• 생명과학회지
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• 제8권4호
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• pp.400-409
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• 1998

Formation of adduct was studied in benzo(a)pyrene(BP)- and doxorubicin(Dx)-treated human NC-37 cells and isolated nuclei. Major adducts formed were determined by fluorescence absorption spectrophotometery and DNA-lin-ked protein assay. When isolated nuclei were exposed to carcinogens BP and DMBA, and anticancer drugs m-AMSA, ellipticine and Dx, varying degrees of adduct formation occured between DNA-protein complex and these drugs. When the mixture was centrifuged 1.7 M sucrose solution, binding BP and DMBA appeared to be similar between the sediment and the supernatant. When the sediment was centrifuged again with 0.35% polymin-P, the amount of BP bound was 2-fold greater in the protein(1077$\pm$55cpm) than in DNA fraction (470$\pm$20cpm), whereas that of DMBA was 1.6-fold greater in the DNA than in protein fraction. In the case of m-AMSA, ellipticine and Dx, the amount of binding was slightly greater in supernatant than in sediment in centrifugation with 1.7 M sucrose, and more than 3 times greater in the DNA- than in protein- fraction in centrifugation with 0.35% polymin P. DNA fractions which associated with a subset of nonhistone chromosomal protein were isolated from NC-37 cells exposed to $^{3}$H-BP and $^{14}$C-Dx. They were separated into two distince components DNA-S and DNA-P by centrifugation with 2M Nacl chromatin extraction. The results indicated that the amount of $^{3}$H-BP bound was 6.0-fold greater in DNA-P as compared with DNA-S, while that of $^{14}$C-Dx binding appreaed to be 6.2-fold greater in DNA-S than in DNA-P fraction. When $^{3}$H-BP binding wasdetermined in the presence of cold Dx, the amount of binding was reduced only in the DNA-P fraction, indicating that the interaction between DNA and protein is decreased. Gene expression by these drugs, BP treated cells were increased to compare with nomal cells but reduced by treatment with BP-Dx. These results suggest that the protein moiety which tightly bound to DNA-P fraction may play an important role in the regulation of gene expression.

• 한국식품조리과학회지
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• 제11권4호
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• pp.351-355
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• 1995

용매(MeOH, EtOH, CHCI$_{3}$/MeOH Water)를 달리한 마추출물의 broiling과 panfrying의 조리법 변이원과 2-AF, benzo(a)pyrene, sodium azide 등의 화학적으로 유도된 순수변이원에 대한 억제효과를 검색하였다. 대사활성물질인 S9 mixture 무첨가시 조리법 변이원에 대한 변이원과 용매종류 및 변이원과 용매사이의 상호관계성은 유의적인 차이를 보였으나, S9 mixture 첨가시에는 변이원, 마, 용매종류들사이 에 유의성을 보였다. 특히 S9 mixture 첨가시 높은 억제효과를 보였던 산마 EtOH 추출물은 panfrying 변이원에 대한 억제효과는 컸으나 조리법 변이원에 포함되었을 것으로 예상되었던 benzo(a)pyrene에 대해서는 억제효과가 낮았다. 순수변이원로서 sodium azide에 대한 마추출물은 산마가 25.4%로, 9.1%인 재배마보다 높았으나 용매에 따른 유의성은 없었다. 2-AF는 마종류에 대한 유의성은 없었으나 Water가 26.2%, MeOH가 41.9%, EtOH가 45.0%, CHCI$_{3}$/MeOH가 84.0%로서 높은 억제효과를 보였다. 특히 산마의 CHCI$_{3}$/MeOH 추출물은 plate당 10 mg 농도에서 가장 높은 91.5%의 억제효과를 보였을뿐 아니라 plate당 1 mg 농도에서도 67%의 높은 억제효과를 나타내었다. 이상과 같이, 본 실험에 사용된 변이원 중 2-AF에 대한 산마 CHCI$_{3}$/MeOH 추출물이 단위 추출물당 억제활성이 가장 높은 것으로 나타났다.

• 한국환경과학회지
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• 제13권4호
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• pp.413-420
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• 2004

