基于毒性效应通路荧光细胞传感模型的T-2毒素毒性筛查

doi:10.3969/j.issn.1673-1689.2021.12.005

首页 > 过刊浏览>2021年第40卷第12期 >35-43. DOI:10.3969/j.issn.1673-1689.2021.12.005

基于毒性效应通路荧光细胞传感模型的T-2毒素毒性筛查
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                        10.3969/j.issn.1673-1689.2021.12.005
                    
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T-2 Toxicity Screening Based on Fluorescence Cell Sensing Model via Toxic Effect Pathway

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为实现T-2毒素的可视化毒性筛查，利用T-2毒素毒性通路中AP-1的关键靶点应答元件TRE与红色荧光蛋白（mCherry）的启动子构建pcDNA3.1-TRE-mCherry荧光质粒，建立荧光HEK293细胞传感筛查模型；同时，为验证该细胞传感模型的适用性，对实际样品中残留的T-2毒素和T-2毒素的主要代谢产物HT-2毒素进行了毒性监测。结果表明，T-2毒素刺激模式细胞8 h时，细胞内的红色荧光强度达到最高值并趋于稳定。当T-2毒素质量浓度在1~25 ng/mL时，其与荧光强度呈线性相关，线性方程为y=1.149 38x+64.72，R²=0.969。根据剂量曲线得出T-2毒素的EC₅₀为16.27 ng/mL，检测限为0.691 ng/mL。该细胞传感模型用于样品加标实验，平均加标回收率为86.13%~126.20%，对HT-2毒素进行毒性筛查得到EC₅₀为27.65 ng/mL。

Abstract:

To realize the visual toxicity screening of T-2 toxin, the fluorescent plasmid pcDNA 3.1-TRE-mCherry was constructed using the key AP-1 target response element TREand red fluorescent protein (mCherry) in the toxicity pathway of T-2 toxin and the sensing screening model of fluorescent HEK293 cells was established. To verify the applicability of the cell sensing model, the toxicity detection of T-2 toxin residue in the actual samples and HT-2 toxin which was the main metabolite of T-2 toxin was conducted. The results showed that the intracellular red fluorescence intensity reached the maximum value and tended to be stable when the T-2 toxin stimulated the model cells for 8 h. When the concentration of T-2 toxin was in the range of 1~25 ng/mL, it was linearly correlated with fluorescence intensity, and the linear equation was y=1.149 38x+64.72, R2，R²= 0.969. According to the dose curve, the EC₅₀ of T-2 toxin was 16.27 ng/mL, and the detection limit was 0.691 ng/mL. The cell sensing model was used in the standard addition test. The average recovery of standard addition was 86.13%~126.20%, and the EC₅₀ of HT-2 toxin was 27.65 ng/mL after toxicity screening.

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孙嘉笛,高璐,张银志,孙秀兰.基于毒性效应通路荧光细胞传感模型的T-2毒素毒性筛查[J].食品与生物技术学报,2021,40(12):35-43.

SUN Jiadi, GAO Lu, ZHANG Yinzhi, SUN Xiulan. T-2 Toxicity Screening Based on Fluorescence Cell Sensing Model via Toxic Effect Pathway[J]. Journal of Food Science and Biotechnology,2021,40(12):35-43.

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在线发布日期: 2021-12-27
出版日期: 2021-12-25

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