predictive values were inceased by the increase of screening criteria value of ZPP, whereas the negative predictive value were decreased accordingly by the the increase of screening criteria value of ZPP. 4. The correlation analysis of all lead workers among study variables revealed that CPbB and natural log transformation ZPP(LZPP) were better correlated with other variables and found to be better estimate than PbB and ZPP themselves. The age was correlated with all other variables, but the work duration was not. The correlation analysis of male and female workers among study variables revealed the same results with all lead workers. 5. The slope of simple linear regression of CPbB (independent variable) with ZPP in female workers(3.223)was steeper than in male workers(2.144). That of lead workers with work duration of less than 1 year was steeper than that of lead workers whose work duration was more than 1 year. But there was no slope difference between two groups divided by their work duration of 2 years less and more. The slope of simple linear regression of CPbB lead (independent variable)with ZPP in lead workers whose blood lead were higher than 30㎍/㎗and 40㎍/㎗ were steeper than those whose blood lead were less than 30㎍/㎗ and 40㎍/㎗. 6. The slope of simple linear regression of CPbB (independent variable) with ZPP in lead industries with low mean blood lead were less steeper than those with high mean blood lead. 7. The slope of simple linear regression of CPbB (independent variable) with LZPP in female workers(0.0308was steeper than in male workers(0.0253). That of lead workers with work duration of less than 1 year was steeper than that of lead workers whose work duration was more than 1 year. But where was no slope difference between two groups divided by their work duration of 2 years less and more. While the slope of simple linear regression of CPbB lead (independent variable) with LZPP in lead workers whose blood lead were hogher than 30㎍/㎗ was not steeper than those whose blood lead were less than 30㎍/㎗, there was no slope difference between two groups whose blood lead were less than 40㎍/㎗ and more than 40㎍/㎗. 8. The slope of simple linear regression of CPbB (independent variable) with LZPP in lead industries with low mean blood lead were less steeper than those with high mean blood lead, but the difference was not significant then that of CPbB and ZPP. With above results, it is recommended that the screening criteria value of ZPP should be down to 60-70㎍/㎗ from 100㎍/㎗ for male lead workers and screening criteria of ZPP for female lead workers was all right with current value of 100㎍/㎗. So it is recommended to make different screening criteria of ZPP for male and female workers like the hemoglobin and hematocrit screening criteria. It is also preferable to use CPbB and LZPP rather than uncorrected PbB and ZPP in the evaluation of lead exposure for lead workers.

건강한 일부 도시지역 주민의 혈중 연 및 Zinc protoporphyrin 농도

정갑열,김준연,김동일,장형심,김용규,최안홍,이영호,김정만 동아대학교 부설 산업의학연구소 1995 산업의학연구소 논총 Vol.- No.2

1991년 5월 1일부터 약 6개월 동안 남자 1,193명 및 여자 658명을 대상으로 우리나라 도시 거주자들의 혈중 혈색소치, ZPP치 및 연농도를 성별, 연령별로 측정한 결과 아래와 같은 결과를 얻었다. 1. 혈색소의 평균치는 남자 14.55±1.82g/dl, 여자 12.61±1.18g/dl이었으며, 20세 이상 군에서는 남자가 여자보다 유의하게 높았다(p<0.05). 2. 혈중 ZPP치는 32.54±9.88(32.09~32.99㎍/dl, 95% C.I.)㎍/dl이었으며 7세 미만 소아의 혈중 ZPP치는 남아 및 여아가 37.49±13.31㎍/dl 및 35.77±11.85㎍/dl로 7세 이상군의 남자 및 여자의 31.91±8.23㎍/dl 및 30.11±9.11㎍/dl보다 유의하게 높았다(p<0.05). 3. 혈중 연농도는 23.84±4.58(23.55~24.14, 95% C.I.)㎍/dl이었으며, 7세 미만 소아의 혈중 연농도는 남아 및 여아가 25.10±5.21㎍/dl 및 24.45±4.18㎍/dl로 7세 이상군의 남자 및 여자의 24.28±3.00㎍/dl 및 21.99±5.05㎍/dl보다 유의하게 높았다(p<0.05). 이상의 결과를 요약하면 우리나라 도시 거주인은 남녀 모두 전연령에 걸쳐 높은 혈중 ZPP치 및 연농도를 나타내었으며 이러한 현상은 학동기전 소아에서 더욱 현저하므로 이들에 대한 관리대책 수립이 필요하다고 사료된다. Hemoglobin(Hb), zinc protoporphyrin(ZPP) and blood lead(PbB) levels were determined for 1,851 blood samples collected from healthy urban population to establish reliable baselines for Hb, ZPP and PbB levels by age and sex. ZPP values were analyzed with a hematofluorometer and PbB determinations were concurrently carried out using flameless atomic absorption spectrophotometry. The blood sampling period was about 6 months from May, 1991 and the summarized results were as follows: 1. The mean value of Hb in male and female were 14.55±1.81g/dl and 12.6±1.18g/dl respectively and there was statistically significant defference(p<0.05). 2. The mean value of ZPP in pre-schoolchildren was 37.49±13.31㎍/dl for male, 35.77±11.85㎍/dl for female and that of ZPP in after 7 years groups was 31.91±98.23㎍/dl for male, 30.11±9.11㎍/dl for female and there was statistically significant difference(p<0.05). 3. The mean value of PbB in pre-schoolchildren was 25.10±5.21㎍/dl for male, 24.45±4.18㎍/dl for female and that of PbB in after 7 years groups was 24.28±3.00㎍/dl for male, 21.99±5.05㎍/dl for female and there was statistically significant difference(p<0.05).