• Journal of Ginseng Research
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• 제16권3호
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• pp.210-216
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• 1992

The effect of saponin extracted from Panax grneng CA Meyer on DNA repair and replicative DNA synthesis were examined in CHO-Kl cells cotreated with benzo(a)pyrene and rat liver S-15 fraction. The DNA strand breaks inititated by benzo(a)pyrene metabolites were measured by alkaline election technique. The addition of ginseng saponin to the culture media resulted in decrease of benzo(a)pyrene-induced DNA strand breaks, and restored the suppressed-semiconservative-DNA-synthesis by the carcinogen. DNA repair synthesis in the damaged cells was also elevated by the ginseng treatment when the repairing activites were measured for the (3H)-thymidine incorporation into the carcinogen damaged cellular DNk Comparative analysis of DNA-adduces of benzo(a)pyrene metabortes in microsomes suggested that ginseng saponin treatment in rats reduced the formation of electrophilic metabolites of benzo (a)-pyrene in the rat liver microsomes.

• 한국환경보건학회지
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• 제40권1호
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• pp.55-62
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• 2014

Objectives: $^{32}P$-postlabeling assay is the most sensitive method of detecting DNA adduct formation. However, it is limited by a low sample throughput and use of radioisotopes (RI). In this study, we modified it to minimize these limitations and applied it to Z. platypus exposed to Benzo(a)pyrene (BaP) in order to investigate DNA adduct formation (effect biomarker for pollutants) in Z. platypus for assessing risk of waterborne BaP exposure. Methods: DNA hydrolysis was performed only with Micrococcal nuclease (MNase), RI reduction test was performed and the overlapping steps between thin layer chromatography (TLC) and radioisotope high-performance liquid chromatography (RI-HPLC) were omitted. The application of a modified method to Z. platypus exposed to BaP was performed. Results: The results revealed that the amount of RIs used can be reduced roughly 10-fold. Because the analysis time was shortened by 8.5 hours, the sample throughput per hour was increased compared with the previous method. The results of applying modified $^{32}P$-postlabeling assay to Z. platypus, DNA adduct formation in Z. platypus showed dose-dependency with the BaP concentration. Only BPDE-dGMP was detected as a DNA adduct. Conclusion: These results demonstrate that the modified $^{32}P$-postlabeling assay is a suitable method for detecting DNA adduct formation in Z. platypus exposed to waterborne BaP and will be useful in risk assessment of carcinogenic effect in aquatic environment due to BaP.

• Journal of Ginseng Research
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• 제13권1호
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• pp.42-48
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• 1989

PAH 계 화합물들의 Bay region diol epoxide 들의 target tissue에 대한 결합은 암유발과 관련 되어 있다. 본 논문에서는 ICR mice의 간에서의 BP-DNA-adduct 생성에 미치는 poly acetylene 화합물인 panaxynol 과 panaxydol 의 효과를 조사하였 다. Panaxynol 과 panaxydol 을 전처리한 ICR mice 의 간 마이크로좀을 포함하는 incubation system 은 calf thymus DNA 에 대한 BP binding을 뚜렷이 감소시켰다. [$^3H$]-BP($300\mu$Ci/21nmoles/0.1ml DMSO. i. v. ) 즐 mice 에 주사 후 24시간 후에 간 DNA 에서의 방사능을 측정하였다. HPLC 에 의해 cochromatography 한 두개의 standard marker (acetophenone. bytyrophenone)을 사이에 나타나는 DNA adduct 들을 잠정적으로 확인한 결과 (-) BP-7.8-diol로부터 생성되는 major adduct 인 (+) BP-diol epoxide I: dGuo adduct (peak II)는 대조군보다 약 22% 감소된 반면에 minor adduct 인 (-) BP-diol epoxide I: dGuo adduct (peak III)는 대조군의 69%로 감소되었다. 그리고 (+) BP-7, 8-diol로부 터 생성되는 minor adduct 인 BP-diol epoxide I II : Guo adduct (peak IV)는 대조군의 58%로 감소되었다. 이러한 결과는 panaxydol이 ($\pm$) B BP-7,8-diol로부터 일반적으로 생성되는 adduct들 중 major보다는 minor adduct들의 생성에 더 많이 관석했음을 보여준다.

• BMB Reports
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• 제28권1호
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• pp.1-5
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• 1995

The effect of 6-sulfooxymethyl benzo(a)pyrene (SMBP) on conformational changes of calf thymus DNA was investigated. As SMBP is a strong electrophile, the covalent binding of SMBP to DNA should distort three dimensional conformation of DNA at the binding sites. A formaldehyde-unwinding methods were used to determine the rate of DNA denaturation. The increase in absorbance at 251nm was detected by addition of formaldehyde following treatment with SMBP. SMBP changed supercoiled DNA to relaxed and linear DNA as determined by electrophoresis, which was similar to the change in DNA due to in vitro treatment with benzo(a) pyrene diol epoxide. Treatment with SMBP completely denatured DNA under alkaline conditions. However, DNA was nicked or partially denatured under neutral condition. The absorption band of DNA was increased by the treatment with SMBP in V79 cells, which may be explained by the formation of stabilized SMBP-DNA adduct.

