Diminished superoxide generation is associated with respiratory chain dysfunction and changes in the mitochondrial proteome of sensory neurons from diabetic rats.
Diminished superoxide generation is associated with respiratory chain dysfunction and changes in the mitochondrial proteome of sensory neurons from diabetic rats.
复制标题
DOI：
10.2337/db10-0818
复制
发表时间：
2011-01
期刊：
影响因子：
7.7
通讯作者：
Fernyhough P
中科院分区：
文献类型：
--
作者：
Akude E;Zherebitskaya E;Chowdhury SK;Smith DR;Dobrowsky RT;Fernyhough P
线粒体功能受损被认为在糖尿病感觉神经病变的病因学中起作用。我们测试了一种假设，即1型糖尿病感觉神经元轴突中的线粒体功能障碍是由于呼吸链的异常活动和线粒体蛋白质组的改变所致。用细胞培养中氨基酸稳定同位素标记的蛋白质组学分析(SILAC)测定了对照组、22周龄链脲佐菌素(STZ)糖尿病大鼠和胰岛素治疗的糖尿病大鼠线粒体中蛋白质的表达。测定背根神经节线粒体的耗氧率和复合体活性。对培养的感觉神经元轴突的荧光成像确定了糖尿病对线粒体极化状态、氧化应激和线粒体基质特异性活性氧物种(ROS)的影响。与线粒体功能障碍、氧化磷酸化、泛醌生物合成和柠檬酸循环相关的蛋白质在糖尿病样本中下调。例如，细胞色素c氧化酶亚基IV(COX IV，一种复合IV蛋白)和NADH脱氢酶Fe-S蛋白3(NDUFS3，一种复合I蛋白)在糖尿病患者中分别减少了29%和36%(P<；0.05)。呼吸作用和线粒体复合体活性与对照相比显著降低15%~32%。糖尿病神经元的轴突表现出氧化应激和线粒体去极化，这是对寡霉素诱导的线粒体膜超极化的一种异常适应，但与对照组相比，线粒体内超氧化物水平降低。线粒体功能异常与线粒体蛋白下调有关，与糖尿病患者腰椎背根神经节的呼吸链成分有关。呼吸链活性的降低与线粒体基质中超氧化物生成的减少有关，与糖尿病神经元轴突的氧化应激无关。参与多元醇途径活动的其他途径似乎有助于在高糖浓度下糖尿病神经元轴突中ROS的升高。
Impairments in mitochondrial function have been proposed to play a role in the etiology of diabetic sensory neuropathy. We tested the hypothesis that mitochondrial dysfunction in axons of sensory neurons in type 1 diabetes is due to abnormal activity of the respiratory chain and an altered mitochondrial proteome. Proteomic analysis using stable isotope labeling with amino acids in cell culture (SILAC) determined expression of proteins in mitochondria from dorsal root ganglia (DRG) of control, 22-week-old streptozotocin (STZ)-diabetic rats, and diabetic rats treated with insulin. Rates of oxygen consumption and complex activities in mitochondria from DRG were measured. Fluorescence imaging of axons of cultured sensory neurons determined the effect of diabetes on mitochondrial polarization status, oxidative stress, and mitochondrial matrix-specific reactive oxygen species (ROS). Proteins associated with mitochondrial dysfunction, oxidative phosphorylation, ubiquinone biosynthesis, and the citric acid cycle were downregulated in diabetic samples. For example, cytochrome c oxidase subunit IV (COX IV; a complex IV protein) and NADH dehydrogenase Fe-S protein 3 (NDUFS3; a complex I protein) were reduced by 29 and 36% (P < 0.05), respectively, in diabetes and confirmed previous Western blot studies. Respiration and mitochondrial complex activity was significantly decreased by 15 to 32% compared with control. The axons of diabetic neurons exhibited oxidative stress and depolarized mitochondria, an aberrant adaption to oligomycin-induced mitochondrial membrane hyperpolarization, but reduced levels of intramitochondrial superoxide compared with control. Abnormal mitochondrial function correlated with a downregulation of mitochondrial proteins, with components of the respiratory chain targeted in lumbar DRG in diabetes. The reduced activity of the respiratory chain was associated with diminished superoxide generation within the mitochondrial matrix and did not contribute to oxidative stress in axons of diabetic neurons. Alternative pathways involving polyol pathway activity appear to contribute to raised ROS in axons of diabetic neurons under high glucose concentration.
登录
查看更多内容
DOI：
10.1073/pnas.1032913100
发表时间：
2003-07-08
影响因子：
11.1
作者：
Patti, ME;Butte, AJ;Mandarino, LJ
通讯作者：
Mandarino, LJ
影响因子：
2.9
作者：
Gardiner, Natalie J.;Wang, Zuocheng;Fernyhough, Paul
通讯作者：
Fernyhough, Paul
影响因子：
14.8
作者：
Frezza, Christian;Cipolat, Sara;Scorrano, Luca
通讯作者：
Scorrano, Luca
影响因子：
2
作者：
Obrosova, I;Faller, A;Williamson, JR
通讯作者：
Williamson, JR
DOI：
10.1152/ajpendo.00323.2005
发表时间：
2006-06-01
影响因子：
5.1
作者：
Ola, M. Shamsul;Berkich, Deborah A.;LaNoue, Kathryn F.
通讯作者：
LaNoue, Kathryn F.