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目的:探讨DJ-1基因L10P突变对细胞线粒体功能的影响。方法:应用流式细胞仪、分光光度计、电镜等技术对稳定表达空载体、野生型DJ-1蛋白、L10P突变型DJ-1蛋白的HEK293单克隆细胞株的生长活力、活性氧、膜电位、线粒体复合体酶I活性及线粒体形态进行分析。结果:与空载体组相比较,L10P突变型DJ-1组细胞内活性氧增高,细胞活力、膜电位、线粒体复合体I活性降低,线粒体含量减少(P<0.05),出现线粒体肿胀,甚至线粒体空泡变性,特别是在鱼藤酮的诱导下更明显;野生型DJ-1组细胞内活性氧均较低,细胞活力、膜电位、线粒体复合体I活性均较高(P<0.05),特别是在鱼藤酮的诱导下更明显。结论:L10P突变后使野生型DJ-1蛋白的抗氧化应激能力丧失,并对线粒体的正常功能存在影响。
Objective: To investigate the effect of L10P mutation of DJ-1 gene on mitochondrial function. Methods: The viability, reactive oxygen species and membrane potential of HEK293 cells stably expressing empty vector, wild type DJ-1 protein and L10P mutant DJ-1 protein were determined by flow cytometry, spectrophotometer and electron microscope. , Mitochondrial enzyme I activity and mitochondrial morphology were analyzed. Results: Compared with the control group, the level of reactive oxygen species (ROS) in the L10P mutant DJ-1 group was increased, the cell viability, membrane potential, mitochondrial I activity and mitochondria decreased (P <0.05) Vacuolar degeneration, especially under the induction of rotenone, the activity of ROS in wild-type DJ-1 group was lower, and cell viability, membrane potential and mitochondrial complex I activity were higher (P <0.05), especially More obvious under the induction of rotenone. CONCLUSION: L10P mutation causes the loss of antioxidant capacity of wild type DJ-1 protein and affects the normal function of mitochondria.