The objectives of present study were to investigate the effects of benzo[a]pyrene(BaP) on cytotoxicity, lipid peroxidation and antioxidant enzymes in rat hepatocyte primary culture. Primary cultures of rat hepatocytes were incubated for 24 hr, 48 hr or 72 hr in the presence of various concentrations (0, 10, 20, 30, 50 or 100 $\mu.$ M) of BaP. Cytotoxicity and cell viability were determined by measuring glutamic oxaloacetic transaminase(GOT) activity, lactate dehydrogenase(LDH) activity and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide(MIT) value. Lipid peroxidation was evaluated using thiobarbituric acid reactive substances(TBARS) assay. Effects on antioxidant system were determined by measuring glutathione peroxidase(GPx) activity, glutathione reductase(GR) activity and glutathione concentration. Activities of GOT and LDH, MTT value as well as TBARS concentration were not affected by up to 100 $\muM$ of BaP for 24 hr incubation. However, BaP at the concentration of 50 $\muM$ for 48 hr incubation or at the concentration of 30 $\muM$ for 72 hr incubation began to increase LDH activity and TBARS concentration but decrease MTT value, representing that BaP caused cytotoxicity and decreased cell viability in dose- and time-dependent manners. GPx activity began to be decreased by BaP at the concentration of 50 $\muM$ for 72 hr incubation. Whereas, GR activity began to be decreased by BaP at the concentration of 20 $\muM$ for 72 hr incubation. Glutathione concentration began to be decreased by BaP at the concentration of 20 $\muM$ for 72 hr incubation and was further reduced to 90% by 100 $\muM$ of BaP. These results demonstrate that BaP caused cytoctoxicity and decreased cell viability by increasing lipid peroxidation and decreasing glutathione concentration as well as activities of GPx and GR.

• 분석과학
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• 제36권6호
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• pp.291-299
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• 2023

식용유지 중 benzo[a]pyrene 추출법으로 액체-액체 추출법, Soxhlet 추출법, 초음파 추출법 등이 사용되고 있으나 이런 방법들은 추출 시간이 길고 많은 양의 용매를 사용하는 단점이 있다. 따라서 본 연구에서는 간편하고 단시간에 분석이 가능한 QuEChERS법을 적용하여 현재의 복잡한 benzo[a]pyrene 분석법을 개선하고자 하였다. 본 연구에서 적용한 QuEChERS법은 식용유지를 n-hexane saturated acetonitrile과 n-hexane으로 녹인 후 magnesium sulfate, sodium chloride를 이용해 추출하는 방식이다. 이 시험법에 대한 유효성 검증 결과, 검출한계와 정량한계는 각각 0.02, 0.05 ㎍/kg 이며, 직선성은 0.1~10 ㎍/kg 농도 범위에서 검량선 결정계수 (R²)는 0.99 이상이었다. 회수율과 회수율에 대한 상대표준편차는 각각 74.5~79.3 %, 0.52~1.58 % 였으며, 정확성 및 정밀성은 각각 72.6~79.4 %, 0.14~7.20%로 나타났다. 위의 결과들은 식품의약품안전평가원의 「식품 등 시험법 마련 표준절차에 관한 가이드라인 (2016)」 및 「시험법 마련 표준절차 AOAC 가이드라인 (2023)」을 모두 만족하였다. 따라서 본 연구에서 제시된 분석법은 기존 분석법 대비 상대적으로 간편한 전처리 방법으로 분석 시간과 용매 사용량을 각각 92 %, 96 % 정도 절감할 수 있었다.

• 한국동물학회지
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• 제31권2호
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• pp.142-146
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• 1988

PAH화합물의 하나인 benzo(a)pyrene의 쥐 태반 microsomes에 의한 대사활성화를 조사하였다. B(a)P대사물은 C18- $\mu$ Bondapak칼럼을 사용하여 고압액체크로마토그래피로 분석하였다. 그 결과 B(a)P를 투여한 쥐의 태반 microsomes에 의한 주 대사산물은 발암성이 강한 7,8-diol B(a)P였으며, 또한 적은량의 4,5-diol B(a)P와 quinone류가 검출되었다. 3-methyl-cholanthrene을 투여한 경우 hydroxy B(a)P 와 quinone화합물이 주 대사산물이었으며 Phenobarbital을 전처리했을 경우 7,8-diol B(a)P이 주 대사산물인 것으로 나타났다.

• Journal of Microbiology and Biotechnology
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• 제13권4호
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• pp.624-627
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• 2003

Benzo[a]pyrene (B[a]P) is an environmental pollutant that has been implicated in carcinogenesis. Saccharomyces cerevisiae was treated with B[a]P, and the responses of its cytochrome P450 (CYP) enzyme and DNA-damage checkpoint genes were examined through gene expression profiles using a reverse transcription polymerase chain reaction (RT-PCR). The DNA-damage checkpoint genes tested were the chk1 and pds1 genes, involved in a metaphase arrest, the swi6 gene targeted by G1 arrest, the pol2 gene related to S phase arrest, and the cln2 gene encoding a cyclin protein, all of which are based on rad9 and rad24. Among these genes, no noticeable effect was found when the cells were exposed to various concentrations of B[a]P. However, the transcriptional activity of CYP51 was significantly different when the cells were exposed to B[a]P. Accordingly, the present results indicate that cytochrome P450 plays a more significant role than DNA-damage checkpoint genes in the response of S. cerevisiae to B[a]P.

