Andrographolide inhibits murine embryonic neuronal development through PFKFB3-mediated glycolytic pathway

• Published in: European journal of pharmacology Vol. 940; p. 175474
• Main Authors: Shi, Liangqin, Li, Hongfei, Wang, Zheng, Liu, Weiming, Wu, Xiao, Li, Jiali, Jin, Zhong, Hu, Wangming, Guo, Qiru, Zhang, Lingling, Wang, Yang, Liang, Bing, Zhu, Song, Zhang, Haidi, Xu, Huan, Yang, Lan, Shi, Riyi, Wang, Yong
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 05.02.2023
• Subjects: Andrographolide; Animals; Cell Proliferation; Diterpenes - pharmacology; Embryonic neuronal development; Female; Glycolysis; Mice; NeuN; PFKFB3; Phosphofructokinase-2 - genetics; Phosphofructokinase-2 - metabolism; Phosphoric Monoester Hydrolases - metabolism; Pregnancy; TBR1; Glycolysis; TBR1; PFKFB3; NeuN; Andrographolide; Embryonic neuronal development
• ISSN: 0014-2999; 1879-0712
• DOI: 10.1016/j.ejphar.2022.175474

Dysregulation of neuronal development may cause neurodevelopmental disorders. However, how to regulate embryonic neuronal development and whether this regulation can be medical interrupted are largely unknown. This study aimed to investigate whether and how andrographolide (ANP) regulates embryonic neuronal development. The pregnant mice at embryonic day 10.5 (E10.5) were administrated with ANP, and the embryonic brains were harvested at E17.5 or E18.5. Immunofluorescence (IF), Immunohistochemistry (IHC) performed to determine whether ANP is critical in regulating neuronal development. Real-time quantitative PCR, western blotting, cell counting kit-8 assay, Flow Cytometry assay, Boyden Chamber Migration assay carried out to evaluate whether ANP regulates neuronal proliferation and migration. Protein-protein interaction, CO-immunoprecipitation and IF staining carried out to evaluate whether ANP regulates the interaction between PFKFB3, NeuN and TBR1. Knockdown or overexpression of PFKFB3 by adenovirus infection were used to determine whether ANP inhibits neuronal development through PFKFB3 mediated glycolytic pathway. Our data indicated that ANP inhibited the maturation of embryonic neurons characterized by suppressing neuronal proliferation and migration. ANP regulated the interaction between PFKFB3, NeuN, and TBR1. Knockdown of PFKFB3 aggravated ANP mediated inhibition of neuronal proliferation and migration, while overexpression of PFKFB3 attenuated ANP mediated neuronal developmental suppression. In summary, ANP suppressed the expression of PFKFB3, and interrupted the interaction between TRB1 and NeuN, resulting in suppressing neuronal proliferation, migration and maturation and eventually inhibiting murine embryonic neuronal development.