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目的探讨自然衰老小鼠室管膜下区(SVZ)神经干细胞(NSCs)在衰老过程中生物学特点的改变。方法 2月龄、28月龄小鼠,各10只,体外分离、培养、鉴定NSCs,5-溴-2-脱氧尿苷(Brd U)染色比较两组细胞的增殖水平;β-半乳糖苷酶(SA-β-Gal)染色检测细胞衰老水平;细胞内活性氧(ROS)试剂盒检测细胞ROS表达水平;Western blotting检测细胞周期蛋白D1(cyclin D1)、p16及p19蛋白表达水平;体内用巢蛋白(Nestin)/性别决定基因高迁移率组蛋白-2(Sox2)检测两组小鼠SVZ区厚度的差异。结果年轻、年老小鼠的神经干细胞能表达Nestin及Sox2,符合神经干细胞的表达特性。与年轻小鼠相比,年老小鼠神经干细胞的Brd U阳性细胞比例显著下降,SA-β-Gal阳性细胞百分比显著升高,细胞活性氧水平显著升高,SVZ区厚度明显变薄,差异具有统计学意义(P<0.05)。在分子水平上表现为cyclin D1蛋白表达水平明显下降,p16、p19蛋白表达水平显著升高。结论年老小鼠NSCs出现增龄性变化,这些变化可能在大脑衰老中起重要作用。
Objective To investigate the changes of biological characteristics of neural stem cells (NSCs) in the subventricular zone (SVZ) of aging mice during senescence. Methods The 2-month-old and 28-month-old mice were randomly divided into 10 groups. The NSCs and BrdU staining were used to compare the proliferation of the two groups. Β-Galactoside The level of ROS was measured by enzyme-linked immunosorbent assay (ELISA). The expression of cyclin D1, p16 and p19 protein was detected by Western blotting. Nestin / sex-determining gene high-mobility group histone-2 (Sox2) was used to detect the difference of SVZ thickness between two groups of mice. Results The neural stem cells of young and old mice could express Nestin and Sox2, which accorded with the characteristics of neural stem cells. Compared with young mice, the percentage of Brd U positive cells in NSCs in aged mice decreased significantly, the percentage of SA-β-Gal positive cells increased significantly, the level of reactive oxygen species increased significantly, and the thickness of SVZ decreased significantly Statistically significant (P <0.05). At the molecular level, the expression of cyclin D1 protein decreased significantly, p16, p19 protein expression increased significantly. Conclusion Age-related changes of NSCs appear in aged mice, and these changes may play an important role in the brain aging.