大鼠脊髓损伤过程中细胞周期蛋白H的表达变化

目的 观察细胞周期蛋白H(Cyclin H)在急性脊髓损伤后的表达变化及定位情况,探讨其在脊髓损伤与修复过程中的生物学功能.方法 取56只成年SD大鼠,随机分8组:正常对照组、T9撞击伤6 h、12 h、1 d、3 d、5 d、7 d、14 d 7组,每组7只.采用Western blot测定损伤后各时间段Cyclin H蛋白水平在脊髓中的表达变化,采用免疫组织化学方法检测Cyclin H在正常及损伤后脊髓中的分布和定位,在此基础上探讨Cyclin H在脊髓损伤后的病理生理意义.结果 Western blot结果表明创伤性脊髓损伤后脊髓中Cyclin H表达呈现逐渐升高再下降的趋势,3 d达到高峰.免疫组织化学表明Cyclin H在正常脊髓中表达,损伤后3 d,Cyclin H在脊髓白质中的表达明显增强,在灰质中的表达也有一定程度的增强.免疫荧光双标记表明Cyclin H与神经元标记物NeuN、小胶质细胞标记物CD11b有明显共定位,与星形胶质细胞标记物GFAP有少量共定位.结论 脊髓损伤后Cyclin H蛋白水平有明显时相改变,且与神经元、小胶质细胞和星形胶质细胞有共定位,提示Cyclin H在脊髓损伤后可能发挥了作用.

Expression of cyclin H following spinal cord contusion in rats

Objective To investigate the expression and distribution of cyclin H in rat models of spinal cord injury (SCI) .Methods Fifty-six adult Sprague-Dawley (SD) rats were randomized into 8even groups (one normal group and 7 SCI groups) .Contusion was created at T9 in the SD rats in the SCI groups.The rats in the 7 SCI groups were sacrificed postoperatively at 6 and 12 hours, and 1, 3, 5, 7, 14 days respectively. Western blot was used to detect changes of cyclin H protein expression after SCI.Immunohistochemical staining and immunofluorescence double staining were used to study the distribution and localization of cyclin H.Results Western blot analysis showed cyclin H protein level increased firstly and then decreased, reaching a peak at 3 days.Immunohistochemical analysis showed that cyclin H was distributed widely in a normal spinal cord, but at 3 days, its expression was up-regulated obviously in both white and grey matters.lmmunofluorescence double staining suggested that cyclin H was co-localized with NeuN, CD11b and GFAP.Conclusion The expression of cyclin H undergoes spatiotemporal changes after SCI, suggesting cyclin H may play a role in the SCI pathological process.