不同诱导方式制备的大鼠肝匀浆代谢酶活性及冻储方式的比较

doi:10.3969/j.issn.1004-616x.2016.05.011

Abstract

Abstract: OBJECTIVE: To investigate the metabolic activities of rat liver S9 after using 7 different kinds of inducting agents and cryopreservation methods. METHODS: The S9 fraction was prepared from male Sprague-Dawley rats with phenobarbital,β-naphthoflavone or PCB induction,using different routes of administration,dosage and time by the way of combination or single inducing method. The S9 fractions were stored at 0,-20 and -40℃. Three indirect mutagens were selected and used to test for mutagenicity:2-amino-fluorene(2AF),acridine orange(AO) and cyclophosphamide (CP). RESULTS: Each group of liver homogenates showed positive results when 2AF,AO and CP were used as indirect mutagens. The activity of S9 from the combined induction method was higher. S9 could be preserved with glycerinum and stored at -20 or -40℃ without obvious loss of activity. CONCLUSION: Our data showed the following levels of S9 metabolic activities:PCB induced group=combination induced group >single induced group >non-induced group. The different routes of administration,dosage and time had little influence on the activity of S9. The S9 fraction retained good activities after storage in -20 and -40℃.

Key words: liver homogenates, combination induction, Ames test, enzyme protecting agent

CLC Number:

R73-3

SHAN Chun, HU Peili, ZHANG Luyong, LIU Ting, LIN Fei. Different induction and cryopreservation methods on metabolic activities of rat liver S9[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2016, 28(5): 388-392.

References

[1] 李寿祺.毒理学原理与方法[M].2版.四川:四川大学出版社,2003:402-404.
[2] 范玉明,张舒.毒理学安全性评价标准操作规程指南:下册[M].四川:电子科技大学出版社,2009:25-26.
[3] Garcia FS,Dominguez G,Pico JC.Alternatives in the induction and preparation of phenobarbital/naphthoflavone-induced S9 and their activation profiles[J].Mutagenesis,1999,14(3):323-326.
[4] 国家食品药品监督管理局.化妆品卫生规范[S].北京:中国标准出版社,2008:126-127.
[5] 王亚其,李宏霞,肖凯,等.两种诱导方法制备大鼠肝S9在两种遗传毒性试验中活性比较[J].现代预防医学,2006,33(4):457-459.
[6] Maron DM,Ames BN.Revised methods for the Salmonella mutagenicity test[J].Mutat Res,1983,113(3/4):173-215.
[7] 国家食品药品监督管理局.GB/T 16175-2008医用有机硅材料生物学评价试验方法[S].北京:中国标准出版社,2008:27.
[8] Atsushi H,Satoshi S,Shigeki S,et al.An improvement of the Ames test using a modified human liver S9 preparation[J].J Pharmacol Toxicol Methods,2002,46:169-172.
[9] Yang Z,Ding Y,Zhang Y,et al.Rapid purification of truncated Taq DNA polymerase Stoffel fragment by boiling lysis of bacterial expression cultures[J].Biotechnol Appl Biochem,2008,50(2):71-75.
[10] 黄幸纾,陈星若.环境化学物致突变、致畸、致癌试验方法[M].浙江:浙江科学技术出版社,1985:48-51.
[11] 阮丽荣,徐友梅,郑其岚,等.不同方法诱导大鼠肝S9对前致癌物活化能力的比较[J].河南医科大学学报,1988,23(3):201-203.
[12] 陈秀娟,黄俊明,李欣,等.溪黄草袋泡茶的急性毒性和遗传毒性研究[J].癌变·畸变·突变,2014,26(5):382-385.
[13] 何黎黎,杨立开,邓黎,等.S9代谢活化酶系统制备及蛋白含量的测定[J].西南民族大学学报,2010,36(2):243-245.
[14] Youngblom J.Extended stability of Taq DNA polymerase and T4 DNA ligase at various temperatures[J].Biotechniques,2003,34(2):264-268.

