脑源性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，从而保护小胶质细胞。

Cerebral dopamine neurotrophic factor protects microglia by combining with AKT and by regulating FoxO1/mTOR signaling during neuroinflammation.

机构信息

Department of Spine Surgery, Qilu Hospital of Shandong University, Jinan, 250012, China.

Department of Pharmacy, Qilu Hospital of Shandong University, Jinan, 250012, China.

出版信息

Biomed Pharmacother. 2019 Jan;109:2278-2284. doi: 10.1016/j.biopha.2018.11.028. Epub 2018 Nov 28.

DOI:10.1016/j.biopha.2018.11.028

PMID:30551485

Abstract

BACKGROUND

The 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.

METHODS

After 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.

RESULTS

Laser 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.

CONCLUSIONS

CDNF 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.

摘要

背景

小胶质细胞的激活在神经炎症中起着关键作用。先前的研究表明脑源性神经营养因子 (CDNF) 对神经炎症具有保护作用，但涉及的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，通过该激酶，下游 FoxO1/mTOR 途径与细胞炎症、细胞凋亡、代谢等密切相关。因此，我们研究了 CDNF 是否通过该途径调节神经炎症。

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观察 CDNF 是否与 AKT 共定位。Western blot 检测 AKT 及其下游 FoxO1/mTOR 的表达。ELISA 检测 CDNF 对炎性细胞因子的影响，qRT-PCR 检测 AKT 和 FoxO1/mTOR 的 mRNA 水平。

结果

激光共聚焦和免疫共沉淀实验显著揭示了 AKT 和 CDNF 在小胶质细胞中的相互作用。Western blot 结果表明，CDNF 孵育抑制了 AKT/FoxO1/mTOR 信号的激活。此外，CDNF 明显降低了炎性细胞因子的表达。在 qRT-PCR 中，由于 CDNF 的干预，AKT 和其下游 FoxO1/mTOR 的 mRNA 表达逐渐降低。