克老素在脊髓损伤中的表达及相关性分析

Expression of anti-aging gene Klotho in rats with spinal cord injury

目的:观察大鼠脊髓损伤模型中克老素的表达变化,分析克老素的表达与神经损伤之间的相关性。方法:利用钳夹法制备大鼠脊髓损伤模型(SCI),并随机分为正常对照组(Sham)、损伤1周、2周、4周组。应用逆转录聚合酶链式反应(RT-PCR)、免疫荧光染色和蛋白印迹方法检测各实验组脊髓组织中克老素mRNA及蛋白的表达,利用ELISA法检测组织8-羟基脱氧鸟苷(8-OHdG)含量,并分析其相关性。结果:克老素在脊髓组织中表达,且损伤组脊髓组织中克老素mRNA及蛋白表达水平明显低于假手术组,且随病程延长而降低。ELISA检测结果显示,损伤组脊髓组织中8-OHdG含量明显高于假手术组,且随病程延长而增加,与克老素蛋白表达呈负相关关系。结论:克老素的表达变化与大鼠脊髓损伤之间呈负相关,提示克老素可能参与脊髓损伤后神经细胞退变的病理过程。

Objective： To investigate the expression of Klotho and the relationship between Klotho and spinal cord injury （SCI）. Methods： Spinal cord injury model was made by clamping method. Fifty SD rats were randomly divided into sham- operated group （Sham）, SCI 1 week group, SCI 2 weeks group and SCI 4 weeks group. The expression of Klotho mRNA and protein was observed by reverse transcription polymerase chain reaction （RT-PCR）, immunofluorescence and Western blotting. To detect the level of oxidative stress, 8-hydroxy-deoxyguanine （8-OHdG） was measured by ELISA, and the relationship between Klotho and 8-OHdG was analyzed. Results.. The expression of Klotho mRNA and protein decreased as the illness grew worse. The expression of 8-OHdG in the spinal cord injury group was significantly higher than that in the sham group, and the level was negatively correlated with the expression of Klotho. Conclusion： The expression of Klotho could be neganitively related to spinal cord injury of rats. It suggests that Klotho may be associated with neuronal degeneration after spinal cord injury.