Fang Min, Zhang Pei, Zhao Yanxin, Jin Aiping, Liu Xueyuan
Department of Neurology, Shanghai Tenth People's Hospital of Tongji University Shanghai, China.
Am J Transl Res. 2016 Aug 15;8(8):3471-81. eCollection 2016.
Sigma receptor is an endoplasmic reticulum protein and belongs to non-opioid receptor. Increasing evidence shows that Sigma receptor activation can significantly attenuate AD induced neurological dysfunction and the functional deficiency of Sigma receptor plays an important role in the Aβ induced neuronal loss. This study aimed to investigate the influence of extracellular accumulation of Aβ on the Sigma receptor expression. Our results showed the increase in extracellular Aβ had little influence on the mRNA expression of Sigma receptor, but gradually reduced its protein expression. Co-immunoprecipitation was employed to evaluate the interaction of Sigma receptor with other proteins. Results showed BIP could bind to Sigma receptor to affect the ubiquitination of Sigma receptor. Further investigation showed there was a NFAT binding site at the promoter of BIP. Then, Western blot assay was performed to detect NFAT expression. Results showed extracellular Aβ affected the nuclear translocation of NFAT and the CaN activity of NFAT also increased with the accumulation of extracellular Aβ. In this study, NFAT-BIP luciferase reporter gene system was constructed. Results showed NFAT was able to regulate the transcription of BIP. Thus, we speculate that extracellular Aβ accumulation may activate CaN/NFAT signaling pathway to induce chaperone BIP expression, which results in Sigma receptor ubiquitination and its degradation.
西格玛受体是一种内质网蛋白,属于非阿片类受体。越来越多的证据表明,西格玛受体激活可显著减轻阿尔茨海默病(AD)诱导的神经功能障碍,且西格玛受体的功能缺陷在淀粉样β蛋白(Aβ)诱导的神经元丢失中起重要作用。本研究旨在探讨细胞外Aβ蓄积对西格玛受体表达的影响。我们的结果表明,细胞外Aβ增加对西格玛受体的mRNA表达影响不大,但逐渐降低其蛋白表达。采用免疫共沉淀法评估西格玛受体与其他蛋白的相互作用。结果显示,结合免疫球蛋白蛋白(BIP)可与西格玛受体结合,影响西格玛受体的泛素化。进一步研究表明,BIP启动子处存在一个活化T细胞核因子(NFAT)结合位点。然后,进行蛋白质免疫印迹分析以检测NFAT表达。结果显示,细胞外Aβ影响NFAT的核转位,且随着细胞外Aβ的蓄积,NFAT的钙调神经磷酸酶(CaN)活性也增加。在本研究中,构建了NFAT-BIP荧光素酶报告基因系统。结果显示,NFAT能够调节BIP的转录。因此,我们推测细胞外Aβ蓄积可能激活CaN/NFAT信号通路,诱导伴侣蛋白BIP表达,从而导致西格玛受体泛素化及其降解。