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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: To observe the changes and localization of insulin-like growth factor 2 (IGF2) and its receptor IGF2R protein in hippocampus at different time points after fimbria fornix cut. Methods: The quantities of IGF2 / Hoechst and IGF2R / Hoechst positive cells in the dentate gyrus and granulosa subtypes were detected by ELISA, Western blot and immunofluorescence histochemistry. Results: (1) The result of ELASA showed that the expression of IGF2 began to increase on the 7th day after cutting, reaching the highest level on the 14th day, then decreased slowly and dropped to the normal level on the 28th day. (2) Western blot results showed that IGF2R also had a corresponding increase and regression of the expression, indicating that there is a clear correlation between the two. (3) The results of immunofluorescence staining showed that the numbers of IGF2R positive cells in the dentate gyrus and granule lower layer of hippocampus in the normal group were less, and the number of IGF2R positive cells increased slightly on the 3rd day after fimbria fornix transection, but the difference was not obvious ; On the 7th day, the number of positive cells increased obviously; on the 14th day, the number of positive cells continued to increase and reached the highest level; the number of positive cells began to decrease on the 21st day and was close to the normal level on the 28th day. Conclusion: The high expression of IGF2R and its receptor IGF2R after fimbria fornix transection may be related to inhibiting the proliferation of neural stem cells and radial glial cells during hippocampal neurogenesis, thus initiating its differentiation into neurons.