FBL Promotes LPS-Induced Neuroinflammation by Activating the NF-κB Signaling Pathway
Zhuoyuan Zhang,1,2,* Dan Liu,1,2,* Rui Lv,1,2,* Haoyan Zhao,1,2,* Tianjing Li,1,2 Yutao Huang,2 Zhicheng Tian,2 Xiangyu Gao,2 Peng Luo,2 Xin Li3 1Biochemistry and Molecular Biology, College of Life Science, Northwest University, Xi’an, 710127, People’s Republic of Chi...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Dove Medical Press
2024-04-01
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| Series: | Journal of Inflammation Research |
| Subjects: | |
| Online Access: | https://www.dovepress.com/fbl-promotes-lps-induced-neuroinflammation-by-activating-the-nf-b-sign-peer-reviewed-fulltext-article-JIR |
| _version_ | 1827287506423382016 |
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| author | Zhang Z Liu D Lv R Zhao H Li T Huang Y Tian Z Gao X Luo P Li X |
| author_facet | Zhang Z Liu D Lv R Zhao H Li T Huang Y Tian Z Gao X Luo P Li X |
| author_sort | Zhang Z |
| collection | DOAJ |
| description | Zhuoyuan Zhang,1,2,* Dan Liu,1,2,* Rui Lv,1,2,* Haoyan Zhao,1,2,* Tianjing Li,1,2 Yutao Huang,2 Zhicheng Tian,2 Xiangyu Gao,2 Peng Luo,2 Xin Li3 1Biochemistry and Molecular Biology, College of Life Science, Northwest University, Xi’an, 710127, People’s Republic of China; 2Department of Neurosurgery, Xijing Hospital, Fourth Military Medical University, Xi’an, 710032, People’s Republic of China; 3Department of Anesthesiology, Xijing Hospital, Fourth Military Medical University, Xi’an, 710032, People’s Republic of China*These authors contributed equally to this workCorrespondence: Peng Luo; Xin Li, Email pengluo@fmmu.edu.cn; li_xin_mail@126.comPurpose: Neuroinflammation occurs in response to central nervous system (CNS) injury, infection, stimulation by toxins, or autoimmunity. We previously analyzed the downstream molecular changes in HT22 cells (mouse hippocampal neurons) upon lipopolysaccharide (LPS) stimulation. We detected elevated expression of Fibrillarin (FBL), a nucleolar methyltransferase, but the associated proinflammatory mechanism was not systematically elucidated. The aim of this study was to investigate the underlying mechanisms by which FBL affects neuroinflammation.Methods: RT-real-time PCR, Western blotting and immunofluorescence were used to assess the mRNA and protein expression of FBL in HT22 cells stimulated with LPS, as well as the cellular localization and fluorescence intensity of FBL. BAY-293 (a son of sevenless homolog 1 (SOS1) inhibitor), SR11302 (an activator protein-1 (AP-1) inhibitor) and KRA-533 (a KRAS agonist) were used to determine the molecular mechanisms underlying the effect of FBL. AP-1 was predicted to be the target protein of FBL by molecular docking analysis, and validation was performed with T-5224 (an AP-1 inhibitor). In addition, the downstream signaling pathways of FBL were identified by transcriptome sequencing and verified by RT-real-time PCR.Results: LPS induced FBL mRNA and protein expression in HT22 cells. In-depth mechanistic studies revealed that when we inhibited c-Fos, AP-1, and SOS1, FBL expression decreased, whereas FBL expression increased when KRAS agonists were used. In addition, the transcript levels of inflammatory genes in the NF-kB signaling pathway (including CD14, MYD88, TNF, TRADD, and NFKB1) were elevated after the overexpression of FBL.Conclusion: LPS induced the expression of FBL in HT22 cells through the RAS/MAPK signaling pathway, and FBL further activated the NF-kB signaling pathway, which promoted the expression of relevant inflammatory genes and the release of cytokines. The present study reveals the mechanism by which FBL promotes neuroinflammation and offers a potential target for the treatment of neuroinflammation.Keywords: FBL, neuroinflammation, LPS, molecular docking, transcriptome sequencing |
| first_indexed | 2024-04-24T11:05:04Z |
| format | Article |
| id | doaj.art-46aa7144a4184202bf76168c6f26b0eb |
| institution | Directory Open Access Journal |
| issn | 1178-7031 |
| language | English |
| last_indexed | 2024-04-24T11:05:04Z |
| publishDate | 2024-04-01 |
| publisher | Dove Medical Press |
| record_format | Article |
| series | Journal of Inflammation Research |
