目的: 以秀丽隐杆线虫为实验对象研究金属镉的毒性以及解毒药物的筛选。方法: 以微流控芯片技术作为药物筛选平台,采用96孔板暴露预试验,以同期化后L4期的线虫,设置K-medium空白组,采用环境相关浓度为0.25~15.0 μg·mL-1的重金属镉设置暴露组,采用维生素C和乙二胺四乙酸二钠钙分别拮抗镉作为解毒组。每个孔板中加入线虫(10±2)条,在显微镜下观察记录线虫数量,观察线虫阴门结构的变化,实验数据均以origin 2019b软件进行操作处理,结合所得结果对数据进行分析。结果: 通过与空白组相比,24 h后暴露组线虫在0.25、1.50、5.0、10.0、12.5、5.0 μg·mL-1mL浓度下的致死率分别为0%、1.67%、4.76%、67.46%、100%、100%;线虫阴门结构有明显畸形变化,出现轻微凸起到严重凸起,甚至有瘤状物凸起,最后破裂。解毒组致死率与暴露组相比均有不同程度的减少。结论: 通过与空白对照组相比,随着镉暴露浓度和暴露时间的增加,镉元素对线虫的存活具有抑制作用,对线虫阴门结构有不同程度的损害。解毒组实验表明维生素C对金属镉的解毒作用较小;乙二胺四乙酸二钠钙与镉络合后可在较大程度上延长线虫的存活时间。
Objective: To study the toxicity of heavy metal cadmium and screen detoxification drugs base on Caenorhabditis elegans. Methods: The microfluidics chip technology was used as the drug screening platform. The 96-well plate exposure pretest was used, and K-medium blank group was set up. The exposure group consisted of nematodes exposed to heavy metal cadmium at the environmental relevant concentration of 0.25-15.0 μg·mL after homogenization. Vitamin C and calcium disodium ethylenediamine tetraacetate were selected as detoxification group. (10±2) nematodes were added into each orifice plate. The number of nematodes was observed and recorded under a microscope, and paid attention to the changes in the structure of the vulva. The experimental data were processed and analyzed by origin 2019b software. Results: Compared with blank group, the fatality rates of C. elegans in exposed group were respectively 0%, 1.67%, 4.76%, 67.46%, 100% and 100% at 0.25, 1.50, 5.0, 10.0, 12.5, 15.0 μg·mL-1 concentration, compared with blank group. The structure of the nematode vulva was deformed, from slight protrusion to severe protrusion, and even a tuberous protrusion, and finally ruptured. The mortality of detoxification group was reduced to different degrees compared with exposure group. Conclusion: Compared with blank control group, with the increase of cadmium exposure concentration and exposure time, cadmium can inhibit the survival of C. elegans, and damage the structure of C. elegans vulva to varying degrees. The results of detoxification group showed that vitamin C had little detoxification effect on cadmium. The complexation of calcium disodium ethylenediamine tetraacetate with cadmium could prolong the survival time of C. elegans to a large extent.