• Fisheries and Aquatic Sciences
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• 제9권2호
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• pp.64-69
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• 2006

Cytochrome P450 (CYP1A) gene expression in the liver and sex steroid levels in plasma were investigated in olive flounder (Paralichthys olivaceus) exposed to tributyltin (TBT) and benzo[a]pyrene (BaP). We constructed a cDNA library and cloned a 230-base sequence encoding partial CYP1A DNA. The CYP1A gene expression level was estimated using northern blotting. Hepatic CYP1A mRNA levels in fish injected with BaP at 10 mg/kg body weight (b.w.) increased for 48 h after injection. However, fish injected with both BaP and TBT at 10 mg/kg b.w. showed no significant changes in CYP1A mRNA level after 48 h. Plasma concentrations of testosterone and $17{\beta}$-estradiol were not significantly different in males and females injected with BaP and TBT. We suggest that TBT-induced suppression of BaP bioactivity should be interpreted with caution in biomonitoring field studies.

• Fisheries and Aquatic Sciences
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• 제5권1호
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• pp.28-31
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• 2002

Effects of cimetidine, a cytochrome P450 inhibitor, on the benzo[a]pyrene (BaP)-mediated cytochrome P450 induction and lipid peroxidation of liver in olive flounder, Paralichthys olivaceus, were investigated. Fish were fed either a cimetidine-supplemented diet or a cimetidine-free control diet once daily to satiation for 3 days. After 6 hrs of last feeding, the fish received intraperitoneal (i.p.) injection of BaP (20 mg/kg of body weight) dissolved in sterile corn oil $(100 \mu L)$ or received only a corn oil i.p. injection. At 1, 2, 3, and 7 days after the injection, hepatic cytochrome P450 and thiobarbituric acid reactive substances (TBARS), an indicator of lipid peroxidation, were analyzed. BaP injection resulted in an increase of hepatic cytochrome P450, and the fish fed the cimetidine-supplemented diet before injection of BaP showed delayed increase of hepatic cytochrome P450 compared to the fish fed a cimetidine-free diet and BaP injected. Injection of BaP clearly induced hepatic lipid peroxidation, and consistently higher TBAR values were shown in the fish fed a cimetidine­supplemented diet before injection of BaP than the fish injected with BaP alone.

• 한국지하수토양환경학회:학술대회논문집
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• 한국지하수토양환경학회 2006년도 총회 및 춘계학술발표회
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• pp.216-219
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• 2006

Water retention curve (WRC), one of soil hydraulic properties, is often approximated by property-transfer models (PTMs). Using the PTMs, we can estimate the WRCs from other physical properties such as particle-size distribution (PSD). The objective of this work was to investigate the performance of two PTMs with different origins for numerical simulations on transport of Benzo[a]pyrene in a soil. To do this, we chose both PTMs with different origins, i.e., (1) the lognormal distribution model (L anti NL models), and (2) the modified $Kov\'{a}cs$ model (MK model). The MK model showed tile worse performance in estimation of the WRCs. When transport of B[a]P was simulated, the MK model predicted to move farther than the L and NL models did, indicating that transport of B[a]P in a soil can be greatly influenced by the choice of PTMs.

• 한국식품위생안전성학회지
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• 제29권2호
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• pp.141-145
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• 2014

식용유지류의 섭취로 인해 발생하는 PAHs의 대표적인 발암물질인 벤조피렌의 안전성을 판단하기 위해 시중에 유통되는 288건의 식용유지제품에 함유된 벤조피렌 함량을 분석한 후 위해성을 평가하였다. 사용된 제품은 옥수수기름, 포도씨유, 올리브유, 들기름, 미강유, 참기름, 콩기름, 해바라기유 등이며 벤조피렌의 검출 범위는 불검출 ${\sim}4.78{\mu}g/kg$이었고 평균 농도는 $0.11{\mu}g/kg$로 나타났다. 국민건강영양조사 5기 2차년도(2011) 자료를 이용하여 전체인구집단과 섭취군의 식용유지류 섭취량을 산출하여 만성 1일 인체노출량을 평가한 결과 식용유지류 섭취로 인한 벤조피렌 노출량은 전체인구집단의 경우 $4.26{\times}10^{-3}ng/kg$ b.w./day, 식용유지류 섭취군의 경우 $7.64{\times}10^{-3}ng/kg$ b.w./day로 나타났다. 인체 위해정도를 파악하기 위해 벤조피렌의 mouse 위상부암에 대한 $BMDL_{10}$값인 0.31~0.74 mg/kg b.w./day와 만성 1일 인체노출량을 이용하여 산출한 MOE 값은 전체인구집단에서는 $7.28{\times}10^7{\sim}1.74{\times}10^8$, 섭취자군에서는 $3.95{\times}10^7{\sim}9.42{\times}10^7$ 수준이었다. 따라서 식용유지류 섭취로 인한 벤조피렌 인체노출의 위해정도는 매우 낮은 것으로 판단된다.