槲皮素抑制中枢神经突触前兴奋性依赖的胞吞动力学  

The Quercetin-modulated Activity-dependent Inhibition of Endocytosis at a Central Synapse

作　　者：李顺 高一鸣[2,3] 徐玥 胡佳琦 汤雯絮 孙小荃 薛磊 王万春 LI Shun;GAO Yi-Ming;XU Yue;HU Jia-Qi;TANG Wen-Xu;SUN Xiao-Quan;XUE Lei;WANG Wan-Chun(Jiangxi University of Traditional Chinese Medicine,Nanchang 330004,China;State Key Laboratory of Medical Neurobiology and MOE Frontiers Center for Brain Science,Fudan University,Shanghai 200433,China;Department of Physiology and Neurobiology,School of Life Sciences,Fudan University,Shanghai 200438,China;Research Institute of Intelligent Complex Systems,Fudan University,Shanghai 200433,China;Center for Rehabilitation Medicine,Department of Pain Management,Zhejiang Provincial People’s Hospital,Affiliated People’s Hospital,Hangzhou Medical College,Hangzhou 310014,China;Department of Traditional Chinese Medicine Surgery,Affiliated Hospital of Jiangxi University of Chinese Medicine,Nanchang 330006,China)

机构地区：[1]江西中医药大学,南昌330004 [2]复旦大学医学神经生物学国家重点实验室及脑科学教育部前沿研究中心,上海200433 [3]复旦大学生命科学学院生理与神经生物系,上海200438 [4]复旦大学智能复杂系统研究所,上海200433 [5]浙江省人民医院康复医学中心,疼痛科,杭州医学院附属人民医院,杭州310014 [6]江西中医药大学附属医院中医外科,南昌330006

出　　处：《生物化学与生物物理进展》2023年第6期1391-1402,共12页Progress In Biochemistry and Biophysics

基　　金：国家重点研发计划(2022YFC3602700,2022YFC3602702);科技创新2030-脑科学与脑启发智能项目(2021ZD0201301);国家自然科学基金(81960874,31971159);上海市教委创新计划(2019-01-07-00-07-E00041);江西省自然科学基金(20202ACB206010);上海市重大科技专项(2018SHZDZX01);张江实验室和上海脑科学与类脑研究中心资助项目。

摘　　要：目的槲皮素是一种广泛分布于药用植物中的黄酮类化合物,传统被认为具有神经保护作用。本研究利用位于大鼠脑干花萼状突触的突触前神经末梢进行膜片钳记录,研究槲皮素调控突触传递和可塑性的突触前机制。方法利用全细胞膜片钳结合膜电容记录,在突触后记录微小兴奋性突触后电流(m EPSC),在突触前神经末梢记录钙内流和神经囊泡的释放、回收以及可立即释放库(RRP)的恢复动力学。并且利用纤维刺激在轴突给予5~200 Hz的刺激,诱发突触后EPSC,记录突触后短时程抑制(STD)。结果100μmol/L槲皮素不影响突触后m EPSC的振幅、频率以及AMPA受体的动力学特征。在突触前神经末梢,槲皮素不改变钙内流或囊泡的释放,但显著抑制胞吐后网格蛋白依赖的慢速胞吞。抑制胞吞会导致突触前囊泡动员的减慢,降低RRP的补充速率,并且增强高频刺激下的短时程可塑性STD。结论本研究为槲皮素调控中枢神经突触传递提供全新的突触前神经机制,槲皮素有助于抑制中枢神经过度兴奋,进而发挥神经保护作用。Objective Quercetin,a flavonol compound widely distributed in fruits,vegetables,and medicinal plants,has been suggested to act as a neuroprotective agent.In the present study,we investigated the presynaptic effect of quercetin on synaptic transmission and plasticity.Methods Using whole-cell patch clamp and capacitance measurement technique,we recorded miniature excitatory postsynaptic currents(mEPSC),presynaptic calcium influx,vesicle release and recycling,and the replenishment of readily releasable pool(RRP).Additionally,we stimulated the axon with 5-200 Hz and recorded short-term depression(STD)in the postsynaptic neuron.Results We found that 100μmol/L quercetin in the extracellular solution did not affect the mEPSC amplitude and frequency,indicating a presynaptic mechanism modulating synaptic transmission.At the presynaptic nerve terminals,100μmol/L quercetin did not induce notable changes in calcium influx or vesicle release,but significantly inhibited clathrindependent slow endocytosis following exocytosis.The inhibition of endocytosis led to a slowdown of vesicle mobilization during stimulation,a reduction in readily releasable pool replenishment after stimulation,and enhancement of short-term depression during high-frequency repetitive stimulation in the postsynaptic principal neurons.Conclusion These results provide new insights into quercetin-modulated neuronal signaling and suggest a protective effect that prevents excessive excitatory synaptic transmission in brain circuits.

关 键 词：槲皮素 突触传递 囊泡胞吐 囊泡胞吞 电容测量 

分 类 号：Q4[生物学—生理学] Q6