[1] 张晓华,肖雄斌. 镉毒性作用机制及临床防治进展[J]. 实用预防医学,2012,19(11):1761
ZHANG XH, XIAO XB. Mechanisms and clinical prevention and treatment progress of Cadmium poisoning[J]. Pract Prevent Med, 2012, 19(11):1761
[2] 张汉文,林家进,黄月娥,等. 基于秀丽线虫的5种重金属生物毒性评价[J]. 环境与健康杂志,2018,35(10):890
ZHANG HW, LIN JJ, HUANG YE, et al. Biotoxicity of five heavy metals to Caenorhabditis elegans[J]. J Environ Health, 2018, 35(10):890
[3] 丁通,骆骄阳,杨世海,等.天然药物防治镉中毒的现代研究进展[J]. 中国中药杂志,2018,43(10):2006
DING T, LUO JY, YANG SH, et al. Recent research progress on natural medicines in treatment of Cadmium toxicit[J]. China J Chin Mater Med, 2018, 43(10):2006
[4] 王晶晶,许安. 二氧化钛纳米颗粒的沉降过程促进重金属对秀丽隐杆线虫的生物积累与毒性效应[C]//中国化学会环境化学专业委员会. 第二届全国环境纳米技术及生物效应学术研讨会会议手册. 2018:102
WANG JJ, XU A. The sedimentation process of titanium dioxide nanoparticles promotes the bioaccumulation and toxic effects of heavy metals on Caenorhabditis elegans[C]//Environmental Chemistry Professional Committee of the Chinese Chemical Society. Handbook of the Second National Symposium on Environmental Nanotechnology and Biological Effects. 2018:102
[5] 张守申. 二氧化钛纳米与重金属Cd离子相互作用降低Cd2+对秀丽隐杆线虫的毒性[D]. 济南:山东大学,2014
ZHANG SS. Absorption on TiO2 Nanoparticles Reduced the Toxicity of Cd2+ to Caenorhabditis elegans[D]. Jinan:Shandong University, 2014
[6] 刁书永,张立志,袁慧. 镉中毒机理研究进展[J]. 动物医学进展,2005(5):49
DIAO SY, ZHANG LZ, YUAN H. Progress in the toxicity mechanism of cadmium[J]. Progr Veter Med, 2005(5):49
[7] WU L, AI Y, XIE R, et al. Organoids/organs-on-a-chip:new frontiers of intestinal pathophysiological model [J]. Lab Chip, 2023, 23(5):1192
[8] HUANG D, WANG J, NIE M, et al. Pollen-inspired adhesive multilobe microparticles from microfluidies for intestinal drug delivery[J]. Adv Mater, 2023, 35(8):e2301192
[9] 艾永建,何梦崎,梁琼麟. 基于微流控技术的药物分析新进展[J]. 药物分析杂志, 2023,43(10):1631
AI YJ, HE MQ, LIANG QL. Recent advances in microfluidics-based pharmaceutical analysis[J]. Chin J Pharm Anal, 2023, 43(10):1631
[10] YANG J, CHEN Z, CHING P, et al. An integrated microfluidic platform for evaluating in vivo antimicrobial activity of natural compounds using a whole-animal infection model[J]. Lab Chip, 2013, 13(17):3373
[11] 尹方超,温慧,朱国丽,等.基于微流控芯片技术的秀丽隐杆线虫研究进展[J]. 色谱,2016,34(11):1031
YIN FC, WEN H, ZHU GL, et al. Advances of Caenorhabditis elegans research on microfluidic device[J]. Chin J Chromatogr, 2016, 34(11):1031
[12] 李偶连,刘翠,陈缵光,等.微流控芯片技术及在药物分析中的应用研究进展[J]. 分析科学学报,2008(4):473
LI OL, LIU C, CHEN ZG, et al. The progress of microfluidic chip and its application in pharmaceutical analysis[J]. J Anal Sci, 2008(4):473
[13] 吴佳颖,李栋基,陈翔,等.基于微流控芯片技术的秀丽隐杆线虫药物筛选体系研究进展[J]. 药物分析杂志,2021,41(6):929
WU JY, LI DJ, CHEN X, et al. Advances in drug screening of C. elegans based on microfluidic chip techniques[J]. Chin J Pharm Anal, 2021, 41(6):929
[14] 赵晴,蒋湉湉. 秀丽隐杆线虫研究综述[J]. 安徽农业科学,2010,38(19):10092
ZHAO Q, JIANG TT. Overview of the Nematode Caenorhabditis elegans [J]. J Anhui Agr Sci, 2010, 38(19):10092
[15] 翟畅,叶波平. 秀丽隐杆线虫与药物筛选[J]. 药物生物技术,2017,24(5):464
ZHAI C, YE BP. Caenorhabditis elegans in drug screening[J]. Pharm Biotechnol, 2017, 24(5):464
[16] 魏召艺. 两种药物对秀丽隐杆线虫生命周期的影响研究[D]. 郑州:河南大学,2019
WEI ZY. Effect of Two Drugs on Caenorhabditis elegans Lifespan[D]. Zhengzhou:Henan University, 2019
[17] 李栋基,吴佳颖,李闯,等.用于秀丽隐杆线虫毒理学实验的类桥式微流控芯片平台[J]. 药物分析杂志,2020,40(7):1160
LI DJ, WU JY, LI C, et al. An bridge-like microfluidic chip based platform for toxicity testing of Caenorhabditis elegans[J]. Chin J Pharm Anal, 2020,40(7):1160
