【作者】 郑雪；

【导师】 赵善廷；

【作者基本信息】 兰州大学 ， 动物学， 2013， 硕士

【摘要】 目前成体神经干细胞在临床应用中具有广阔的前景,对它的研究一直是国内外神经领域研究的热点,大量的形态学和行为学实验表明成体干细胞与学习记忆密切相关。近年来砷中毒对中枢神经系统的损害已引起人们的高度重视,而有关慢性砷中毒成年小鼠动物模型成体神经发生的研究尚未见报道。本实验通过建立慢性砷中毒小鼠模型,着重应用免疫组化技术在细胞水平上研究三氧化二砷中毒对海马结构中的齿状回颗粒下层(sub granular zone, SGZ区)的成体神经发生的影响,包括成体干细胞的增殖速度和新生细胞的存活,为阐明成体神经发生与慢性砷中毒及学习和记忆的关系提供必要的理论和实验基础。目的：探讨三氧化二砷对小鼠海马成体神经发生的影响。方法：用C57小鼠建立砷中毒模型,和BrdU(5-溴脱氧尿嘧啶核苷)腹腔注射、活体灌注取脑、常规冰冻切片等方法,并用BrdU免疫组化染色结合普通光学显微镜进行显微观察和照相,计数小鼠海马单位长度齿状回内颗粒细胞层中BrdU阳性细胞数并进行统计学分析。结果：BrdU注射后存活24小时处死的空白对照组(NS) BrdU阳性细胞平均数是17.63个,低剂量组(1mg/kg)BrdU阳性细胞平均数是21.17个,高剂量组(2mg/kg) BrdU阳性细胞平均数是11.50个；BrdU注射后存活28天处死的空白对照组(NS) BrdU阳性细胞平均数是11.40个,低剂量组(1mg/kg) BrdU阳性细胞平均数是5.40个,高剂量组(2mg/kg) BrdU阳性细胞平均数是5.57个。结论：三氧化二砷在一定的浓度范围内,可引起海马齿状回成体神经干细胞数量的增加,促进其成体神经发生,但其会抑制新生细胞的存活。更多还原

【Abstract】 Adult neural stem cells in clinical applications has broad prospects, its research has been a hotspot of neurological research in the world, a large number of morphological and behavioral experiments show that adult stem cells and learning and memory are closely related. In recent years, the arsenic poisoning damage to the central nervous system has aroused great attention, but the research of chronic arsenic poisoning of adult neurogenesis in adult mouse model has not been reported. Our experiment established chronic arsenic poisoning mice model and focuses on the cellular level study of the hippocampal formation in the subzone of dentate gyrus (SGZ), including the proliferation of adult stem cells speed and newborn cell viability, to clarify the relationship of the adult neurogenesis and chronic arsenic poisoning and the learning and memory,to provide the necessary theoretical and experimental basis. Objective:To explore the effect of arsenic trioxide on the adult neurogenesis of mice. Methods:The C57mice were treated with two different doses of arsenic trioxide by intraperitoneal injection.4weeks after administration, the mice received intraperitoneal injection of5-Bromo-2’-deoxyuridine and then were perfused with4%paraformeldhyde. The fixed brains were sliced by using cryotome. The sections were stained for mmunocytochemistry against BrdU to investigate quantificationally the proliferation of adult neural stem cells and survival of newly generated cells in the SGZ of the dentate gyrus. The density of BrdU-positive cells in the dentate gyrus were counted unter microscope and evaluated by SPSS17.0software. Results:In the group perfused24hours after BrdU injection, the mean number of control group(NS) is17.63, the low dose group(1mg/kg) is21.17, the high dose group(2mg/kg) is11.50; In the group perfused28days after BrdU injection, the mean number of control group(NS) is11.40, the low dose group(lmg/kg) is5.40, the high dose group(2mg/kg) is5.57.Conclusion:In a certain dose range, As2O3exposure induced cell proliferation, but impaired the survival of newly generated cells in the SGZ of the dentate gyrus in mice.更多还原