雌激素替代治疗对中老年卵巢切除大鼠海马CA1区有髓神经纤维的影响
发布时间：2018-07-17 19:17
【摘要】：雌激素是人体内主要的女性激素之一，，已有大量研究表明雌激素替代治疗对中枢神经系统认知功能具有保护作用。而雌激素替代治疗改善认知的同时对海马及其分区体积的影响目前多为影像学研究而尚未见组织学定量数据报告，且现有研究结果仍存在争议；此外现已知的雌激素替代治疗对认知功能的改善可以通过保护海马齿状回神经元存活和增殖，改善神经元兴奋性，增加海马CA1区树突棘和突触数量等方式来实现，而雌激素对中老年大鼠海马内传导神经冲动的有髓神经纤维的作用的精确定量研究目前尚未见相关报道。因此，本实验利用Morris水迷宫，冰冻切片技术，电镜技术及体视学方法研究雌激素替代治疗对中老年卵巢切除大鼠空间记忆能力，海马及其CA1体积，以及CA1区内有髓神经纤维的影响，并为雌激素替代治疗作用中老年卵巢切除大鼠行为学和海马形态学提供有力的数据基础和理论依据。 1.材料与方法 1.110月龄SD雌性大鼠切除双侧卵巢，术后一个月将大鼠随机分为雌激素替代治疗组（OVX＋E2组）（10只）和安慰剂组（OVX＋Veh组）（9只）。其中OVX＋E2组给予连续四周的17β-雌二醇皮下注射，OVX＋Veh组给予连续四周的安慰剂（芝麻油）皮下注射。雌激素替代治疗后，应用Morris水迷宫对其空间参考记忆能力进行测试。 1.2Morris水迷宫测试后，分别从OVX+Veh组和OVX+E2组中随机选取5只大鼠，灌注固定后随机选取一侧大脑半球作1mm连续切片，在解剖显微镜下放大10倍拍照；将切得的1mm脑片在冰冻切片机中速冻并切取一张10μm脑片做苏木素染色显示神经元，用以区分CA1区范围。然后利用体视学方法对海马及其CA1区体积进行测定。 1.3将已灌注固定的另一侧大脑半球作1mm连续切片，取出包含海马结构的脑组织切片，随机切取1mm3海马CA1区组织块，每只动物取5个标本块做成60nm的超薄切片，从每张切片上随机取15个视野在透射电镜下放大8000倍拍照。 1.4利用体视学方法分别对OVX+Veh组和OVX+E2组大鼠海马CA1区有髓神经纤维长度密度及总长度、有髓神经纤维体积密度及总体积、髓鞘体积密度及总体积、轴突体积密度及总体积以及平均直径进行定量研究。 2.结果 2.1OVX＋E2组大鼠Morris水迷宫的寻台潜伏期明显较OVX＋Veh组短（P＜0.05）。 2.2两组大鼠的海马总体积没有显著性改变（P=0.100）。 2.3两组大鼠的CA1区总体积没有显著性改变（P=0.366）。 2.4OVX＋E2组有髓神经纤维总体积较OVX＋Veh组显著增加了31.4%（P=0.016）。 2.5OVX＋E2组有髓神经纤维髓鞘总体积较OVX＋Veh组显著增加了37.8%（P=0.009）。 2.6OVX＋E2组有髓神经纤维轴突总体积较OVX＋Veh组增加了35.0%，两组间不存在显著性差异（P=0.087）。 2.7OVX＋E2组有髓神经纤维总长度较OVX＋Veh组增加了28.2%，两组间不存在显著性差异（P=0.105）。 2.8OVX＋E2组有髓神经纤维平均直径较OVX＋Veh组减少了1.3%，两组间不存在显著性差异（P=0.828）。 全文结论 1.本研究发现为期四周的雌激素替代治疗对中老年雌性卵巢切除SD大鼠的空间记忆能力具有显著的保护作用。 2.本研究发现为期四周的雌激素替代治疗能增加中老年雌性卵巢切除SD大鼠海马CA1区有髓神经纤维总体积及有髓神经纤维髓鞘总体积。
[Abstract]:Estrogen is one of the main female hormones in human body , and there is a great deal of research that estrogen replacement therapy has protective effect on the cognitive function of the central nervous system .
The effect of estrogen replacement therapy on the spatial memory capacity , hippocampus and CA1 volume of hippocampal CA1 region and myelinated nerve fibers in CA1 region were studied by Morris water maze , frozen section technique , electron microscope technique and stereology method .
1 . Materials and Methods
1.110 month - old SD female rats were randomly divided into estrogen replacement therapy group ( 10 rats ) and placebo group ( 9 rats ) .
After 1 . 2Morris water maze test , 5 rats were randomly selected from the group of the ovariectomized + Veh group and the ovariectomized + E2 group , and 1 mm continuous slices were randomly selected on one side of the cerebral hemisphere after the perfusion fixation , and the images were magnified 10 times under an anatomical microscope ;
A slice of 1 mm slices were quickly frozen in a frozen slicer and a 10.mu . m slices of brain slices were cut to show neurons to distinguish the CA1 region . Then , the volumes of the hippocampus and CA1 region were measured by stereology .
