硫化汞对灭活菌喂饲的秀丽隐杆线虫生长发育的影响
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摘要
目的探索线虫化学品毒性测试模型中喂饲用大肠杆菌(OP50)灭活的优化条件,确定钴60灭活OP50的最佳照射剂量。以秀丽隐杆线虫(简称线虫)的体长、体宽和咽泵震动频率为检测终点,研究硫化汞(HgS)对线虫幼虫生长发育的影响。方法采用钴60不同照射剂量(8、10、20和30 kGy)灭活的OP50(未浓缩和10∶1浓缩)及未处理的OP50分别喂饲L1期线虫,在0、24、48和72 h分别测量线虫的体长和体宽,以喂饲48 h对线虫体长和体宽无影响的钴60照射剂量为优选实验条件。在此实验条件下,线虫幼虫暴露于不同浓度的HgS(10、60和120 mg/ml)48 h,测定线虫体长、体宽和咽泵震动频率的变化。结果不同喂饲时间、不同照射剂量对线虫的生长发育影响不同。大于等于10 kGy的钴60照射剂量可将OP50完全灭活,未浓缩OP50经10 kGy的钴60灭活后喂饲线虫48 h,对其体长和体宽均无明显影响,故确定10 kGy为钴60的最佳照射剂量,且OP50不浓缩。在此优化条件下,L1期线虫暴露于60和120 mg/ml的HgS 48 h,线虫体长和体宽均减小;10 mg/ml的HgS对线虫生长发育没有影响。不同浓度的HgS对线虫的咽泵震动频率未见明显影响。结论在灭活菌喂饲的优化实验条件下,60 mg/ml及以上浓度的HgS对线虫的生长发育具有一定的抑制作用。
Objective To determine the optimal irradiation dose for ~(60)Co inactivation of Escherichia coli OP50 feeding Caenorhabditis elegans. Under the optimized irradiation condition, the effects of mercury sulfide(HgS) on the growth and development in L1 larvae were evaluated by detecting body length, body width and the pharyngeal pumping rate. Methods After the E. coli OP50(unconcentrated and 10∶1 concentrated) was inactivated by different doses(8, 10, 20, 30 kGy) of ~(60)Co, the body length and body width of C.elegans were observed at 0, 24, 48, 72 h, respectively. Based on the inactivation effect of OP50, and no significant influence on the body length and body width of C.elegans at 48 hours, the appropriate irradiation dose for inactication of OP50 was selected. With feeding of inactived E. coli OP50, the L1 larvae were treated with different concentrations of HgS(10, 60, 120 mg/ml) for 48 h, and the alterations of the body length, body width and the pharyngeal pumping rate were determined. Results The ~(60)Co irradiation dose of 10 kGy or more can completely inactivate E. coli OP50. The suitable irradiation condition is that unconcentrated E. coli OP50 inactivated by 10 kGy ~(60)Co. Fed with E. coli OP50 inactivated by 10 kGy ~(60)Co, the body length of C. elegans was shorter and the body width was narrow after exposed to HgS(60 and 120 mg/ml) for 48 h, compared to the solvent control. While the growth status of C. elegans exposed to HgS at low concentration(10 mg/ml) was not significantly changed. The test concentrations of HgS had no significant effect on the pharyngeal pumping rate of C. elegans. Conclusion Fed with inactivated E. coli OP50 by 10 kGy ~(60)Co, HgS at high concentration(≥60 mg/ml) inhibited the growth and development of C. elegans.
Objective To determine the optimal irradiation dose for ~(60)Co inactivation of Escherichia coli OP50 feeding Caenorhabditis elegans. Under the optimized irradiation condition, the effects of mercury sulfide(HgS) on the growth and development in L1 larvae were evaluated by detecting body length, body width and the pharyngeal pumping rate. Methods After the E. coli OP50(unconcentrated and 10∶1 concentrated) was inactivated by different doses(8, 10, 20, 30 kGy) of ~(60)Co, the body length and body width of C.elegans were observed at 0, 24, 48, 72 h, respectively. Based on the inactivation effect of OP50, and no significant influence on the body length and body width of C.elegans at 48 hours, the appropriate irradiation dose for inactication of OP50 was selected. With feeding of inactived E. coli OP50, the L1 larvae were treated with different concentrations of HgS(10, 60, 120 mg/ml) for 48 h, and the alterations of the body length, body width and the pharyngeal pumping rate were determined. Results The ~(60)Co irradiation dose of 10 kGy or more can completely inactivate E. coli OP50. The suitable irradiation condition is that unconcentrated E. coli OP50 inactivated by 10 kGy ~(60)Co. Fed with E. coli OP50 inactivated by 10 kGy ~(60)Co, the body length of C. elegans was shorter and the body width was narrow after exposed to HgS(60 and 120 mg/ml) for 48 h, compared to the solvent control. While the growth status of C. elegans exposed to HgS at low concentration(10 mg/ml) was not significantly changed. The test concentrations of HgS had no significant effect on the pharyngeal pumping rate of C. elegans. Conclusion Fed with inactivated E. coli OP50 by 10 kGy ~(60)Co, HgS at high concentration(≥60 mg/ml) inhibited the growth and development of C. elegans.
引文
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[2] 杨虹,彭芳,刘杰,等.朱砂安神丸、朱砂、硫化汞和氯化汞对大鼠肝脏细胞色素P450酶的影响[J].中国药理学与毒理学杂志,2012,26(2):231-236.
[3] 丁通,骆骄阳,韩旭,等.朱砂毒性的研究进展及配伍必要性分析[J].中国中药杂志,2016,41(24):4533-4540.
[4] 孙娜,田华,王蔚,等.重金属对秀丽隐杆线虫繁殖的影响[J].环境科学研究,2016,29(5):716-723.
[5] Hunt PR.The C.elegans model in toxicity testing[J].J Appl Toxicol,2017,37(1):50-59.
[6] Ma L,Zhao Y,Chen Y,et al.Caenorhabditis elegans as a model system for target identification and drug screening against neurodegenerative diseases[J].European J Pharmacol,2018,819:169-180.
[7] 张楠,李煜,高婷,等.以秀丽隐杆线虫作为人类疾病模型的研究进展[J].中国食品卫生杂志,2014,26(4):398-403.
[8] 高大文,高艳,张艳春,等.以不同方式灭活的大肠杆菌OP50饲喂对秀丽隐杆线虫生长发育的影响[J].军事医学,2013,37(3):215-218.
[9] Zhou X,Wang L,Sun X,et al.Cinnabar is not converted into methylmercury by human intestinal bacteria[J].J Ethnopharmacol,2011,135(1):110-115.
[10] 雷庆,黄敏,伍玲,等.辐照技术在检疫处理方面的应用概况[C].中国核学会2011年年会论文集,2011:61-64.
[11] 岳广智,杨立宏,徐宏山,等.钴60-γ射线辐照法对骨蜡中指示病毒灭活效果的验证[J].中国生物制品学杂志,2013,26(9):1318-1321.
[12] Yang J,Zhao Y,Wang YW,et al.Toxicity evaluation and translocation of carboxyl functionalized graphene in Caenorhabditis elegan[J].Toxicol Res,2015,4(6):1498-1510.
[13] 马娜娜,张楠,孔玲,等.硫柳汞对秀丽隐杆线虫神经行为及生长发育的影响[J].毒理学杂志,2017,31(2):88-93.
[14] 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.
[15] 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.