高内涵模型检测细胞焦亡用于抗炎药物筛选的实验教学设计  

Teaching design of the experiment“screening for anti-inflammatory agents by detection of pyroptosis based on a high content screening model”

作　　者：阮班锋 陈刘赠 吴克[1] 张敏[1] RUAN Banfeng;CHEN Liuzeng;WU Ke;ZHANG Min(School of Biology,Food and Environment,Hefei University,Hefei 230601)

机构地区：[1]合肥学院生物食品与环境学院,合肥230601

出　　处：《生物学杂志》2023年第4期127-130,共4页Journal of Biology

基　　金：国家自然科学基金青年基金项目(22207028);安徽省自然科学基金项目(2108085MH318);合肥学院“生物与医药”校级重点学科项目(2023xk05)。

摘　　要：介绍为国家级大学生创新创业训练项目设计的采用高内涵模型检测细胞焦亡用于筛选抗炎药物实验教学实施结果。该实验利用NLRP3炎症小体活化导致细胞焦亡的理论,使用高内涵对焦亡细胞进行计数,以确定化合物对NLRP3炎症小体活化抑制情况,进而评价化合物抗炎活性强弱。实验结果表明:NLRP3炎症小体活化时细胞会发生焦亡,PI能够对焦亡的细胞染红色荧光,Hoechst 33342能够对所有细胞染蓝色荧光。加入待筛选的化合物后,利用高内涵对不同荧光下的细胞进行计数,通过二者比值确定化合物的NLRP3炎性小体活化抑制活性,从而用于抗炎药物筛选。实验将PI/Hoechst 33342染色与高内涵结合使用,有效解决了经费不足和大型仪器设备缺乏等问题,实验成本较低,结果直观便捷,能够让学生深入、系统地了解细胞焦亡的实验原理和检测方法,让参加大学生创新创业项目的学生在掌握细胞生物学基本实验技能的同时进一步了解新药研发的基本流程,有效培养学生创新创业意识与科学素养。The teaching design of the experiment“screening for anti-inflammatory agents by detection of pyroptosis based on a high content screening model”was mainly introduced for the national college student innovation and entrepreneurship training program.Based on the theory of NLRP3 inflammasome activation leading to cell pyroptosis,the high content screening model was used to count the pyroptosis cells to determine the anti-inflammatory activity of the tested compound.The experimental results indicated that when NLRP3 inflammasomes were activated,cells underwent pyroptosis.PI could stain red fluorescence on pyroptosis cells,while the Hoechst 33342 could stain blue fluorescence on all cells.High content screening was used to count the cells with different fluorescence,the ratio was then used to determine the anti-inflammatory activity of the tested compound.The experiment combined the PI/Hoechst 33342 staining with high content screening,which could solve the problems of insufficient teaching funds,poor experimental conditions and so on.The experiment was convenient with low cost,the experimental results were intuitive and students could deeply,systematically and comprehensively understand the characteristics of the principle of pyroptosis.Thus,students who participated in the related innovation and entrepreneurship project of college students could further understand the basic process of new drug research and development while mastered the basic experimental skills of cell biology,effectively cultivated students'awareness of innovation and entrepreneurship and scientific literacy.

关 键 词：细胞焦亡 实验教学 高内涵 NLRP3炎性小体抑制剂 抗炎 

分 类 号：G642.1[文化科学—高等教育学] Q255[文化科学—教育学]