Nycodenz密度梯度离心法提取及纯化大鼠心肌线粒体  

Isolation and purification of rat myocardial mitochondria with Nycodenz density gradient centrifugation

作　　者：李科[1] 魏义勇[2] 刘云[3] 曹嵩[2] 刘兴奎[2] 王海英[2] 喻田[2] Li Ke Wei Yiyong Liu Yun Cao Song Liu Xingkui Wang Haiying Yu Tian(Department of Anesthesiology, Stomatological Hospital Affiliated to Zunyi Medical University, Zunyi Guizhou 563099, China Department of Anesthesiology, Affiliated Hospital of Zunyi Medical University, Zunyi Guizhou 563099, China Research Center for Medicine ＆ Biology, Zunyi Medical University, Zunyi Guizhou 563099, China)

机构地区：[1]遵义医学院附属口腔医院麻醉科,贵州遵义563099 [2]遵义医学院附属医院麻醉科,贵州遵义563099 [3]遵义医学院医学与生物学研究中心,贵州遵义563099

出　　处：《遵义医学院学报》2016年第5期525-528,共4页Journal of Zunyi Medical University

基　　金：卫生部公益性行业科研专项(NO:200802173);贵州省科技厅;遵义市科技局;遵义医学院联合基金资助项目(NO:黔科合J字LKZ[2013]31)

摘　　要：目的建立一种利用Nycodenz密度梯度离心法提取组织线粒体的方法。方法传统离心方法得到粗提线粒体后,在Nycodenz密度梯度介质中超高速离心得到纯化线粒体,进一步在经改良的线粒体分离介质中重复离心2次,得到高纯度的线粒体。利用透射电镜和Western Blot检测线粒体纯度。结果使用Nycodenz密度梯度离心法和线粒体分离介质提纯的线粒体量多、纯度较高(COXⅣ含量纯化组:粗提组=220%±26%vs.160%±34%,P<0.05),且电镜照片显示结构完整。结论使用Nycodenz密度梯度离心法可获得高纯度且结构较完整的线粒体。Objective To introduce a tissue mitochondria isolation and purification method with Nycodenz density gradient centrifugation. Methods Mitochondria obtained by traditional centrifugation method were subjected to super-centrifugation in different density Nycodenz layers before centrifugation twice in modified mitochondrial isolation media. Electron microscopy and Western Blot were employed to test the purity of mitochondria. Results Mitochondria obtained with Nycodenz density gradient centrifugation and our modified mitochondrial isolation media were more abundant and purer than that from the traditional method. Conclusion Nycodenz density gradient centrifugation is an optimal method to get pure and structural integrity mitochondria.

关 键 词：线粒体 密度梯度离心法 Nycodenz 大鼠 

分 类 号：R363[医药卫生—病理学]