中华神经外科杂志
中華神經外科雜誌
중화신경외과잡지
Chinese Journal of Neurosurgery
2014年
2期
194-199
,共6页
张双伟%黄克伦%陈毕%滕红林%王靖%朱旻宇
張雙偉%黃剋倫%陳畢%滕紅林%王靖%硃旻宇
장쌍위%황극륜%진필%등홍림%왕정%주민우
脊髓损伤%星形胶质细胞%增殖
脊髓損傷%星形膠質細胞%增殖
척수손상%성형효질세포%증식
Spinal cord injury%Astrocytes%Proliferation
目的 观察脊髓损伤后p27kip1和Skp2蛋白表达及其意义.方法 改良Allen's法制作脊髓损伤模型,通过Western blot测定损伤后各时间段p27kip1和Skp2蛋白水平表达变化,免疫组织化学、免疫荧光双标,免疫共沉淀检测p27kip1和Skp2细胞定位.结果 Western blot结果显示脊髓损伤后,p27kip1显著地下调,Skp2上调,时间上二者的表达存在负相关.免疫组织化学和免疫荧光显示,空间上p27kip1和Skp2主要定位在广泛增殖的星形胶质细胞上.免疫共沉淀结果显示p27kip1和Skp2在脊髓中存在直接相互作用,且在损伤3d后作用加强.结论 脊髓损伤后Skp2参与介导的p27kip1表达下调,这与调节脊髓损伤后星形胶质细胞的增殖相关.
目的 觀察脊髓損傷後p27kip1和Skp2蛋白錶達及其意義.方法 改良Allen's法製作脊髓損傷模型,通過Western blot測定損傷後各時間段p27kip1和Skp2蛋白水平錶達變化,免疫組織化學、免疫熒光雙標,免疫共沉澱檢測p27kip1和Skp2細胞定位.結果 Western blot結果顯示脊髓損傷後,p27kip1顯著地下調,Skp2上調,時間上二者的錶達存在負相關.免疫組織化學和免疫熒光顯示,空間上p27kip1和Skp2主要定位在廣汎增殖的星形膠質細胞上.免疫共沉澱結果顯示p27kip1和Skp2在脊髓中存在直接相互作用,且在損傷3d後作用加彊.結論 脊髓損傷後Skp2參與介導的p27kip1錶達下調,這與調節脊髓損傷後星形膠質細胞的增殖相關.
목적 관찰척수손상후p27kip1화Skp2단백표체급기의의.방법 개량Allen's법제작척수손상모형,통과Western blot측정손상후각시간단p27kip1화Skp2단백수평표체변화,면역조직화학、면역형광쌍표,면역공침정검측p27kip1화Skp2세포정위.결과 Western blot결과현시척수손상후,p27kip1현저지하조,Skp2상조,시간상이자적표체존재부상관.면역조직화학화면역형광현시,공간상p27kip1화Skp2주요정위재엄범증식적성형효질세포상.면역공침정결과현시p27kip1화Skp2재척수중존재직접상호작용,차재손상3d후작용가강.결론 척수손상후Skp2삼여개도적p27kip1표체하조,저여조절척수손상후성형효질세포적증식상관.
Objective By studying the expression and distribution of p27kip and Skp2 in the rat models of contused spinal cord injury (SCI),to explore their biological fhnctions in the process of spinal cord injury and repair.Methods In this study,we performed the SCI model in adult Sprague Dawley rats with the improved Allen's method,investigated the expression profiles of p27kip and Skp2 following the SCI by western blot analysis.To further characterize the specific cell types expressing p27kip and Skp2,immunohistochemisty and immunofluorescent staining were performed with the cell-specific markers:NeuN (neuronal marker) and GFAP (astrocytic marker).Results The western blot analysis showed the spinal cord injury may induce p27kip1 and Skp2 dynamic changes,a significant down-regulation of p27kip1 and a concomitant up-regulation of Skp2 following spinal injury.There was a negative correlation in the expression of p27kip1 and Skp2 in a time-dependent manner.Double immunofluorescence staining showed that p27kip1 and Skp2 were co-expressed with NeuN and GFAP,and their expression were increased predominantly in astrocytes after injury,which were regenerating axons and largely proliferated after injury.Furthermore,coimmunoprecipitation studies demonstrated increased interactions among p27kip1 and Skp2 in the spinal cord after iujury.Conclusions The up-regulated expression of Skp2 after spinal cord injury,wbich related to the down-regulation of p27kip1,this event may involved in the proliferation of astrocytes after SCI.These results provide new insights into the molecular mechanisms underlying astrocyte proliferation during SCI and suggest that Skp2 and p27kip1 might play crucial roles in pathophysiology after SCI.
