Buplueri Radix의 1,2,4-benzentriol에 의해 유발된 DNA Damage에 대한 보호효과에 대한 연구
Protective Effect of Buplueri Radix(BR) Against 1,2,4-benzentriol Induced DNA Damage in Human Lymphocytes
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Objectives:Buplueri Radix(BR), used medical plant in Korea traditional medicine, contains various compounds, including a series of triterpene saponins known as saikosaponins. We performed this study for the protective effect of BR against oxidative damage induced by 1,2,4-benzentriol(BT) in human lymphocytes. Methods:In order to investigate the protective effect of BR against carcinogens, genotoxicity induced by benzene metabolite, BT were performed using cytokinesis-block micronucleus(CBMN) assay and comet assay. Results:The frequency of micronucleus at 25, 50 and 100μM concentration of BT were 8±2.36, 23±2.31, 35±4.17 respectively. In addition of BR with concentration of 25 and 50μg/mL, MN frequencies were significantly decreased. According to comet assay, BT induced DNA damage in a dose-dependent manner at concentration of 10 and 50 while BT with BR treatment decreased DNA breakage. No genotoxicity was observed by BR(25～50μg/mL) treatment alone on DNA breakage. Since BT can induce DNA damage through the generation of reactive oxygen species(ROS), we examined the level of ROS in human lymphocytes treated with BT and/or BR using DCF-DA, ROS-sensitive probe. The generation of ROS in BT-treated cells was also observed, and BR addition inhibited the level of BT-induced DNA damage. Conclusions:From above results it is suggested that BR could protect the cell and DNA from pro-oxidant effect by ROS by BT
Abstract
Objectives:Buplueri Radix(BR), used medical plant in Korea traditional medicine, contains various compounds, including a series of triterpene saponins known as saikosaponins. We performed this study for the protective effect of BR against oxidative damage induced by 1,2,4-benzentriol(BT) in human lymphocytes. Methods:In order to investigate the protective effect of BR against carcinogens, genotoxicity induced by benzene metabolite, BT were performed using cytokinesis-block micronucleus(CBMN) assay and comet assay. Results:The frequency of micronucleus at 25, 50 and 100μM concentration of BT were 8±2.36, 23±2.31, 35±4.17 respectively. In addition of BR with concentration of 25 and 50μg/mL, MN frequencies were significantly decreased. According to comet assay, BT induced DNA damage in a dose-dependent manner at concentration of 10 and 50 while BT with BR treatment decreased DNA breakage. No genotoxicity was observed by BR(25～50μg/mL) treatment alone on DNA breakage. Since BT can induce DNA damage through the generation of reactive oxygen species(ROS), we examined the level of ROS in human lymphocytes treated with BT and/or BR using DCF-DA, ROS-sensitive probe. The generation of ROS in BT-treated cells was also observed, and BR addition inhibited the level of BT-induced DNA damage. Conclusions:From above results it is suggested that BR could protect the cell and DNA from pro-oxidant effect by ROS by BT
참고문헌 (30)
- Martyn T.S., "The mechanism of benzeneinduced leukemia : A hypotheis and speculations on the causes of leukemia. Environ. Healt", Perspect, 6, pp.1219~1255 (1996), A hypotheis and speculations on the causes of leukemia.Environ.Healt.Perspect.104(6),1219-1255
- Li A.S., "DNA breakage induced by 1,2,4- benzenetriol : relative contributions of oxygenderived active species and transition metal ions", Free Radic. Biol, 30, pp.943~956 (2001)
- Synder R, "Christine C.H., An overview of benzene metabolism", Health Perspect, 104(6), pp.1165~1171 (1996)
- Zhang L, "Benzene metabolite, 1,2,4-benzentriol, induces micronuclei and oxidative DNA damage in human lymphocytes and HL-60 cells", Environ. Mol. Mutagen, 21, pp.339~348 (1993)