• Journal of Ginseng Research
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• 제13권1호
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• pp.37-41
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• 1989

Benzo(a)pyrene(BP)의 monooxygenase(AHH)에 의해서 생성된 반응성 대사물질들의 in vitro DNA와의 결합 및 BP 대사에 관여하는 효소들의 활성도에 미치는 인삼 물추출물의 영향을 조사하였으며, DNA-BP metabolite adduct들은 Sephadex LH-20 column으로 chromatography하여 5개의 major peak 들을 얻었다. 이 peak 들을 극성이 큰 순서대로 A부터 E까지 임의로 정하고 5개의 peak들을 7,8-diol-9,10-oxide(A), 7,8-oxide(B). 4,5-oxide(C), 9-HO-BP(D & E) adduct들로 잠정적으로 확인하였다. Peak A, C, D 그리고 E는 각각 대조군의 30, 15, 20 그리고 30%로 감소되었으며 peak B는 의미있는 변화를 보이지 않았다. DNA-BP 결합 억제와 관련하여 in vitro와 in vivo 투여시의 경향이 유사하여 EH의 활성도만 BP투여 대조군보다 38%정도 의미있게 유도되었다.

• 생약학회지
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• 제34권4호통권135호
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• pp.297-302
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• 2003

Cnidii Rhizoma water extract (CRW) was tested for liver cancer chemopreventive potential by measuring the inhibition of phase I enzyme and benzo[a]pyrene-DNA adduct formation and induction of phase II detoxification enzymes. There was 17.0% inhibition in the activity of cytochrome P450 1A1 enzyme with the treatment of 150 mg/ml CRW. At concentration of 30 mg/ml CRW, the binding of $[^3H]B[a]P$ metablites to DNA of NCTC-clone 1469 cell was inhibited by 33.3%. CRW was potent inducer of quinone reductase (QR) and glutathione S-transferase (GST) activities in cultured murine hepatoma Hepalc1c7 cells. However, hepatic glutathione (GSH) level was not influenced by CRW. These findings suggest that CRW has chemopreventive potential of liver cancer by inhibiting cytochrome P450 1A1 activity and benzo[a]pyrene-DNA adduct formation and inducing QR and GST activities.

• Korean Journal of Acupuncture
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• 제19권1호
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• pp.7-13
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• 2002

1. 천궁약침액은 1${\times}$에서 15.1%의 cytochrome P4501A1 저해효과를 나타냈으며 그 억제효과는 약침액의 농도가 높을수록 증가하였다. 2 천궁약침액은 benzo[a]pyrene과 세포의 DNA 결합을 유의성있게 억제시켰다. 이상의 결과를 종합해 볼때 천궁약침액은 효과적으로 cytochrome P4501A1 효소를 저해했으며, 발암물질과 DNA의 결합을 억제시켜 외부 물질 또는 대사물에 의해 일어날 수 있는 돌연변이와 암 발생을 억제할 것으로 사료된다.

• 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.

• 한국독성학회:학술대회논문집
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• 한국독성학회 2002년도 국제심포지움
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• pp.20-45
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• 2002

In this study, we demonstrated the in vitro and in vivo formation of carcinogen-lipid adduct and its correlation with DNA or protein adducts. The lipids from serum or hepatocyte membranes of Spragu-Dawley rats. human serum, and standard major lipids were in vitro reacted with benzo[a]pyrene(BP) and BP metabolites. 7,8-Dihydroxy-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]-pyrene(BPDE-I), an ultimate carcinogenic form of BP, was covalently bound to triglyceride(TG). BPDE-I-TG adducts isolated by thin-layer chromatography (TLC) were further detected by high performance liquid chromatography(HPLC). TGs, including triolein, tripalmitin and tristearin, showed positive reactions with BPDE-I. However, cholesterol, phospholipids(Phosphatidylcholine, phosphatidyl-ethanolamine, phosphatidyl-inositol and sphingomyelin) and nonesterified fatty acids(palmitic acid, oleic acid, linoleic acid and stearic acid) did not react with BPDE-I. In addition, other BP metabolites (BP-phenols and -diols) did not react with TG, which TG appeared to be the most reactive lipid yet studied with respect to its ability to form an adduct with BPDE-I. There was a clear-cut dose-respect to its ability to form an adduct with BPDE-I-lipid adduct in vitro between TG and [1,3-3H]BPDE-I. In an animal study, BPDE-I-TG was also formed in the serum of rats orally treated with BP(25 mg/rat). Also, obvious correlations between [3H]BP related-biomolecule adducts (DNA, protein) or lipid damage and the BPDE-I-TG adduct were obtained in various tissues of mice i.p. treated with [3H]BP. These data suggest that TG can form an adduct with BPDE-I, as do other macromolecules (DNA, RNA, and protein). Therefore, a carcinogen-lipid adduct would be a useful biomarker for chemical carcinogenesis research and cancer risk assessment.

• 한국환경독성학회:학술대회논문집
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• 한국환경독성학회 2000년도 추계심포지움 및 학술발표회:바이오모니터링 기법을 이용한 환경위해성 평가
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• pp.74-74
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• 2000