线粒体DNA3243A>G突变糖尿病外周血单个核细胞生物力学变化特征
投稿时间:2017-08-15  修订日期:2017-09-20  点此下载全文
引用本文:耿新倩,张宜男,王从容.线粒体DNA3243A>G突变糖尿病外周血单个核细胞生物力学变化特征[J].医学研究杂志,2018,47(5):55-59
DOI: 10.11969/j.issn.1673-548X.2018.05.014
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作者单位E-mail
耿新倩 200233 上海交通大学附属第六人民医院内分泌代谢科、上海市糖尿病研究所、上海市糖尿病重点实验室、上海市糖尿病临床医学中心  
张宜男 200233 上海交通大学附属第六人民医院转化医学中心、上海市糖尿病重点实验室、上海交通大学附属第六人民医院代谢性疾病样本库  
王从容 200233 上海交通大学附属第六人民医院内分泌代谢科、上海市糖尿病研究所、上海市糖尿病重点实验室、上海市糖尿病临床医学中心 crwang@sjtu.edu.cn 
基金项目:上海市重大疾病临床生物样本实体库专业技术服务平台项目(15DZ2292100);上海交通大学多学科交叉项目培育(医工)基金资助项目(YG2016MS16);上海交通大学医学院转化医学创新基金资助项目——精准医学研究项目(15ZH4006)
中文摘要:目的 探索线粒体DNA3243A>G (mt.3243A>G)突变糖尿病患者和正常对照外周血单个核细胞(PBMC)表面形貌和力学性能差异。方法 采集5例mt.3243A>G突变糖尿病患者和5例年龄、性别匹配的正常对照外周血2ml,然后利用聚蔗糖-泛影葡胺(Ficoll-hypaque)密度梯度离心法分离出PBMC。应用原子力显微镜(AFM)对突变患者和正常对照的PBMC进行表面形貌和力学性能测量。结果 运用AFM测量和分析发现,在表面形貌方面,mt.3243A>G突变糖尿病患者的PBMC高度(0.73±0.24μm vs 2.49±1.17μm,P=0.011)低于正常对照组;但其表面粗糙度(Ra:161.8±33.2nm vs 66.4±16.3nm,P=0.000;Rq:202.2±40.9nm vs 85.4±17.1nm,P=0.000)高于正常对照组。在力学性能方面,mt.3243A>G突变糖尿病患者PBMC黏附力约比正常对照组高3倍(779.6±190.0pN vs 161.1±83.1pN,P=0.000)。与正常对照组相比,mt.3243A>G突变糖尿病患者PBMC杨氏模量(138.3±77.2kPa vs 421.4±140.0kPa,P<0.01)显著增加,病变细胞表面硬度增加。结论 本研究利用AFM从单细胞水平上揭示了mt.3243A>G突变糖尿病患者PBMC的表面形貌和力学性能变化,有助于加深对该疾病病理生理机制的理解。
中文关键词:mt.3243A>G突变  糖尿病  外周血单个核细胞  原子力显微镜
 
Biomechanical Characteristics of Peripheral Blood Mononuclear Cells in Mitochondrial Diabetes Caused by mt.3243A > G Mutation
Abstract:Objective To explore the surface morphological and biomechanical properties differences of peripheral blood mononuclear cells (PBMCs) between groups of patients with mitochondrial diabetes caused by mt.3243A>G mutation and healthy controls.Methods 2 milliliters blood were obtained from each subject of the mitochondrial diabetes group (n=5) and the control group (n=5). The PBMCs were separated from the blood using the standard Ficoll-Hypaque density-gradient centrifugation method and detected by atomic force microscope (AFM).Results The morphological analysis revealed that compared with control group, the PBMCs of diabetic patients tended to have a lower cell height (0.73±0.24μm vs 2.49±1.17μm,P=0.011) and a much rougher cell membrane (Ra:161.8±33.2nm vs 66.4±16.3nm,P=0.000;Rq:202.2±40.9nm vs 85.4±17.1nm,P=0.000). The adhesion force distribution was nearly three times higher in PBMCs of diabetic patients than that of the control group (779.6±190.0pN vs 161.1±83.1pN,P=0.000). The Young's modulus of PBMCs was significantly increased in diabetic patients (421.4±140.0kPa vs 138.3±77.2kPa,P<0.01), indicating that diabetic PBMCs were stiffer than control cells.Conclusion Our study demonstrated the surface morphological and biomechanical properties changes in mitochondrial diabetes caused by mt.3243A>G mutation at PBMCs level, which was beneficial to the better understanding of the pathophysiological mechanisms of mitochondrial diabetes associated with mt.3243A>G mutation.
keywords:mt.3243A>G mutation  Diabetes mellitus  Peripheral blood mononuclear cell  Atomic force microscope
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