- Zhang L, "Detection of 1,2,4-benzentriol induced aneuploidy and microtubule disruption by fluorescence in situ hybridization and immunocytochemistry", Mutat, 320, pp.315~327 (1994), 315-327
- Sakurai M.H, "B-cell proliferaion activity of 대한예방한의학회지 제12권 제2호(2008년 8월) 58 peptic polysaccharide from a medicinal herb, the roots of Bupleurum falcatum L.and its stuctural requirement", Immunology, 97, pp.540~547 (1998)
- 강병수, "임상배합본초학", pp.494~497 (1994), 성보사
- 신민교, "원색임상본초학", 남산당, pp.313~314 (1986)
- 이상인, "본초학", 학림사, pp.198~199 (1986)
- 이상인, "한약임상응용", pp.74~76 (1982), 성보사
- Seto H, "tudies on chemical constituents of Bupleurumgenus.I. A. new triterpenoid glycosidefrom Bupleurum chinense DC", Agric. Biol. Chem, 50, pp.939~942 (1986), 939-942
- Guinea M.C, "Biologically active triterpenesaponins from Bupleurum Fruicosum", Planta medica, 60, pp.163~167 (1994)
- Kok L.D.S., "Activationof the anti-tumor effector cells byRadix bupleuri", Immnopharmacology, pp.79~87 (1995)
- Yen M.H, "Anti-inflammatory and hepatoprotectiveactivity of saikosaponin-f and theroot extract of Bupleurum kaoi", Fitoterapia65, 65, pp.409~417 (1994)
- Motoo Y, "Antitumor effectsof saikosaponins, baicalin and baicaleinon human hepatoma cell lines", Cancer Lett, 86, pp.91~95 (1994)
- Abe H, "Protective effect of saikosaponin-disolated from Bupleurum falcatum L. onCCl4-induced liver injury in the rat", Naunyn Schmiedebergs Arch Pharmacol, 320(3), pp.266~271 (1982)
- Stopper H, "Micronuclei as abiological endpoint for genotoxicity : aminireview. Toxicol", In vitro, 11, pp.661~667 (1997), aminireview.Toxicol.In vitro 11,661-667
- Sierens J, "Effect ofphatoestrogen and antioxidant supplementationon oxidative DNA damageassessed using the comet assay", Mutat, 485, pp.169~176 (2001)
- Fenech M, "Measurement ofmicronuclei in lymphocytes", Mutat, 147, pp.2936~ (1985)
- Fenech M, "The in vitro micronucleustechnique", Mutat, 455, pp.8195~2000 (2000), 8195
- Singh N.P, "Modifications of alkaline microgelelectrophoresis for sensitive detection ofDNA damage", Radiat. Biol, 66(1), pp.23~28 (1994)
- Wang H, "Quantifying cellularoxidative stress by dichlorofluorescein assayusing microplate reader", Free Radic. Biol.Med, 27, pp.612~616 (1999)
- Chiang L.C, "Cytotoxicity and antihepatitisB virus activities of saikosaponinsfrom Bupleurum species", PlantaMed, 69, pp.705~709 (2003)
- Matsumoto T, "Role of polymorphonuclear leucocytes andoxygen-derived free radicals in the formationofgastric lesions induced by HCl", ethanol, and a possible mechanism ofprotection by anti-ulcer polysaccharide.J. Pharm. Pharmacol, 1993, pp.45(6), 535~539
- Kun du T, "Role of polymorphonuclearleucocytes and oxygen-derived free radicalsin the formation of gastric lesions inducedby HCl", ethanol, and a possible mechanismof protection by anti-ulcer polysaccharide, 1993, pp.45(6), 535~539
- Yen M.H, "The hepatoprotective effect of Bupleurum kaoi, an endemic plant to Taiwan, against dimethylnitrosamine-induced hepatic fibrosis in rats", Biol Pharm. Bull, 28(3), pp.442~448 (2005)
- Lin C.C, "The pharmacological and pathological studies on Taiwan folk medicine(III) : The effects of bupleurum kaoi and cultivated bupleurum falcatum var. komarowi", Chin, 18, pp.105~112 (1990)
- Wang B.J., "Hepatoprotective and antioxidant effects of Bupleurum kaoi Liu(Chao et Chuang) extract and its fractions fractionated using supercritical CO2 on CCl4- induced liver damage", Food Chem. Toxicol, 42(4), pp.609~617 (2004)
- Hattori T, "Studies on antinephritic effects of plant components in rats(1). Effects of saikosaponins originaltype anti-GBM nephritis in rats and its mechanisms", Nippon Yakurigaku Zasshi, 97(1), pp.13~21 (1991)