Cerebral dopamine neurotrophic factor protects microglia by combining with AKT and by regulating FoxO1/mTOR signaling during neuroinflammation
Cerebral dopamine neurotrophic factor protects microglia by combining with AKT and by regulating FoxO1/mTOR signaling during neuroinflammation
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大脑多巴胺神经营养因子通过与 AKT 结合并在神经炎症过程中调节 FoxO1/mTOR 信号传导来保护小胶质细胞
DOI：
10.1016/j.biopha.2018.11.028
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发表时间：
2019
影响因子：
7.5
通讯作者：
Zhao Hua
中科院分区：
文献类型：
--
作者：
Zhang Yayun;Xiang Yanxiao;Wang Xi;Zhu Lin;Li Hao;Wang Songgang;Pan Xin;Zhao Hua
背景小胶质细胞的激活在神经炎症中起着至关重要的作用。既往研究表明脑多巴胺神经营养因子（CDNF）对神经炎症具有保护作用，但其机制尚未得到充分研究。 AKT是一种在哺乳动物中广泛表达的丝氨酸-苏氨酸蛋白激酶，其下游通路FoxO1/mTOR与细胞炎症、凋亡、代谢等密切相关。因此，我们探讨CDNF是否通过该通路调节神经炎症。方法用CDNF和LPS预处理后，通过激光共聚焦显微镜、免疫共沉淀和免疫荧光法检测小胶质细胞，观察CDNF是否与AKT共定位。 Western blot检测AKT及其下游FoxO1/mTOR的表达。采用ELISA法检测CDNF对炎性细胞因子的影响，qRT-PCR检测AKT和FoxO1/mTOR mRNA水平。结果激光共聚焦和免疫共沉淀实验显着揭示了小胶质细胞中AKT和CDNF相互作用的发生。 Western blot 结果显示 CDNF 孵育抑制了 AKT/FoxO1/mTOR 信号传导的激活。此外，CDNF 明显降低炎症细胞因子的表达。 qRT-PCR结果显示，由于CDNF的干预，AKT及其下游FoxO1/mTOR mRNA表达逐渐下降。结论CDNF联合AKT，调节小胶质细胞下游通路FoxO1/mTOR，最终抑制炎症因子的分泌。因此，CDNF可能通过AKT/FoxO1/mTOR信号传导在小胶质细胞的神经炎症中发挥保护作用。
BackgroundThe activation of microglia plays a crucial role in neuroinflammation. Previous studies have shown that cerebral dopamine neurotrophic factor (CDNF) has a protective effect on neuroinflammation, but the mechanisms involved have not been fully studied. AKT is a serine-threonine protein kinase widely expressed in mammals through which the downstream pathway FoxO1/mTOR is closely related to cell inflammation, apoptosis, metabolism, etc. Therefore, we examined whether CDNF regulates neuroinflammation through this pathway.MethodsAfter pretreatment with CDNF and LPS, microglial cells were detected by laser confocal microscopy, coimmunoprecipitation and immunofluorescence to observe whether CDNF was colocalized with AKT. The expression of AKT and its downstream FoxO1/mTOR were determined by Western blot. The effect of CDNF on inflammatory cytokines was detected by ELISA, and the mRNA levels of AKT and FoxO1/mTOR were detected by qRT-PCR.ResultsLaser confocal and coimmunoprecipitation experiments significantly reveal the occurrence of interactions between AKT and CDNF in microglia. Western blot results show that CDNF incubation suppressed the activation of AKT/FoxO1/mTOR signaling. Moreover, CDNF clearly decreased the expression of inflammatory cytokines. In qRT-PCR, the expression of mRNA in AKT and its downstream FoxO1/mTOR gradually decreased due to CDNF intervention.ConclusionsCDNF combined with AKT and regulated the downstream pathway FoxO1/mTOR in microglia, eventually suppressing the secretion of inflammatory factors. Therefore, CDNF might play a protective role in the neuroinflammation of microglia via AKT/FoxO1/mTOR signaling.