新生小鼠胰岛增殖及口袋蛋白家族、E2F转录因子表达变化  

Changes of the proliferation of pancreatic islet, pocket protein family and E2F transcription factors during neonatal period in mice

作　　者：朱嘉瑜[1] 王晓兢[2] 张勤凤[2] 徐一力[2] 吕颖钺[2] 张洁[2] 吴雨洁[3] 武晓泓[2] 

机构地区：[1]江苏省江阴市人民医院内分泌科 [2]南京医科大学第一附属医院内分泌科,210029 [3]南京医科大学第一附属医院血液科,210029

出　　处：《中华糖尿病杂志》2013年第11期678-682,共5页CHINESE JOURNAL OF DIABETES MELLITUS

基　　金：国家自然科学基金项目（30971404、81070621、81261120566）;江苏省国际科技合作计划（BZ2011042）;江苏省医学重点人才项目（RC2011068）;江苏省“六大人才高峰”项目（苏人社发[2011]582号）

摘　　要：目的 观察新生小鼠胰岛细胞、细胞周期和视网膜母细胞瘤蛋白（Rb）、视网膜母细胞瘤样蛋白1（p107）、视网膜母细胞瘤样蛋白2（p130）及E2F转录因子的变化特点,探讨其与胰岛细胞增殖的关系.方法 采用免疫组化法检测出生第1、3和8周小鼠胰岛β/α细胞量、单个细胞面积和细胞数量的变化.采用胶原酶消化法分离纯化小鼠胰岛,以流式细胞术检测细胞周期,实时定量PCR及蛋白印迹技术检测Rb、p107、p130和E2F转录因子mRNA和蛋白表达.结果 （1）小鼠3周和8周胰岛β/α细胞量显著高于1周（P＜0.05）;单个β/α细胞面积在3周时无明显差异,8周时扩大1.7倍（P＜0.01）;而β/α细胞数量在3周增加约3.5/3.7倍（P＜0.01）,8周与3周无明显差异.（2）1周和3周时G0/G1期细胞比8周明显减少（81.3±1.2、82.8±3.3比92.6±1.3,F=6.5,P＜0.05）,而G2/M期显著增多（13.4±1.4、12.2±1.8比5.5±0.5,F=6.3,P＜0.05）,细胞增殖指数明显增加（P＜0.05）.（3）8周时Rb mRNA表达明显低于1周和3周（P＜0.01）;3周和8周时p107 mRNA显著低于1周（P＜0.05）,而p130 mRNA显著高于1周（P＜0.05）,E2F-1、E2F-2 mRNA均显著低于1周（均P＜0.01）;8周时E2F-3 mRNA显著低于1周（P＜0.05）;而E2F-4、E2F-5 mRNA在3周和8周时表达增加.（4）8周时Rb和p107蛋白表达均比1周明显降低,p130表达显著增高,3周和8周时E2F1表达显著降低（均P＜0.05）.结论 新生小鼠胰岛细胞量扩增明显,以细胞数量增加为主,细胞增殖显著.Rb、p107、p130及其相关E2F转录因子的表达存在动态变化,可能参与细胞增殖的调控.Objective To observe the changes of β/α cells,cell cycle,pocket protein family （retinoblastoma protein （Rb）,retinoblastoma-like protein 1 （p107）,retinoblastoma-like protein 2 （p130）） and E2F transcription factors of pancreatic islets in mice during neonatal period and to explore their relationship with the proliferation of islet cells.Methods β/α cell mass,cell size and cell number of mice islets at 1-,3-to 8-week after birth were detected by immunohistochemistry.Pancreatic islets were digested and isolated by collagenase.Cell cycle was detected by flow cytometry.The expression of mRNA and protein of Rb,p107,p130 and E2F transcription factors was detected by real-time quantitative polymerase chain reaction（PCR） and Western blotting.Results （1） Compared with 1 w,β/α cell mass significantly increased at 3 w and 8 w （P ＜ 0.05） ; cell size didn＇t change obviously at 3 w,but increased 1.7 times at 8 w （P ＜ 0.01） ; cell number increased nearly 3.5/3.7 times at 3 w （P ＜ 0.01）,and there＇s no significant difference between 8 w and 3 w.（2）Compared with 8 w,there were less cells in the G0/G1 phase and more cells in the G2/M phase at 1 wand3 w （81.3±1.2,82.8±3.3 vs92.6±1.3,F=6.48,P ＜0.05 ; 13.4 ± 1.4,12.2 ± 1.8 vs 5.5 ±0.5,F =6.25,P ＜0.05）,and the proliferation index were significantly higher at 1 w and 3 w （P ＜ 0.05）.（3） The mRNA level of Rb at 8 w was much lower than that at 1 w and 3 w （P ＜0.01） ; At 3 w and 8 w,the mRNA level of p107 were lower than that at 1 w （P ＜0.05）,whereas the level of p130 were much higher than that at 1 w （P ＜ 0.05）,and the level of E2F-1 and E2F-2 were significantly lower than that at 1 w （P ＜ 0.05）,but the mRNA expression of E2F-4 and E2F-5 were increased.Compared with 1 w,the protein level of Rb and p107 were much lower,but the level of p130 was significantly increased at 8 w （P ＜ 0.05）.Conclusion The β/α cell mass increases significantly during neonatal period in mice,primarily in c

关 键 词：胰岛 增殖 新生期 口袋蛋白家族 E2F转录因子 

分 类 号：R587.1[医药卫生—内分泌]