基于微流控芯片-线虫模型的镉中毒和解毒的研究  

Research on cadmium poisoning and detoxification based on microfluidic chip-nematode modeling

作　　者：薛雪 邓立芬 陈柳生[1] 蔡自由[1] XUE Xue;DENG Li-fen;CHEN Liu-sheng;CAI Zi-you(Guangdong Food and Drug Vocational College,Guangzhou 510520,China)

机构地区：[1]广东食品药品职业学院,广州510520

出　　处：《药物分析杂志》2024年第6期1024-1030,共7页Chinese Journal of Pharmaceutical Analysis

基　　金：广州市科技计划(202102021276);广东省高校特色创新项目(2021KTSCX240);广东省中医药局项目(20211284);广东省中医药局(20231247)。

摘　　要：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0.25~15.0μg·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、5.0μg·mL^(-1)mL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 0.25-15.0μg·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 0.25,1.50,5.0,10.0,12.5,15.0μg·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.

关 键 词：秀丽隐杆线虫 镉中毒 维生素C 乙二胺四乙酸二钠钙 微流控芯片 解毒 药物筛选 

分 类 号：R917[医药卫生—药物分析学]