MiR-200c-3p inhibits LPS-induced M1 polarization of BV2 cells by targeting RIP2

다국어 초록 (Multilingual Abstract)

Background: Microglia are important immune cells, which can be induced by lipopolysaccharide (LPS) into M1 phenotype that express pro-inflammatory cytokines. Some studies have shown that microRNAs play critical roles in microglial activation.
Objective: This study was designed to investigate the role of miR-200c-3p in regulating inflammatory responses of LPS-treated BV2 cells.
Methods: The expression of miR-200c-3p in BV2 cells was detected by real-time PCR. Receptor-interacting protein 2 (RIP2) was predicted as a target gene of miR-200c-3p. Their relationship was verified by dual-luciferase reporter assay. The function of miR-200c-3p and RIP2 in microglial polarization and NF-κB signaling was further evaluated.
Results: LPS treatment reduced miR-200c-3p expression in a dose-dependent and time-dependent manner in BV2 cells. LPS treatment increased the expression of M1 phenotype markers inducible nitric oxide synthase (iNOS) and major histocompatibility complex class (MHC)-II, promoted the release of pro-inflammatory cytokines interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α, and enhanced the nuclear translocation and phosphorylation of nuclear factor-kappaB (NF-κB) p65. Reversely, miR-200c-3p mimics down-regulated the levels of these inflammatory factors. Furthermore, RIP2 was identified to be a direct target of miR-200c-3p. RIP2 knockdown had a similar effect to miR-200c-3p mimics. Overexpression of RIP2 eliminated the inhibitory effect of miR-200c-3p on LPS-induced M1 polarization and NF-κB activation in BV2 cells.
Conclusions: MiR-200c-3p mimics suppressed LPS-induced microglial M1 polarization and NF-κB activation by targeting RIP2. MiR-200c-3p/RIP2 might be a potential therapeutic target for the treatment of neuroinflammation-associated diseases.

참고문헌 (Reference)

1 Liu Q, "miRNA-200c-3p is crucial in acute respiratory distress syndrome" 3 : 17021-, 2017

2 Wang R, "miR-29c-3p inhibits microglial NLRP3 inflammasome activation by targeting NFAT5 in Parkinson’s disease" 25 : 364-374, 2020

3 Meng Z, "miR-200c/PAI-2 promotes the progression of triple negative breast cancer via M1/M2 polarization induction of macrophage" 81 : 106028-, 2020

4 Chuang TD, "miR-200c regulates IL8 expression by targeting IKBKB : a potential mediator of inflammation in leiomyoma pathogenesis" 9 : e95370-, 2014

5 Chen P, "XIST promotes apoptosis and the inflammatory response in CSE-stimulated cells via the miR-200c-3p/EGR3 axis" 21 : 215-, 2021

6 Doyle SL, "Toll-like receptors : from the discovery of NFkappaB to new insights into transcriptional regulations in innate immunity" 72 : 1102-1113, 2006

7 Freeman LC, "The pathogenic role of the inflammasome in neurodegenerative diseases" 136 (136): 29-38, 2016

8 Eren E, "Sulforaphane inhibits lipopolysaccharide-induced inflammation, cytotoxicity, oxidative stress, and miR-155 expression and switches to Mox phenotype through activating extracellular signal-regulated kinase 1/2-nuclear factor erythroid 2-related factor 2/antioxidant response element pathway in murine microglial cells" 9 : 36-, 2018

9 Nimmerjahn A, "Resting microglial cells are highly dynamic surveillants of brain parenchyma in vivo" 308 : 1314-1318, 2005

10 Usluoglu N, "RIP2 mediates LPS-induced p38 and IkappaBalpha signaling including IL-12 p40 expression in human monocyte-derived dendritic cells" 37 : 2317-2325, 2007

11 Jaafar R, "RIP2 enhances cell survival by activation of NF-ĸB in triple negative breast cancer cells" 497 : 115-121, 2018

