体内外染色体畸变实验检测朱砂的遗传毒性
|
摘要
目的:用仓鼠的骨髓细胞与CHL细胞经行体内外染色体畸变实验评价朱砂的遗传毒性。方法:体内染色体畸变实验用毛足属仓鼠给予朱砂混悬液(2.0,4.0,8.0 g·kg-1)连续灌胃5 d,处死前2 h腹腔注射秋水仙素。处死后取骨髓细胞制备染色体。油镜下观察每只动物骨髓细胞的有丝分裂指数和100个中期分裂相细胞的畸变类型。CHL细胞染色体畸变实验用朱砂浸出液终质量浓度为6.3,12.5,25.0 g·L-1作用于CHL细胞,培养24,48 h,终止培养前4 h加入秋水仙素。收获细胞,制备染色体,油镜下观察CHL细胞的200个中期分裂相细胞的畸变类型。结果:(1)与空白组比较,朱砂浸出液12.5,25.0 g·L-1给药组的CHL细胞染色体畸变率均升高(P<0.05,P<0.01),且有明显的量效关系。(2)仓鼠体内实验与朱砂血清给药CHL细胞的畸变率无显著性差异。结论:(1)在本实验中朱砂浸出液在12.5 g·L-1以上时能致体外CHL细胞染色体畸变,而朱砂混悬液在8.0 g·kg-1下仓鼠体内染色体畸变实验未出现阳性结果。(2)仓鼠体内染色体畸变实验可作为研究中药新药遗传毒性评价实验方法之一。
Objective: To evaluate the genotoxicity of Cinnabaris by using in vitro CHL cell and in vivo Hamster(Phodopus sungorus) chromosome aberration tests. Method: In vivo chromosome aberration test,hamsters(P. sungorus) were orally given cinnabar suspension at doses of 2. 0,4. 0,8. 0 g·kg-1 respectively for5 days. They were intraperitoneally injected with colchicine 2 hours before being put to death. Bone marrow cells were taken to prepare chromosome smears. Mitotic index of bone marrow cells and types of chromosome aberration in 100 metaphase cells per animal were observed under the microscope oil lens. In CHL cell chromosome aberration test,cinnabar extracts were used to react with cells,with the final concentration of 6. 3,12. 5,25. 0 g·L-1,and cultured for 24 or 48 h. Colchicine was added and cultured for 4 hours. The cells were collected to prepare chromosome smears. Types of chromosome aberration in 200 metaphase CHL cells per dose were observed under the microscope oil lens. Result:(1) Obvious chromosome aberrations were observed at doses of 12. 5,25. 0 g·L-1 in CHL cell chromosome aberration test(P < 0. 05, P < 0. 01).(2) Compared with the negative control,the chromosome aberration rates in both in vivo chromosome aberration test and in vitro CHL cell chromosome aberration test after administration showed no statistically significant difference,with no dose-effect relations. Conclusion:(1) Cinnabar can cause chromosome aberrations in vitro at the dose of 12. 5 g·L-1 in this test. However,there was no positive result in the cinnabar suspension chromosomal aberration test at the dose of 8. 0 g·kg-1.(2) Hamster(P. sungorus) in vivo chromosome aberration test can be used as an experimental method for evaluating the genotoxicity of new traditional Chinese medicines.
Objective: To evaluate the genotoxicity of Cinnabaris by using in vitro CHL cell and in vivo Hamster(Phodopus sungorus) chromosome aberration tests. Method: In vivo chromosome aberration test,hamsters(P. sungorus) were orally given cinnabar suspension at doses of 2. 0,4. 0,8. 0 g·kg-1 respectively for5 days. They were intraperitoneally injected with colchicine 2 hours before being put to death. Bone marrow cells were taken to prepare chromosome smears. Mitotic index of bone marrow cells and types of chromosome aberration in 100 metaphase cells per animal were observed under the microscope oil lens. In CHL cell chromosome aberration test,cinnabar extracts were used to react with cells,with the final concentration of 6. 3,12. 5,25. 0 g·L-1,and cultured for 24 or 48 h. Colchicine was added and cultured for 4 hours. The cells were collected to prepare chromosome smears. Types of chromosome aberration in 200 metaphase CHL cells per dose were observed under the microscope oil lens. Result:(1) Obvious chromosome aberrations were observed at doses of 12. 5,25. 0 g·L-1 in CHL cell chromosome aberration test(P < 0. 05, P < 0. 01).(2) Compared with the negative control,the chromosome aberration rates in both in vivo chromosome aberration test and in vitro CHL cell chromosome aberration test after administration showed no statistically significant difference,with no dose-effect relations. Conclusion:(1) Cinnabar can cause chromosome aberrations in vitro at the dose of 12. 5 g·L-1 in this test. However,there was no positive result in the cinnabar suspension chromosomal aberration test at the dose of 8. 0 g·kg-1.(2) Hamster(P. sungorus) in vivo chromosome aberration test can be used as an experimental method for evaluating the genotoxicity of new traditional Chinese medicines.
