β淀粉样蛋白寡聚体毒性环境下Bryostatin-1对神经突触活性的影响和潜在机制  

Bryostatin-1 protects synapse morphology and function against Aβtoxicity

作　　者：潘文阳 陈鹏 孙邈 包小峰 PAN Wenyang;CHEN Peng;SUN Miao;BAO Xiaofeng(Department of Pharmacology,Nantong University,Nantong 226019,China)

机构地区：[1]南通大学药学院,南通226019

出　　处：《生物学杂志》2024年第3期85-91,共7页Journal of Biology

基　　金：国家自然科学基金项目(No.81801080,No.82071174);南通市科学技术基金项目(No.JC22022043);南通市第六期江海英才市级培养专项。

摘　　要：在原代海马神经元上建立通过外加Aβ寡聚体所导致的突触毒性模型,并使用蛋白印迹、激酶活性、激光共聚焦显微成像和电生理等技术检测PKC激酶的非选择性激动剂Bryostatin-1是否可以保护Aβ寡聚体导致的突触毒性。结果显示,在Aβ毒性模型中,Bryostatin-1可以改善树突棘的形态,并促进树突棘成熟。此外,电生理结果显示Bryostatin-1可以显著改善突触后的微型兴奋性突触后电流频率,AMPA受体与NMDA受体的比例并没有发生显著改变。蛋白印迹结果显示Bryostatin-1可以上调突触生物标记物水平。蛋白印迹试验结果表明,Bryostatin-1可以上调mTOR-S6K1信号通路的活性。通过在神经元中转染GFP-RFP-LC3质粒并进行激光共聚焦显微观察,结果提示Aβ寡聚体可以抑制神经元内自噬通量,而Bryostatin-1可以提高被Aβ寡聚体所抑制的自噬通量。结果提示,Bryostatin-1激活mTOR-S6K1信号通路,并随后调节细胞自噬功能可能是其在Aβ寡聚体毒性环境下保护神经元突触功能的分子机制。By using Western Blot,kinase activity assay,confocolmicroscopy,and electrophysiology recording methods,the synapse toxicity induced by Bryostatin-1 protects Aβoligomers was studied and its potential mechanisms was investigated.Results showed that in the present of Aβoligomers,Bryostatin-1 could significantly protect the morphology of dendritic spines,and promote the maturation of dendritic spines.Furthermore,electrophysiology recording result also showed that Aβoligomers could inhibit mEPSC and Bryosta-tin-1 could rescue it.The AMPA/NMDA ratio did not change significantly among each group.Western Blot results also showed higher pre-and post-synaptic markers in the present of Bryostatin-1.The mechanisms underlying the protective effect of Bryostatin-1 was in-vestigated.Western Blot results showed that Bryostatin-1 could activate mTOR-S6K1 signal pathway.At last,GFP-RFP-LC3 plasmid was transfected into primary hippocampus neuron culture.Confocolmicroscopy results showed that Aβoligomers could inhibit the au-tophagic flux,and Bryostatin-1could promote the autophagic flux in the present of Aβoligomers.In summary,the present study showed that Bryostatin-1 could activate mTOR-S6K1 signal pathway and then promote autophagic flux in the present of Aβoligomers,which may be the mechanisms underlying the protective effect of PKC against Aβoligomers induced synaptic toxicity.

关 键 词：阿尔茨海默病 蛋白激酶C 神经元树突棘形态 神经元突触功能 神经元自噬通量 

分 类 号：Q421[生物学—神经生物学]