基于海马神经元线粒体自噬探讨脾气虚心神不足的可能机制  

Possible Mechanism Of Uneasiness Caused By Spleen Qi Deficiency Based On Hippocampal Neuron Mitophagy

作　　者：戴国英 李振钰 刘文俊[1] 许欣竹 王凌志[1] 单德红[1] DAI Guoying;LI Zhenyu;LIU Wenjun;XU Xinzhu;WANG Lingzhi;SHAN Dehong(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoing,China;Balinzuoqi Hospital of Mongolian Medicine and Chinese Medicine,Chifeng 025450,Inner Mongolia,China;Liaoning Vocational College of Medicine,Shenyang 110101,Liaoing,China)

机构地区：[1]辽宁中医药大学,辽宁沈阳110847 [2]巴林左旗蒙医中医院,内蒙古赤峰025450 [3]辽宁医药职业学院,辽宁沈阳110101

出　　处：《中华中医药学刊》2019年第11期2636-2638,I0013,共4页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81673851)

摘　　要：目的:通过检测脾气虚大鼠行为学及其海马神经元线粒体超微结构、基本功能和自噬水平,探讨脾气虚所致心神不宁的可能机制。方法:16只雄性SD大鼠随机分为正常组和脾气虚组(模型组),每组8只;旷场实验检测大鼠行为学;透射电镜观察海马神经元线粒体超微结构;JC-1法检测海马神经元线粒体膜电位;ELISA法检测海马神经元线粒体ATP和反应性氧簇(ROS)水平;Western blot法检测海马线粒体PTEN诱导激酶1 (PINK1)、Parkin、微管相关蛋白轻链3(LC3)-I和II表达。结果:与正常组相比,模型组大鼠在旷场的中心活动时间明显延长,周边活动时间缩短,运动距离和直立次数显著减少;海马神经元部分线粒体嵴消失,自噬样结构减少;海马神经元线粒体的ATP水平和膜电位下降明显,PINK1、Parkin和LC3-II表达及LC3-II/I比值均显著下降。结论:脾气虚所致心神不足与海马神经元线粒体损伤及其自噬水平低下有关。Objective: To study the possible mechanism of uneasiness caused by spleen Qi deficiency by testing behaviors and mitochondrial ultrastructure, basic function and mitophagy flux in hippocampal neurons. Methods: Sixteen male SD rats were randomly divided into the normal group and spleen Qi deficiency group(model group) with 8 rats in each group. The open-field test was used to observe behaviors and transmission electron microscope was chosen to observe hippocampal neuronal ultrastructures. JC-method was used to check the mitochondrial membrane potential. ELISA was applied to detect mitochondrial ATP and reactive oxygen species(ROS) levels. Western blot was employed to test expressions of PTEN-induced putative kinase 1(PINK1), Parkin, microtubule associated protein light chain(LC3)-I and II. Result: Compared with those in the normal group, rats in the model group spent more time in the center and less time in the periphery, covered shorter motion distances and reared less times. The hippocampal neuronal mitochondria exhibited decreased cristae and autophaged-like structures, lowered the ATP level and membrane potential, reduced expressions of PINK1, Parkin and LC3-II and the ratio of LC3-II/I. Conclusion: The uneasiness caused by spleen Qi deficiency might be caused by mitochondrial damage and mitophagy inhition in hippocampal neurons.

关 键 词：脾气虚 心主神明 海马神经元 线粒体自噬 行为学 

分 类 号：R2-03[医药卫生—中医学]