摘要
目的:分析穹隆-海马伞切断加卵巢切除大鼠大脑皮质区、海马CA1区雌激素受体α与淀粉样β蛋白前体β位点裂解酶共存表达的变化。方法:实验于2003-03/12在青岛大学医学院附属医院脑病防治重点实验室完成。成年健康雌性Wistar大鼠14只,随机分为穹隆-海马伞切断加卵巢切除组和对照组,每组7只。穹隆-海马伞切断参照Klein等的方法,动物麻醉固定于脑立体定位仪后,参照Paxinos等大鼠脑立体定位图谱于前囟后2.2～2.5mm,中线外1.0mm处凿开颅骨,切开硬脑膜,自制双刃刀完全切断海马伞外侧缘。双侧卵巢切除参照Singh等的方法,麻醉动物后,打开腹腔,在肾脏下方的脂肪内取出卵巢,结扎并切除。对照组除不切断穹隆-海马伞和不切除卵巢外,其余步骤同穹隆-海马伞切断加卵巢切除组。免疫荧光双标细胞化学法染色,激光共聚焦显微镜下观察大鼠大脑皮质区、海马CA1区雌激素受体α、淀粉样β蛋白前体β位点裂解酶阳性细胞表达以及共存表达的变化。结果:14只大鼠参加实验,中途无脱落,均进入结果分析。①穹隆-海马伞切断加卵巢切除组大鼠皮质区绿色雌激素受体α阳性细胞数与对照组比较明显减少,差异有显著性意义[(4.42±3.99),(9.71±4.53),P<0.05];红色淀粉样β蛋白前体β位点裂解酶阳性细胞数量显著增多,差异有显著性意义[(20.00±5.16),(5.71±3.14),P<0.05];共表达双标阳性细胞数量显著增多,差异有显著性意义[(4.00±2.94),(1.14±1.67),P<0.05]。②穹隆-海马伞切断加卵巢切除组大鼠在海马CA1区雌激素受体α阳性细胞数与对照组比较明显减少,差异有显著性[(5.14±3.80),(12.57±3.59),P<0.01];淀粉样β蛋白前体β位点裂解酶阳性细胞数量明显增多,差异有显著性意义[(17.14±4.45),(6.85±1.95),P<0.01];共表达阳性细胞数无显著变化意义[(2.01±2.08),(2.00±2.08),P>0.05]。结论:穹隆-海马伞切断与卵巢切除可以导致大鼠大脑皮质区、海马CA1区雌激素受体α表达减少、淀粉样β蛋白前体β位点裂解酶表达增多,皮质区共存细胞表达增多,说明雌激素与淀粉样β蛋白前体代谢系统存在内在的联系。
AIM： To analyze the coexist expressive variety of estrogen receptor alpha （ERα） and beta-site beta-amyloid precursor protein （APP） cleaving enzyme （BACE） in the cerebral cortex and hippocampal CA1 region of the rats with fimbria/fornix transection and ovariectomy.
METHODS： The experiment was carried out in the Institute of Cerebrovascular Disease, Medical College Hospital of Qingdao University from March to December 2003. Fourteen healthy adult female Wistar rats were randomly divided into fimbria/fornix transection and ovariectomized group （n=7） and control group （n=7）. The rats were anaesthetized and the bilateral fimbria-fornix of brain were transected in the stereotaxic apparatus according to Klein＇s methods. After opening the skull at the coordinates at the bregma anterior-posterior 2.2-2.5 mm and lateral 1.0 mm according to the Paxinos＇ methods, the dorsal fornix, the anterior part of hippocampus and the fimbria were cut off with a double blade under visual inspection. About the ovariectomy, the rats were under anaesthesia, after the abdomen opened, bilateral ovaries in the fattiness under the kidney were ablated in order to erect the model according to Singh＇s methods. Control animals were sham-operated by opening the skull only and cut the abdomen only. The cells of ERa and BACE were stained by immunofluorescent double labeling and observed with confocal laser scanning microscopy. The number of masculine cells was counted under microscope at random selected 5 fields in every detected area, and then compared the average number of ERa and BACE of the control group.
RESULTS： All the 14 rats were involved in the analysis of results without deletion. （1） In the cerebral cortex of rats in the fimbria/fornix transection and ovariectomized group, the amount of the immunofluorescent green cells of ERα was remarkably decreased as compared with that in the control group （4.42±3.99, 9.71±4.53,P 〈 0.05）, and the amount of the immunofluorescent red cells of BACE was remarkably improved as compared with that in the control group （20.00±5.16, 5.71±3.14, P 〈 0.05）. The immunofluorescent double labeling positive cells in the cerebral cortex of fimbria/fornix transection and ovariectomized group was significantly increased as compared with that in the control group （4.00±2.94, 1.14±1.67, P 〈 0.05）. （2） In the hippocampal CA1 region of rats in the fimbria/fornix transection and ovariectomized group, the amount of the immunofluorascent green cells of ERα was remarkably decreased as compared with that in the control group （5.14±3.80, 12.57±3.59, P 〈 0.01）, and the amount of the immunofluorescent red cells of BACE was remarkably improved as compared with that in the control group （17.14±4.45, 6.85±1.95, P 〈 0.01）. There was no difference in the immunofluorescent double labeling cells in the hippocampal CA1 between the two groups （2.01±2.08, 2.00±2.08, P 〉 0.05）.
CONCLUSION： Fimbria/fornix transection and ovafiectomy can cause the expressions of BACE increased and the ERα decreased in hippocampal CA1 and cerebral cortex of rats, as well as the coexist cells increased, indicating that there is a relationship between the estrogen system and the system of beta-amyloid precursor protein.
出处
《中国临床康复》
CSCD
北大核心
2006年第14期74-76,共3页
Chinese Journal of Clinical Rehabilitation
基金
山东省科技厅科研基金资助项目(003130110
22130109)~~
