切割穹窿海马伞大鼠海马IGF2及其受体IGF2R蛋白的表达变化  

The changes of IGF2/IGF2R protein expression in rat hippocampus after the fimbria /fornix transection

作　　者：王海琴[1] 金国华[1] 邹琳清[1] 秦建兵[1] 赵荷艳[1] 丁会[1] 陶学磊 

机构地区：[1]南通大学医学院人体解剖学系,江苏省神经再生重点实验室,南通226001

出　　处：《神经解剖学杂志》2013年第6期625-630,共6页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金(31271138);江苏省自然科学基金(BK2012659);江苏高校优势学科建设工程资助项目(PAPD)

摘　　要：目的:观察穹窿海马伞切割后不同时间点海马胰岛素样生长因子2(insulin-like growth factor,IGF2)及其受体IGF2R蛋白的表达变化及定位。方法:应用ELISA、Western blot和免疫荧光组织化学方法检测海马内IGF2及其受体IGF2R蛋白,以及齿状回门区和颗粒下层中IGF2/Hoechst和IGF2R/Hoechst阳性细胞的数量及形态学变化。结果:(1)ELASA结果显示:IGF2在切割后7 d表达开始上升,14 d达最高水平,随后缓慢下降,28 d时降至正常水平。(2)Western blot结果显示:IGF2R也存在一个相应的表达升高与消退的过程,说明两者具有明显的相关性。(3)免疫荧光染色结果显示:正常组海马齿状回门区和颗粒下层IGF2及其受体IGF2R阳性细胞的数量较少,切割穹窿海马伞后第3 d数量稍有增多,但差异不明显;7 d时阳性细胞明显增多;14 d时阳性细胞继续增多,并达到最高水平;21 d后阳性细胞的数量开始下降,28 d时接近正常组水平。结论:切割穹窿海马伞后IGF2及其受体IGF2R的高表达可能与海马神经再生中抑制神经干细胞和放射状胶质细胞增殖,从而启动其向神经元等分化有关。Objective： In order to observe the temporal and spatial changes of the insulin-like growth factor （IGF2） and its receptor IGF2R protein expression, and their location in rat hippocampus after fimbria/fomix transection at different time points. Methods： ELISA, Western blot and immunofluorescence histochemical methods were used to detect IGF2 and its receptor IGF2R protein expression in hippocampus, but also the number of IGF2/Hoechst and IGF2R/Hoechst positive cells and their morphologic change in hilus and subgranular layer of dentate gyms. Results： （ 1 ） The results of ELASA showed that the IGF2 expression began to up-regulate on the 7th day after transection, reaching a peak level on the 14th day, and then decreased slowly to the normal level on the 28th day. （2） Western blot results showed that IGF2R also had a corresponding process of rise and fade concerning its expression, suggesting that the correlation of IGF2 and IGF2R. （3） Immunofluorescence staining results showed that a few of IGF2 and IGF2R positive cells were expressed in hilus and subgranular layer of dentate gyms in control group. After the fimbria/fornix transection, the number of IGF2 andIGF2R positive cells showed a slight increase in number, but no significant difference was observed on the 3rd day; in- creased obviously on the 7th day; and continued to rise and peaked on the 14th day. Then the number of IGF/Hoechst- and IGF2R/Hoeehst-positive cells started to decrease 21st day later and closed to the normal level on the 28th day. Con- clusion ： That the up-regulated IGF2 and IGF2R in the fimbria/fomix transected hippoeampus may be related to inhibition of neural stem cells and the radical glial cells proliferation in the hippocampal neural regeneration, which launched its dif- ferentiation to neurons

关 键 词：胰岛素样生长因子2 胰岛素样生长因子2受体 海马 穹窿海马伞切割 免疫印迹 酶联免疫吸附 免 疫荧光 大鼠 

分 类 号：R338[医药卫生—人体生理学]