12 Zheng X, "Propofol attenuates inflammatory response in LPS-activated microglia by regulating the miR-155/SOCS1 pathway" 41 : 11-19, 2018

13 Aryanpour R, "Progesterone therapy induces an M1 to M2 switch in microglia phenotype and suppresses NLRP3 inflammasome in a cuprizone-induced demyelination mouse model" 51 : 131-139, 2017

14 Lu C, "Participation of Rip2 in lipopolysaccharide signaling is independent of its kinase activity" 280 : 16278-16283, 2005

15 Li L, "Overexpression of SIRT1 induced by resveratrol and inhibitor of miR-204 suppresses activation and proliferation of microglia" 56 : 858-867, 2015

16 Zheng Y, "NOD2-RIP2 contributes to the inflammatory responses of mice in vivo to Streptococcus pneumoniae" 671 : 43-49, 2018

17 Wang M, "NOD1/RIP2 signalling enhances the microglia-driven inflammatory response and undergoes crosstalk with inflammatory cytokines to exacerbate brain damage following intracerebral haemorrhage in mice" 17 : 364-, 2020

18 Vilhardt F, "Microglia : phagocyte and glia cell" 37 : 17-21, 2005

19 Lv YN, "MicroRNA-27a negatively modulates the inflammatory response in lipopolysaccharide-stimulated microglia by targeting TLR4 and IRAK4" 37 : 195-210, 2017

20 Hong L, "MicroRNA-200c represses IL-6, IL-8, and CCL-5 expression and enhances osteogenic differentiation" 11 : e0160915-, 2016

21 Filios SR, "Micro-RNA-200 is induced by thioredoxin-interacting protein and regulates Zeb1 protein signaling and beta cell apoptosis" 289 : 36275-36283, 2014

22 Chen L, "Micro-RNA-146a protects against cognitive decline induced by surgical trauma by suppressing hippocampal neuroinflammation in mice" 78 : 188-201, 2019

23 Yao L, "Micro-RNA-124 regulates the expression of MEKK3 in the inflammatory pathogenesis of Parkinson’s disease" 15 : 13-, 2018

24 Lind EF, "Micro-RNA 155 is required for optimal CD8+ T cell responses to acute viral and intracellular bacterial challenges" 190 : 1210-1216, 2013

25 Hu X, "MiR-200c-3p increased HDMEC proliferation through the notch signaling pathway" 246 : 897-905, 2021

26 Yin H, "Knockdown of miR-155 protects microglia against LPS-induced inflammatory injury via targeting RACK1 : a novel research for intracranial infection" 14 : 17-, 2017

27 Zhou Y, "Hepatic NOD2 promotes hepatocarcinogenesis via a RIP2-mediated proinflammatory response and a novel nuclear autophagymediated DNA damage mechanism" 14 : 9-, 2021

28 Liu J, "HMGB3 promotes the proliferation and metastasis of glioblastoma and is negatively regulated by miR-200b-3p and miR-200c-3p" 36 : 357-365, 2018

29 Xiong XY, "Functions and mechanisms of microglia/macrophages in neuroinflammation and neurogenesis after stroke" 142 : 23-44, 2016

30 Jiang Y, "Exosomal miR-200c-3p negatively regulates the migraion and invasion of lipopolysaccharide(LPS)-stimulated colorectal cancer(CRC)" 21 : 48-, 2020

31 Magro AM, "Down-regulation of vinculin upon MK886-induced apoptosis in LN18 glioblastoma cells" 54 : 517-526, 2007

32 Kong H, "Changes in microglial inflammation-related and brain-enriched MicroRNAs expressions in response to in vitro oxygen-glucose deprivation" 39 : 233-243, 2014

33 Chen X-M, "A cellular micro-RNA, let-7i, regulates Toll-like receptor 4 expression and contributes to cholangiocyte immune responses against Cryptosporidium parvum infection" 282 : 28929-28938, 2007