摘要
目的 观察第10号染色体缺失与张力蛋白同源的磷酸酯酶基因（PTEN）/哺乳动物雷帕霉素靶蛋白（mTOR）信号通路在大鼠脊髓发育过程中的表达模式.方法 选取不同发育阶段（孕15 d子鼠记为A组,出生当天大鼠记为B组,出生后7d大鼠记为C组,出生后56 d大鼠记为D组）的SD大鼠,取出其脊髓,采用Western blot法检测该通路上关键蛋白的表达水平;同时,采用免疫荧光法检测各相应蛋白在大鼠脊髓的细胞定位.结果 Western blot结果显示,A、B、C、D组PTEN与内参肌动蛋白（actin）的灰度值比分别为0.281±0.086、0.462 ±0.002、0.591±0.022、0.392±0.001,A组与其他3组比较差异有统计学意义（P＜0.05）,D组与B、C组比较差异有统计学意义（P＜0.05）;A、B、C、D组S6与actin的灰度值比分别为0.646 ±0.116、0.450±0.129、0.824±0.082、0.174±0.038,D组与A、C组比较差异有统计学意义（P＜0.05）.免疫荧光染色结果显示脊髓神经元有丰富的PTEN与S6表达.结论 这些结果提示PTEN/mTOR信号通路可能与发育相关的表达模式,从而为中枢神经系统再生能力随着发育过程而降低这一现象提供依据.
Objective To observe the expression pattern of phosphatase and tensin homologue deleted on chromosometen （PTEN）/mammalian target of rapamycin （mTOR） pathway in the spinal cord of SD rats.Methods We dissected spinal cords of rats at developmental stages E15,P0,P7 and P56,and applied Western blotting to detect the expression levels of critical target proteins on the pathway.Furthermore,immunofluorescence was used to detect the cellular localization of these targets in the spinal cord.Results The ratio of gray value for PTEN to actin was 0.281 ± 0.086,0.462 ± 0.002,0.591 ± 0.022 and 0.392 ± 0.001 from E15 to P56.There was statistically significant difference between each two groups except for P0 and P7.The ratio of gray value for S6 to actin was 0.646 ± 0.116,0.450 ± 0.129,0.824 ± 0.082 and 0.174 ± 0.038 from E15 to P56.There was statistically significant difference between P56 to E15 and P7.Meanwhile neurons in the spinal cord of rats were found to be abundant of PTEN and S6 expression with immunofluorescence method.Conclusion Our data showed a development-related expression pattern of PTEN/mTOR signaling pathway in the spinal cord of rats.These results may provide a factual basis for the etiology of the decrease of intrinsic regeneration ability of central nervous system （CNS） along with development,and supply some thread for therapy after spinal cord injury.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2015年第3期588-590,共3页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金资助项目（30973028、30973044）