| spelling | doaj.art-46aa7144a4184202bf76168c6f26b0eb2024-04-11T18:05:51ZengDove Medical PressJournal of Inflammation Research1178-70312024-04-01Volume 172217223191891FBL Promotes LPS-Induced Neuroinflammation by Activating the NF-κB Signaling PathwayZhang ZLiu DLv RZhao HLi THuang YTian ZGao XLuo PLi XZhuoyuan Zhang,1,2,* Dan Liu,1,2,* Rui Lv,1,2,* Haoyan Zhao,1,2,* Tianjing Li,1,2 Yutao Huang,2 Zhicheng Tian,2 Xiangyu Gao,2 Peng Luo,2 Xin Li3 1Biochemistry and Molecular Biology, College of Life Science, Northwest University, Xi’an, 710127, People’s Republic of China; 2Department of Neurosurgery, Xijing Hospital, Fourth Military Medical University, Xi’an, 710032, People’s Republic of China; 3Department of Anesthesiology, Xijing Hospital, Fourth Military Medical University, Xi’an, 710032, People’s Republic of China*These authors contributed equally to this workCorrespondence: Peng Luo; Xin Li, Email pengluo@fmmu.edu.cn; li_xin_mail@126.comPurpose: Neuroinflammation occurs in response to central nervous system (CNS) injury, infection, stimulation by toxins, or autoimmunity. We previously analyzed the downstream molecular changes in HT22 cells (mouse hippocampal neurons) upon lipopolysaccharide (LPS) stimulation. We detected elevated expression of Fibrillarin (FBL), a nucleolar methyltransferase, but the associated proinflammatory mechanism was not systematically elucidated. The aim of this study was to investigate the underlying mechanisms by which FBL affects neuroinflammation.Methods: RT-real-time PCR, Western blotting and immunofluorescence were used to assess the mRNA and protein expression of FBL in HT22 cells stimulated with LPS, as well as the cellular localization and fluorescence intensity of FBL. BAY-293 (a son of sevenless homolog 1 (SOS1) inhibitor), SR11302 (an activator protein-1 (AP-1) inhibitor) and KRA-533 (a KRAS agonist) were used to determine the molecular mechanisms underlying the effect of FBL. AP-1 was predicted to be the target protein of FBL by molecular docking analysis, and validation was performed with T-5224 (an AP-1 inhibitor). In addition, the downstream signaling pathways of FBL were identified by transcriptome sequencing and verified by RT-real-time PCR.Results: LPS induced FBL mRNA and protein expression in HT22 cells. In-depth mechanistic studies revealed that when we inhibited c-Fos, AP-1, and SOS1, FBL expression decreased, whereas FBL expression increased when KRAS agonists were used. In addition, the transcript levels of inflammatory genes in the NF-kB signaling pathway (including CD14, MYD88, TNF, TRADD, and NFKB1) were elevated after the overexpression of FBL.Conclusion: LPS induced the expression of FBL in HT22 cells through the RAS/MAPK signaling pathway, and FBL further activated the NF-kB signaling pathway, which promoted the expression of relevant inflammatory genes and the release of cytokines. The present study reveals the mechanism by which FBL promotes neuroinflammation and offers a potential target for the treatment of neuroinflammation.Keywords: FBL, neuroinflammation, LPS, molecular docking, transcriptome sequencinghttps://www.dovepress.com/fbl-promotes-lps-induced-neuroinflammation-by-activating-the-nf-b-sign-peer-reviewed-fulltext-article-JIRfblneuroinflammationlpsmolecular dockingtranscriptome sequencing |
| spellingShingle | Zhang Z Liu D Lv R Zhao H Li T Huang Y Tian Z Gao X Luo P Li X FBL Promotes LPS-Induced Neuroinflammation by Activating the NF-κB Signaling Pathway Journal of Inflammation Research fbl neuroinflammation lps molecular docking transcriptome sequencing |
| title | FBL Promotes LPS-Induced Neuroinflammation by Activating the NF-κB Signaling Pathway |
| title_full | FBL Promotes LPS-Induced Neuroinflammation by Activating the NF-κB Signaling Pathway |
| title_fullStr | FBL Promotes LPS-Induced Neuroinflammation by Activating the NF-κB Signaling Pathway |
| title_full_unstemmed | FBL Promotes LPS-Induced Neuroinflammation by Activating the NF-κB Signaling Pathway |
| title_short | FBL Promotes LPS-Induced Neuroinflammation by Activating the NF-κB Signaling Pathway |
| title_sort | fbl promotes lps induced neuroinflammation by activating the nf kappa b signaling pathway |
| topic | fbl neuroinflammation lps molecular docking transcriptome sequencing |
| url | https://www.dovepress.com/fbl-promotes-lps-induced-neuroinflammation-by-activating-the-nf-b-sign-peer-reviewed-fulltext-article-JIR |
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