1.3 slices of the brain tissue containing the sea horse structure are taken out and the tissue blocks of the CA1 region of the hippocampus CA1 region of the hippocampus are taken out by taking out the slices of the brain tissue containing the sea horse structure , and each animal takes 5 specimen blocks to be made into ultrathin sections of 60nm , and 15 fields of view are randomly taken from each slice to be enlarged by 8000 times to take pictures under transmission electron microscope .
1.4 The density and total length of myelinated nerve fibers , the volume density and total volume of myelinated nerve fibers , the volume density and total volume of myelin sheath , the volume density and the total volume of myelinated nerve fibers and the mean diameter were studied by stereology .
2 . Results
2 . The latency period of the Morris water maze was shorter than that in the ovariectomized rats ( P < 0.05 ) .
2 . There was no significant change in the total volume of hippocampus in the two groups ( P = 0 . 100 ) .
2 . There was no significant change in the total volume of CA1 region in both groups ( P = 0.366 ) .
2 . The total volume of myelinated nerve fibers increased significantly ( P = 0 . 016 ) .
The total volume of myelinated myelinated myelinated myelinated myelinated myelinated myelinated myelinated myelinated myelinated myelinated myelinated myelinated myelinated myelinated myelinated myelinated myelinated myelinated myelinated myelinated myelinated nerve fibers increased by 38.8 % ( P = 0 . 009 ) .
There was no significant difference between the two groups ( P = 0.087 ) .
There was no significant difference between the two groups ( P = 0.105 ) .
There was no significant difference between the two groups ( P = 0.828 ) .
full - text conclusion
1 . The study found that the four - week estrogen replacement therapy had a significant protective effect on the spatial memory ability of female ovariectomized female SD rats .
2 . The study found that the four - week estrogen replacement therapy could increase the total volume of myelinated nerve fibers and the total volume of myelinated nerve fibers in CA1 region of SD rats .
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R749.16
[Abstract]:Estrogen is one of the main female hormones in human body , and there is a great deal of research that estrogen replacement therapy has protective effect on the cognitive function of the central nervous system .
The effect of estrogen replacement therapy on the spatial memory capacity , hippocampus and CA1 volume of hippocampal CA1 region and myelinated nerve fibers in CA1 region were studied by Morris water maze , frozen section technique , electron microscope technique and stereology method .
1 . Materials and Methods
1.110 month - old SD female rats were randomly divided into estrogen replacement therapy group ( 10 rats ) and placebo group ( 9 rats ) .
After 1 . 2Morris water maze test , 5 rats were randomly selected from the group of the ovariectomized + Veh group and the ovariectomized + E2 group , and 1 mm continuous slices were randomly selected on one side of the cerebral hemisphere after the perfusion fixation , and the images were magnified 10 times under an anatomical microscope ;
A slice of 1 mm slices were quickly frozen in a frozen slicer and a 10.mu . m slices of brain slices were cut to show neurons to distinguish the CA1 region . Then , the volumes of the hippocampus and CA1 region were measured by stereology .
1.3 slices of the brain tissue containing the sea horse structure are taken out and the tissue blocks of the CA1 region of the hippocampus CA1 region of the hippocampus are taken out by taking out the slices of the brain tissue containing the sea horse structure , and each animal takes 5 specimen blocks to be made into ultrathin sections of 60nm , and 15 fields of view are randomly taken from each slice to be enlarged by 8000 times to take pictures under transmission electron microscope .
1.4 The density and total length of myelinated nerve fibers , the volume density and total volume of myelinated nerve fibers , the volume density and total volume of myelin sheath , the volume density and the total volume of myelinated nerve fibers and the mean diameter were studied by stereology .
2 . Results
2 . The latency period of the Morris water maze was shorter than that in the ovariectomized rats ( P < 0.05 ) .
2 . There was no significant change in the total volume of hippocampus in the two groups ( P = 0 . 100 ) .
2 . There was no significant change in the total volume of CA1 region in both groups ( P = 0.366 ) .
2 . The total volume of myelinated nerve fibers increased significantly ( P = 0 . 016 ) .
The total volume of myelinated myelinated myelinated myelinated myelinated myelinated myelinated myelinated myelinated myelinated myelinated myelinated myelinated myelinated myelinated myelinated myelinated myelinated myelinated myelinated myelinated myelinated nerve fibers increased by 38.8 % ( P = 0 . 009 ) .
There was no significant difference between the two groups ( P = 0.087 ) .
There was no significant difference between the two groups ( P = 0.105 ) .
There was no significant difference between the two groups ( P = 0.828 ) .
full - text conclusion
1 . The study found that the four - week estrogen replacement therapy had a significant protective effect on the spatial memory ability of female ovariectomized female SD rats .
2 . The study found that the four - week estrogen replacement therapy could increase the total volume of myelinated nerve fibers and the total volume of myelinated nerve fibers in CA1 region of SD rats .
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R749.16
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