安寐丹对睡眠剥夺大鼠海马CA1区细胞结构及神经元损伤的保护作用  

Protective Effects of Anmeidan on Cell Structure Against Neuronal Damage in Hippocampal CA1 Region of Sleep-deprived Rats

作　　者：谢光璟 徐波[1] 夏婧 王平[1] XIE Guang-jing;XU Bo;XIA Jing;WANG Ping(Hubei University of Chinese Medicine,Wuhan 430065,China)

机构地区：[1]湖北中医药大学,武汉430065

出　　处：《中国实验方剂学杂志》2022年第7期26-32,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家重点研发计划“中医药现代化研究”重点专项(2018YFC1705600);国家自然科学基金青年基金项目(82104712)。

摘　　要：目的:探讨安寐丹对睡眠剥夺大鼠海马神经元及相关蛋白表达的影响。方法:通过随机数字表法将SD大鼠分为空白组、模型组、安寐丹组和褪黑素组。空白与模型组给予等容生理盐水,安寐丹组给药9.09 g·kg^(-1)·d^(-1)。褪黑素组给药0.27 g·kg^(-1)·d^(-1)。自制睡眠剥夺箱造模4周。Ethovision XT视频分析系统监测大鼠自发活动状态,苏木素-伊红(HE)染色观察海马CA1区神经元形态,尼氏染色观察海马CA1区神经元及尼氏体变化,超微电镜观察海马细胞超微结构,免疫组化检测海马CA1区胶质纤维酸性蛋白(GFAP)、微管相关蛋白2(MAP2)、巢蛋白(Nestin)、神经元特异核蛋白(NeuN)的表达。结果:与空白组比较,模型组中心点移动距离延长,平均活动速度增加,中心点累计持续时间增加,身体填充平均值增加,活动频率更高(P<0.01),海马CA1区神经元数量减少,排列紊乱,核皱缩,细胞质深染,尼氏体数量减少,出现明显丢失,线粒体出现肿胀变形,嵴缩短,高尔基体囊泡肿胀,GFAP蛋白积分吸光度(IA)增加,MAP2、Nestin、NeuN蛋白IA下降(P<0.01);与模型组比较,安寐丹组中心点移动距离缩短,平均活动速度降低,中心点累计持续时间缩短,身体填充平均值降低,活动频率降低(P<0.05,P<0.01),海马CA1神经元损伤减轻,细胞数量增加,核仁清晰,胞浆明显,尼氏体数量增加,线粒体等细胞器损伤减轻,GFAP蛋白IA降低,MAP2、Nestin、NeuN蛋白IA升高(P<0.05,P<0.01)。结论:安寐丹改善睡眠剥夺大鼠海马CA1神经元结构损伤,并可能通过降低GFAP蛋白表达,增加MAP2、Nestin、NeuN蛋白表达而实现。Objective:To investigate the effects of Anmeidan(AMD)on neuronal structure and neuronal marker protein expression in the hippocampal CA1 region of sleep-deprived(SD)rats.Method:Rats were randomly divided into control group,model group,an AMD group(9.09 g·kg^(-1)·d^(-1)),and melatonin group(0.27 g·kg^(-1)·d^(-1)).Rats in the control group and the model group received equal volumes of physiologicol saline.The SD model was induced by the self-made sleep deprivation box for four weeks.Ethovision XT system detected and analyzed the spontaneous behaviors of rats.The histomorphology of neurons in the hippocampal CA1 region was observed by hematoxylin-eosin(HE)staining and Nissl staining,and the changes in Nissl bodies were observed by Nissl staining.The ultrastructure of hippocampal cells was observed by transmission electron microscopy(TEM).Immunohistochemistry was used to detect the expression of glial fibrillary acidic protein(GFAP),microtubule-associated protein 2(MAP2),nestin,and neuronal nuclei(NeuN)in the CA1 region.Result:Compared with the control group,the model group showed longer distance,increased average activity speed,cumulative duration,average body fill,and higher activity frequency(P<0.01).Besides,the neurons in the CA1 region were reduced in number with disorganized arrangement,wrinkled nuclei,deeply stained cytoplasm,reduced Nissl bodies,swollen and deformed mitochondria,shortened cristae,and swollen Golgi vesicles.Furthermore,the mean integral absorbance(IA)value of GFAP increased and those of MAP2,nestin,and NeuN decreased(P<0.01).Compared with the model group,the AMD group showed shortened distance traveled,lower average activity speed,shorter cumulative duration,decreased average body fill,and reduced activity frequency(P<0.05,P<0.01).Moreover,the neurons in the CA1 region were relieved from damage with increased cell number,clear nuclei and cytoplasm,increased Nissl bodies,and relieved mitochondrial damage.The IA value of GFAP decreased and those of MAP2,nestin,and NeuN increased(P<0.05,P<0.0

关 键 词：安寐丹 睡眠剥夺 胶质纤维酸性蛋白(GFAP) 微管相关蛋白2(MAP2) 巢蛋白(Nestin) 神经元特异核蛋白(NeuN) 

分 类 号：R2-0[医药卫生—中医学] R22