项目来源
上海市科技计划
项目主持人
陈红君
项目受资助机构
上海市杨浦区中心医院（同济大学附属杨浦医院）
立项年度
2019
立项时间
未公开
项目编号
19ZR1450600
研究期限
未知 / 未知
项目级别
省级
受资助金额
未知
学科
自然科学基金与基础重大重点研究
学科代码
未公开
基金类别
科技创新行动计划-基础研究领域项目
关键词
化学发光 ; DNA损伤 ; 信号放大 ; 多指标检测 ; 高通量筛选 ; DNA damage ; Chemiluminescence ; Signal amplification ; Multi-indicator detection ; High-throughput screening
参与者
未公开
参与机构
未公开
项目标书摘要：快速灵敏的评价环境污染物的遗传毒性并高通量筛选DNA保护药物对预防癌症、衰老、遗传疾病等具有重要意义。本研究在前期构建的基于磁性微球与酶联免疫化学发光检测DNA2种损伤(8-羟基脱氧鸟苷与DNA单链断裂(SSB))并高通量筛选天然DNA保护剂的技术基础上，引入了聚苯乙烯微球与双标记金纳米粒信号放大技术，优化了原来以DNA SSB为指标的高通量筛选模型的Z'因子，使其>0.8,同时构建了DNA甲基化,8-OHdG,DNA-苯并芘加合物以及DNA SSB 4种DNA损伤指标联合检测的技术，通过优化反应条件，排除了指标间检测的干扰，并考察了吸烟对4种指标产生的影响，阐述了吸烟致DNA各种损伤的关系，筛选了100多种天然产物效果，进一步在分子水平揭示了吸烟的遗传毒性，对于筛选出的6种高效，多效的DNA天然保护剂如半边莲，牛白藤，飞扬草，防风草，马尾松，山楂醇提物，在真实DNA与动物细胞水平进行了验证。本次优化的基于化学发光信号放大的DNA损伤多指标联合检测与天然保护药物高通量筛选体系既可应用于环境污染物的遗传毒性评价，又可应用于DNA保护药物的高通量高内涵筛选，方法经过了细胞及动物水平的验证，预计在新药研发方面具有广阔的应用前景。
Application Abstract: Rapid and sensitive evaluation of the genotoxicity of environmental pollutants and high-throughput screening of DNA protective natural products are of great significance for the prevention of cancer,aging and genetic diseases.In this study,on the basis of the previously constructed technology based on magnetic microspheres and enzyme-linked immunochemiluminescence to detect two kinds of DNA damage(8-hydroxydeoxyguanosine and DNA single strand break(SSB))and high-throughput screening of DNA protective natural products,the signal amplification technology of polystyrene microspheres and double labeled gold nanoparticles was introduced to optimize the Z'factor of the original high-throughput screening model with DNA SSB as the index,Z'>0.8.At the same time,The combined detection technology for 4 kinds of DNA damage(DNA methylation,8-OHdG,DNA benzopyrene adduct and DNA SSB)was developed and interference between the indexes was eliminated by optimizing the reaction conditions.Using this technology,the influence of smoking on the four kinds of DNA damage was inspected and the relationship between various DNA damages was expounded.Multi effective DNA protective natural products such as alcohol extracts of Lobelia chinensis L.,Hedyotis hedyotidea DC,Euphorbia hirta L.,Pastinaca sativa L.,PinusmassonianaLamb,and Crataegus pinnatifida were verified at the level of real DNA and animal cells.The optimized multi-indicator combined detection technology for DNA damage and the high-throughput screening system for natural products based on chemiluminescence signal amplification can be applied to both the genotoxicity evaluation of environmental pollutants and the high-throughput/high-content screening of DNA protective drugs.The method has been verified at the cellular and animal levels,and is expected to have broad application prospects in new drug research and development.
项目受资助省
上海市
