HepG2细胞中线粒体形状的动态变化

Dynamic Change of Mitochondria Morphology in HepG2 Cells

目的:研究HepG2细胞中线粒体形状动态变化过程中的功能变化及其初步分子机制。方法:HepG2细胞经过HBSS缓冲液饥饿处理后,使用线粒体氧化磷酸化解偶联剂CCCP、脂肪酸受体GPR40/120激动剂GW9508、脂肪酸油酸OA和钙离子载体Ionomycin等4种不同药物处理,通过共聚焦显微镜观察和流式细胞分析的手段检测细胞中线粒体形状和功能发生的改变。然后,通过基因沉默Drp1,Mff或者Fis1蛋白,初步研究调控线粒体形状改变的分子机制。结果:经过CCCP和GW9508处理细胞中产生甜甜圈线粒体,而OA和Ionomycin处理产生球状线粒体。CCCP,OA和Ionomycin使线粒体去极化,CCCP、GW9508、OA或者Ionomycin单独处理在一定程度上影响细胞中活性氧化簇ROS。甜甜圈线粒体产生由Drp1介导,而球状线粒体形成依赖于Drp1和Mff。结论:线粒体的形态与其功能相互联系,Drp1和Mff蛋白对于细胞线粒体形状动态改变过程中形状的调整和适应具有很重要的作用。

Objective:To investigate the functional alteration and preliminary molecular mechanism of mitochondria morphology dynamic change in HepG2 cells.Method(s):After starved by HBSS buffer,HepG2 cells were treated with 4 kinds of different drugs including mitocho ndrial oxidative phosphorylation uncoupler CCCP,fatty acid receptor GPR40/GPR120 agonist GW9508,oleic acid OA and calcium ionophore Ionomycin,then mitochondria morphology and function were observed and measured by confocal microscope observation and flow cytometry analysis.The molecular mechanism of mitochondria morphology dynamic change was preliminarily studied by knock-down of dynamin-related protein 1(Drp1),mitochondrial fission factor(Mff)or fission mitochondrial 1(Fis1).Result(s):Cells had Donut mitochondria with mitochondrial oxidative phosphorylation uncoupler CCCP or fatty acid receptor GPR40/120 agonist GW9508 treatment while Ball mitochondria happened with OA or Iomomycin treatment.CCCP,OA and Ionomycin depolarized mitochondria,and CCCP,GW9508,OA or Ionomycin alone affected the reactive oxygen species(ROS)in cells to some extent.Donut mitochondria formation was mediated by Drp1,whereas Ball mitochondria formation was dependent on Drp1 and Mff.Conclusion(s):The morphology of mitochondria is related to its function,Drp1 and Mff play an important role in mitochondria morphology adjustment.