[1]	ZHOU Su, LI Rui, ZHU Hongyan, ZHANG Zhichao, FANG Jing, TANG Liming. An optical density method for determining bacterial concentrations in the Salmonella typhimurium reverse mutation assay [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2020, 32(1): 62-66.
[2]	ZHENG Kai, WANG Bingyu, XU Xinyun, GENG Hong, HUANG Haiyan, LIU Wei, LONG Dingxin. Mutagenicity of PM_2.5 samples from Shenzhen and Taiyuan [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2019, 31(6): 483-487.
[3]	CAO Yiyi, XI Jing, TANG Weifeng, YOU Xinyue, LIU Weiying, LUAN Yang. Genotoxicity evaluation of triclosan in vitro [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2018, 30(1): 71-75.
[4]	JIANG Yi, JIA Xiaohui, LI Peining, CHEN Xiujuan, LIU Donghong, LIU Xiangmei. Ames mutagenicity assay on five plastic surface chemicals in racetrack [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2017, 29(4): 309-312.
[5]	GAO Mei, CAO Chong, YING Yong, MA Hui, JIA Qingwen. Comparison of different inductionmethods on activity of rat liver microsomal enzymes [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2016, 28(4): 269-272.
[6]	YING Jian, HAO Binxiu, MENG Qingjia, CHEN Ran, WANG Liming, WANG Chunling. Acute toxicity and genotoxicity of crude water extracts from tea mistletoe Viscum Liquidambaricolum Hayata [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2016, 28(4): 306-309,324.
[7]	LUO Xueting, CAI Qijun, ZHAO Bo, CHEN Weimin, CHEN Xiao, ZHANG Xiaowen. Evaluation of acute and genetic toxicity of Zihe herbal solid beverage in mice [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2016, 28(4): 302-305.
[8]	TANG Jiao, ZHAO Min, YANG Ying, LI Qing, HUANG Junming, YANG Xingfen. Study on acute toxicity and mutagenicity of Panaxnotoginseng [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2016, 28(1): 66-68,72.
[9]	LIU Tiantian, SHEN Chunlin, LÜ Lulu, ZHANG Tianbao. Genotoxicity of PAMAM dendrimer (ethylenediamine core) generation 5.0 solution [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2015, 27(5): 378-382.
[10]	CHEN Xiu-juan，HUANG Jun-ming，LI Xin，CHEN Mei-fen，LI Qing，LIANG Yang-sheng，YI Zhi-mei. Study on the acute toxicity and genetic toxicity of rabdosia serra teabags [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2014, 26(5): 382-385.
[11]	WANG Chong，HOU Juan，HAN Jing，RUI Jing *. The safety and anti-mutagenicity of total terpene ketones from Swertia mussotii [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2014, 26(3): 221-224.
[12]	ZHANG Jian-jun，JIANG Ling，ZHANG Hai-jiao，CHAI Zhen-hai*. Effects of the ethylene oxide-sterilized Petri plate on Ames test [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2014, 26(3): 228-230.
[13]	HUO Jiao，ZHANG Meng-yun，ZHANG Li-shi，XU Pei-yu. The study of mutagenicity of 2-ethylhexyl acrylate [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2013, 25(6): 474-477.
[14]	ZHI Yuan，LIU Hai-bo，GENG Gui-ying，CUI Wen-ming，LIU Shan，XU Hai-bin*. Genetic toxicity of transgenic human α-lactalbumin powdered milk [J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2013, 25(2): 124-126.
[15]	QIN Xiu-jun;YUE Juan;LI Shu-fang;YIN Jing-jing;XU Wen-li;ZHENG Juan;LI Yun-lan;LI Qing-shan. Genotoxicity of di-phenyl-di- (2, 4 - chlorobenzohydroxamato) tin [J]. , 2012, 24(4): 305-308,.

Different induction and cryopreservation methods on metabolic activities of rat liver S9

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments