Honma Hiroyuki, Gross LouAnn, Windebank Anthony J
Department of Neurology, Cellular Neurobiology Laboratory, Mayo Clinic College of Medicine, 200 First Street SW, Rochester, MN 55905, USA.
Neurosci Lett. 2004 Jan 9;354(2):95-8. doi: 10.1016/j.neulet.2003.08.084.
The metabolic effects of hyperglycemia and hypoxia are important in the pathogenesis of diabetic neuropathy. We demonstrated apoptosis in dorsal root ganglion neurons in vitro by employing an oxygen-glucose deprivation model that uses dorsal root ganglia incubated in room air (pO2=150 torr) followed by hypoxic conditions (pO2=7.6 torr). Apoptosis was confirmed by demonstrating caspase-3 activation by immunocytochemistry. Immunocytochemistry and western blot analysis demonstrated an increase in activated p53, suggesting that DNA damage was occurring. Cell cycle disruption was examined by cyclin D1 expression. Neuronal death was associated with up-regulation of markers associated with DNA damage and aberrant entry into G1 of the cell cycle.
高血糖和缺氧的代谢效应在糖尿病神经病变的发病机制中具有重要作用。我们通过采用氧-葡萄糖剥夺模型在体外证明了背根神经节神经元的凋亡，该模型是将背根神经节置于室内空气（pO2 = 150托）中孵育，随后置于低氧条件（pO2 = 7.6托）下。通过免疫细胞化学证明半胱天冬酶-3激活来确认凋亡。免疫细胞化学和蛋白质印迹分析表明活化的p53增加，提示正在发生DNA损伤。通过细胞周期蛋白D1表达检测细胞周期紊乱。神经元死亡与DNA损伤相关标志物的上调以及异常进入细胞周期的G1期有关。
