Section of Nephrology, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA.
Department of Molecular & Cell Biology, Baylor College of Medicine, Houston, Texas, USA.
Kidney Int. 2017 Nov;92(5):1282-1287. doi: 10.1016/j.kint.2017.05.015. Epub 2017 Jul 26.
While increased mitochondrial reactive oxygen species have been commonly implicated in a variety of disease states, their in vivo role in the pathogenesis of diabetic nephropathy remains controversial. Using a two-photon imaging approach with a genetically encoded redox biosensor, we monitored mitochondrial redox state in the kidneys of experimental models of diabetes in real-time in vivo. Diabetic (db/db) mice that express a redox-sensitive Green Fluorescent Protein biosensor (roGFP) specifically in the mitochondrial matrix (db/dbmt-roGFP) were generated, allowing dynamic monitoring of redox changes in the kidneys. These db/dbmt-roGFP mice exhibited a marked increase in mitochondrial reactive oxygen species in the kidneys. Yeast NADH-dehydrogenase, a mammalian Complex I homolog, was ectopically expressed in cultured podocytes, and this forced expression in roGFP-expressing podocytes prevented high glucose-induced increases in mitochondrial reactive oxygen species. Thus, in vivo monitoring of mitochondrial roGFP in diabetic mice confirms increased production of mitochondrial reactive oxygen species in the kidneys.
虽然线粒体活性氧物质的增加通常与多种疾病状态有关，但它们在糖尿病肾病发病机制中的体内作用仍存在争议。本研究使用具有遗传编码氧化还原生物传感器的双光子成像方法，实时监测糖尿病实验模型肾脏中的线粒体氧化还原状态。生成了在基质中特异性表达氧化还原敏感型绿色荧光蛋白生物传感器（roGFP）的糖尿病（db/db）小鼠（db/dbmt-roGFP），从而可以动态监测肾脏中的氧化还原变化。这些 db/dbmt-roGFP 小鼠的肾脏中线粒体活性氧物质明显增加。在 roGFP 表达的足细胞中异位表达酵母烟酰胺腺嘌呤二核苷酸脱氢酶（一种哺乳动物复合物 I 同源物）可防止高葡萄糖诱导的线粒体活性氧物质增加。因此，糖尿病小鼠中 roGFP 标记的线粒体的体内监测证实了肾脏中线粒体活性氧物质产生的增加。
