慢性束缚应激小鼠海马应激性病理效应.docxVIP

慢性束缚应激小鼠海马应激性病理 效应 【摘要】目的观察慢性束缚应激下小 鼠海马神经元数目的改变及天冬氨酸特异 性半光氨酸蛋白酶3的活性，探讨应激对海 马的损害作用及途径。 方法 将16只小鼠随 机分为对照组和应激组各 8只，采用50ml 离心管建立慢性束缚应激模型，每晚束缚 8min o 14d后处死，取小鼠海马常规固定制 HE片、免疫组化天冬氨酸特异性半光氨酸蛋 白酶3片,在显微镜下进行细胞记数并显微 拍照。结果 慢性束缚应激组小鼠海马数目 较对照组减少，同时天冬氨酸特异性半光氨 酸蛋白酶3阳性细胞率增加（）；镜下神经元 发生空泡变性，存在典型凋亡细胞。结论慢 性束缚应激海马神经元凋亡蛋白天冬氨酸 特异性半光氨酸蛋白酶 3活性增加，天冬氨 酸特异性半光氨酸蛋白酶 3可能参与了应激 状态下海马的细胞凋亡。 【关键词】 束缚应激；海马；天冬氨 酸特异性半胱氨酸蛋白酶 3;病理形态学 Stressful pathological effects of hippocamal in chronic restraint stress mice [Abstract 】 Objective To observe changes of hippocamal neurons and activities of Caspase3 in the hippocampus of chronic restraint stress mice and explore the injuring actions and ways of stress to hippocampus. Methods 16 mice were randomly divided into control and stressful group(both n=8),RST devices were constructed with 50ml polypropylene conical tubes, restrain stress was imposed for 8 minutes per night. After Fourteen days mice were killed and brains were taken. The paraffin section for the He and immunohistochemistry Caspase3 were routinely made, counts of positive cell and photomicrographs taken under microscope. Results Decrease of hippocamal neurons were more marked in the stress than in the control group() and rate of positive cell of Caspase3 increased more significantly(); Neurons developed vacuolar degeneration and had dead cells. Conclusion Activity of hippocamal Caspase3 increased and Caspase3 might play a role in the death of hippocamal neurons under stress. 【Keywords 】 Restraint stress; hippocampus ;Caspase3; pathomorphism. 慢性持久的不良刺激对大脑可产生病 理生理损害。流行病学资料显示，慢性应激 与很多精神障碍如复发性抑郁症和创伤后 应激障碍(posttraumatic stress disorder,PTSD)的发病密切相关:1,2 ］。国 外有资料表明，慢性应激可以导致动物海马 CA1和CA3区锥体细胞形态结构受损，并认为 这可能与随后发生的精神障碍有关。 新近的 研究发现:3:,天冬氨酸特异性半光氨酸 蛋白酶3(Caspase3)参与了神经退行性疾病 的发生。本研究通过建立慢性束缚应激模 型，观察海马脑组织的形态学改变，并半定 量检测海马Caspase3活性，探讨慢性应激 对海马的病理损害。 1材料与方法1试剂器材兔抗小鼠 Caspase3抗，购于武汉博士德公司； ULTRASENSITIVETMS?敏试剂盒，购于福建 迈新生物技术开发公司；75%酉精，4嗾聚 甲醛，DAB显色液，50ml离心管，奥林巴斯 显微摄像显微镜，COINC光学显微镜等。2分 组处理昆明小白鼠，周龄4?6w,体重 19土 2g,由苏州大学试验动物中心提供。将 实