细胞生物学实验教学改革实践探索--小鼠腹腔巨噬细胞的提取及NF-κB信号通路活化水平检测  

The Explorating Reform of Cell Biology Experiment Teaching--Extraction of Murine Peritoneal Macrophages and Detection of NF-κB Signaling Pathway Activation

作　　者：可月双 巴雪青 曾宪录[1] KE Yueshuang;BA Xueqing;ZENG Xianlu(Northeast Normal University,School of Life Sciences,Changchun 130024,China)

机构地区：[1]东北师范大学生命科学学院,长春130024

出　　处：《中国细胞生物学学报》2024年第6期1181-1190,共10页Chinese Journal of Cell Biology

基　　金：东北师范大学本科教学综合改革项目(第三期)资助的课题。

摘　　要：细胞生物学实验是细胞生物学课程教学的重要组成部分,对于帮助学生更好掌握相关理论知识,培养学生的实验操作能力、科研思维能力均具有重要作用。细胞信号转导是细胞生物学理论课内容的重要章节,目前在高校本科生细胞生物学实验教学中,鲜有关于细胞内信号通路转导检测和观察的实验。该团队根据自身的科研活动和教学经验累积设计了一个独立完整的综合性实验,包含4个相互联系的实验,意在探究小鼠腹腔巨噬细胞中NF-κB信号通路的活化情况,实验分别为:(1)小鼠腹腔巨噬细胞的提取及纯化;(2)炎性因子刺激下,RelA/p65在细胞内定位的观察;(3)小鼠腹腔巨噬细胞中炎症因子mRNA表达变化的检测;(4)小鼠腹腔巨噬细胞中炎症因子蛋白表达变化的检测。该综合性实验容易获得较纯的原代小鼠腹腔巨噬细胞,并且在提取的细胞中能够明显检测到细胞内信号通路的转导情况,实验结果易于观察。这样一个独立完整的综合性实验,不仅能够丰富和优化本科生细胞生物学实验教学的内容和模式,并且能够使学生从实验中真正深入地了解机体内炎症反应过程中信号通路的转导等相关知识,加深学生对于理论课程的理解。Cell Biology Experiment is an important part of Cell Biology courses and plays an important role in cultivating students’practice and scientific research thinking abilities.“Cell Signal Transduction”is the key and difficult chapter of Cell Biology course.The detection of intracellular signal transduction is an crucial experiment to the universities’s undergraduate students.Due to the complicated steps of signal transduction experiments and students are not familiar with the procedures,the experimental results are often not satisfactory,and it is difficult for students to observe significant phenomena.Therefore,in the current teaching of undergraduate Cell Biology Experiments in universities,there are few experiments on the detection and observation of intracellular signal transition.The purpose of this study is to investigate the activation of NF-κB signaling pathway in murine peritoneal macrophages.Four interrelated experiments were designed,which were:(1)extraction and purification of murine peritoneal macrophages;(2)observation of the localization of RelA/p65 in cells stimulated by inflammatory factors;(3)the mRNA expression of inflammatory factors in murine peritoneal macrophages were detected;(4)the protein expression of inflammatory factors in murine peritoneal macrophages were detected.This experiment is easy to isolate the murine peritoneal macrophages and observe the obvious relocation of RelA/p65.The independent and complete comprehensive experiment can enrich and optimize the content and model of undergraduate teaching of Cell Biology Experiment.Furthermore,the experiment can help students to truly understand the cell signaling pathways in the inflammatory response process.

关 键 词：细胞生物学实验 教学改革 小鼠腹腔巨噬细胞 NF-ΚB信号通路 细胞生物学实验教学 

分 类 号：G642.423[文化科学—高等教育学] Q2-4[文化科学—教育学]