引文
[1]中国药典委员会.中华人民共和国药典.一部[M].北京:中国医药科技出版社,2015:137-138.
[2]黄琼,覃吉高,冉秀芬.皮疹腐蚀性消化道炎症朱砂中毒死亡3例临床分析[J].中国医药指南,2012,10(25):297-298.
[3]霍韬光,王海宇,林欣然,等.朱砂中汞的生物可接受性及其吸收与排泄[J].化学研究,2012,23(4):52-55,59.
[4]褚明宝,陈敏.中药不良反应及其防治对策[J].首都医药,2011,18(2):41.
[5]赵源,吴文斌,汤家铭.雄黄致体内外染色体畸变[J].中国实验方剂学杂志,2012,18(14):245-249.
[6]米金霞,吴文斌,谷颖敏,等.朱砂对孕大鼠母体及胚胎-胎仔发育的毒性作用[J].上海中医药大学学报,2011,25(5):79-82.
[7]张超超,吴文斌,汤家铭.微核试验和彗星试验检测朱砂的遗传毒性[J].中国实验方剂学杂志,2011,17(17):228-233.
[8]谷颖敏,李咏梅,姜昕,等.朱砂灌胃给药对大鼠生育力与早期胚胎发育毒性的研究[J].中国实验方剂学杂志,2011,17(9):226-231.
[9]吴文斌,汤家铭,祝晓雯,等.中药雄黄对孕大鼠母体和胚胎-胎仔发育的毒性[J].毒理学杂志,2009,23(5):380-383.
[10]陈海媚,谢晓芳,彭成.中药毒理学研究中体外细胞毒性的评价指标及检测方法[J].中国实验方剂学杂志,2017,23(22):202-210.
[11]黄珍祯,赵源,樊海艇,等.黑线毛足仓鼠实验动物化及生物学特性观察[J].实验动物与比较医学,2011,31(5):1-5.
[12]Prestone R J,Kean B J,Galloway S,et al.Mammalian in vivo cytogenetic assays:analysis of chromosome aberrations in bone marrow cells[J].Mutaes,1987,189(2):157-165.
[13]刘倩,马学盛.化浊行血汤含药血清血管内皮保护作用及其体内给药时效差异的研究[J].时珍国医国药,2009,20(10):2501-2502.
[14]韩佳寅,易艳,梁爱华,等.中药遗传毒性研究思路和方法[J].中国中药杂志,2015,40(14):2696-2700.
[15]罗雪婷,蔡绮君,赵博,等.紫荷植物固体饮料的急性毒性和遗传毒性试验研究[J].癌变·畸变·突变,2016,28(4):302-305.
[16]严晓莺,董菊,王明艳.遗传毒理学检测方法在中医药研究中的应用[J].湖北中医药大学学报,2011,13(2):62-64.
[17]鄢伟伦,李晓波.中药复方加水解酪蛋白肽粉功效及遗传毒性研究[J].食品与药品,2016,18(2):102-105.
[18]张翅,马悦,高慧敏,等.苦参化学成分研究进展[J].中国实验方剂学杂志,2014,20(4):205-214.
[19]唐仕欢,杨洪军.中医组方用药规律研究进展述评[J].中国实验方剂学杂志,2013,19(5):359-363.
[20]肖娟,王莹,王新宏,等.中药化学成分肠道菌群代谢的研究进展[J].中国实验方剂学杂志,2012,18(5):247-251.
