益肾化浊方对阿尔茨海默病模型小鼠内化作用的影响

Effect of Yishen Huazhuo Formula on the Internalization of Alzheimer′s Disease Model Mice

目的:观察益肾化浊方对快速老化小鼠亚系8(SAMP8)小鼠谷氨酸受体2(GluR2)内化作用的影响,探讨其改善阿尔茨海默病学习记忆能力的作用机制。方法:采用激光共聚焦、蛋白质印迹法(Western Blotting)和免疫共沉淀检测小鼠海马神经元突触后膜GluR2相对表达量,相关蛋白的表达以及各蛋白间相互结合情况。结果:经益肾化浊方干预后,SAMP8小鼠突触后膜GluR2含量较前增多,降低了谷氨酸受体2L-丝氨酸(GluR2 Ser880)蛋白水平,提高了AMPA受体结合蛋白(ABP)、谷氨酸受体结合蛋白(GRIP)水平,同时促进了GluR2-N-乙基马来酰亚胺敏感因子(NSF)结合,抑制了GluR2-转接蛋白复合物(AP2)结合。结论:益肾化浊方可抑制SAMP8小鼠AMPA受体GluR2内化作用,该作用与减轻GluR2磷酸化和上调内化抑制蛋白GRIP、ABP的表达,促进GluR2-NSF的结合,减少GluR2-AP2的结合有关。

Objective:To observe the effect of Yishen Huazhuo Formula on the internalization of glutamate receptor 2(GluR2)in senescence-accelerated prone 8(SAMP8)mice and explore its mechanism in improving learning and memory abilities in Alzheimer′s disease.Methods:Laser confocal microscopy,Western blotting,and immunoprecipitation were used to measure the relative expression of GluR2 in the postsynaptic membrane of hippocampal neurons,the expression of related proteins,and the interactions between proteins in SAMP8 mice.Results:After intervention with Yishen Huazhuo Formula,the content of GluR2 in the postsynaptic membrane of SAMP8 mice increased,the protein level of GluR2 Ser880 decreased,and the levels of ABP and GRIP proteins increased.Additionally,the binding between GluR2 and N-ethylmaleimide-sensitive factor(NSF)was enhanced,while the binding between GluR2 and adaptor protein complex 2(AP2)was inhibited.Conclusion:Yishen Huazhuo Formula can inhibit the internalization of GluR2 in AMPA receptors of SAMP8 mice.This effect is associated with the reduction of GluR2 phosphorylation,upregulation of expression of internalization inhibitory proteins GRIP and ABP,promotion of GluR2-NSF binding,and reduction of GluR2-AP2 binding.