[21]邹志远,宋有涛,杨雷,等.不同炮制方法对大黄的遗传毒性的减毒效果的研究[J].辽宁大学学报:自然科学版,2011,38(4):338-342.
[22]徐陆正,解清,闫赖赖,等.中成药可溶性汞形态分析方法及样品结果测定[J].实验技术与管理,2011,28(12):40-43.
[23]王金华,叶祖光.安宫牛黄丸研究现状[J].中国中药杂志,2004,9(2):119-122.
[2]黄琼,覃吉高,冉秀芬.皮疹腐蚀性消化道炎症朱砂中毒死亡3例临床分析[J].中国医药指南,2012,10(25):297-298.
[3]霍韬光,王海宇,林欣然,等.朱砂中汞的生物可接受性及其吸收与排泄[J].化学研究,2012,23(4):52-55,59.
[4]褚明宝,陈敏.中药不良反应及其防治对策[J].首都医药,2011,18(2):41.
[5]赵源,吴文斌,汤家铭.雄黄致体内外染色体畸变[J].中国实验方剂学杂志,2012,18(14):245-249.
[6]米金霞,吴文斌,谷颖敏,等.朱砂对孕大鼠母体及胚胎-胎仔发育的毒性作用[J].上海中医药大学学报,2011,25(5):79-82.
[7]张超超,吴文斌,汤家铭.微核试验和彗星试验检测朱砂的遗传毒性[J].中国实验方剂学杂志,2011,17(17):228-233.
[8]谷颖敏,李咏梅,姜昕,等.朱砂灌胃给药对大鼠生育力与早期胚胎发育毒性的研究[J].中国实验方剂学杂志,2011,17(9):226-231.
[9]吴文斌,汤家铭,祝晓雯,等.中药雄黄对孕大鼠母体和胚胎-胎仔发育的毒性[J].毒理学杂志,2009,23(5):380-383.
[10]陈海媚,谢晓芳,彭成.中药毒理学研究中体外细胞毒性的评价指标及检测方法[J].中国实验方剂学杂志,2017,23(22):202-210.
[11]黄珍祯,赵源,樊海艇,等.黑线毛足仓鼠实验动物化及生物学特性观察[J].实验动物与比较医学,2011,31(5):1-5.
[12]Prestone R J,Kean B J,Galloway S,et al.Mammalian in vivo cytogenetic assays:analysis of chromosome aberrations in bone marrow cells[J].Mutaes,1987,189(2):157-165.
[13]刘倩,马学盛.化浊行血汤含药血清血管内皮保护作用及其体内给药时效差异的研究[J].时珍国医国药,2009,20(10):2501-2502.
[14]韩佳寅,易艳,梁爱华,等.中药遗传毒性研究思路和方法[J].中国中药杂志,2015,40(14):2696-2700.
[15]罗雪婷,蔡绮君,赵博,等.紫荷植物固体饮料的急性毒性和遗传毒性试验研究[J].癌变·畸变·突变,2016,28(4):302-305.
[16]严晓莺,董菊,王明艳.遗传毒理学检测方法在中医药研究中的应用[J].湖北中医药大学学报,2011,13(2):62-64.
[17]鄢伟伦,李晓波.中药复方加水解酪蛋白肽粉功效及遗传毒性研究[J].食品与药品,2016,18(2):102-105.
[18]张翅,马悦,高慧敏,等.苦参化学成分研究进展[J].中国实验方剂学杂志,2014,20(4):205-214.
[19]唐仕欢,杨洪军.中医组方用药规律研究进展述评[J].中国实验方剂学杂志,2013,19(5):359-363.
[20]肖娟,王莹,王新宏,等.中药化学成分肠道菌群代谢的研究进展[J].中国实验方剂学杂志,2012,18(5):247-251.
[21]邹志远,宋有涛,杨雷,等.不同炮制方法对大黄的遗传毒性的减毒效果的研究[J].辽宁大学学报:自然科学版,2011,38(4):338-342.
[22]徐陆正,解清,闫赖赖,等.中成药可溶性汞形态分析方法及样品结果测定[J].实验技术与管理,2011,28(12):40-43.
[23]王金华,叶祖光.安宫牛黄丸研究现状[J].中国中药杂志,2004,9(2):119-122.