Abstract
Neuroinflammation is a common characteristic of intracranial infection (ICI), which is associated with the activation of astrocytes and microglia. MiRNAs are involved in the process of neuroinflammation. This study aimed to investigate the potential mechanism by which miR-338-3p negatively modulate the occurrence of neuroinflammation. We here reported that the decreased levels of miR-338-3p were detected using qRT-PCR and the upregulated expression of TNF-α and IL-1β was measured by ELISA in the cerebrospinal fluid (CSF) in patients with ICI. A negative association between miR-338-3p and TNF-α or IL-1β was revealed by Pearson correlation analysis. Sprague–Dawley (SD) rats were injected with LPS (50 μg) into left cerebral ventricule (LCV), following which the increased expression of TNF-α and IL-1β and the reduction of miR-338-3p expression were observed in the corpus callosum (CC). Moreover, the expression of TNF-α and IL-1β in the astrocytes and microglia in the CC of LCV-LPS rats were saliently inhibited by the overexpression of miR-338-3p. In vitro, cultured astrocytes and BV2 cells transfected with mimic-miR-338-3p produced less TNF-α and IL-1β after LPS administration. Direct interaction between miR-338-3p and STAT1 mRNA was validated by biological information analysis and dual luciferase assay. Furthermore, STAT1 pathway was found to be implicated in inhibition of neuroinflammation induced by mimic miR-338-3p in the astrocytes and BV2 cells. Taken together, our results suggest that miR-338-3p suppress the generation of proinflammatory mediators in astrocyte and BV2 cells induced by LPS exposure through the STAT1 signal pathway. MiR-338-3p could act as a potential therapeutic strategy to reduce the neuroinflammatory response.
Graphical Abstract
Diagram describing the cellular and molecular mechanisms associated with LPS-induced neuroinflammation via the miR-338-3p/STAT1 pathway. LPS binds to TLRs on astrocytes or microglia to activate the STAT1 pathway and upregulate the production of pro-inflammatory cytokines. However, miR-338-3p inhibits the expression of STAT1 and reduces the production of inflammatory mediators.
Similar content being viewed by others
Data Availability
The datasets during and/or analyzed during the current study are available from the corresponding author upon reasonable request.
Abbreviations
- ICI:
-
Intracranial infection
- CSF:
-
Cerebrospinal fluid
- PWM:
-
Periventricular white matter
- PWMD:
-
PWM damage
- CC:
-
Corpus callosum
- CNS:
-
Central nervous system
- LPS:
-
Lipopolysaccharide
- LCV:
-
Left cerebral ventricule
- NC:
-
Negative control reagent
- AAV:
-
Adeno associated virus
- STAT1:
-
Signal transducer and activator of transcription 1
- TNF-α:
-
Tumour necrosis factor alpha
- IL-1β:
-
Interleukin-1β
- FBS:
-
Fetal bovine serum
- DMEM:
-
Dulbecco’s modified eagle medium
- PBS:
-
Phosphate-buffer saline
- TBST:
-
Tris-buffered saline Tween-20
- qRT-PCR:
-
Quantitative real-time polymerase chain reaction
- SD rats:
-
Sprague–Dawley rats
- SD:
-
Mean ± standard deviation
- si:
-
Small interfering
- WT:
-
Wild-type
- MUT:
-
Mutant
References
Bai J, Wu L, Chen X, Wang L, Li Q, Zhang Y et al (2018) Suppressor of cytokine signaling-1/STAT1 regulates renal inflammation in mesangial proliferative glomerulonephritis models. Front Immunol 9:1982. https://doi.org/10.3389/fimmu.2018.01982
Burfeind KG, Zhu X, Levasseur PR, Michaelis KA, Norgard MA et al (2018) TRIF is a key inflammatory mediator of acute sickness behavior and cancer cachexia. Brain Behav Immun 73:364–374. https://doi.org/10.1016/j.bbi.2018.05.021
Butturini E, Boriero D, Carcereri DPA, Mariotto S (2019) STAT1 drives M1 microglia activation and neuroinflammation under hypoxia. Arch Biochem Biophys 669:22–30. https://doi.org/10.1016/j.abb.2019.05.011
Chen S, Ye J, Chen X, Shi J, Wu W, Lin W et al (2018) Valproic acid attenuates traumatic spinal cord injury-induced inflammation via STAT1 and NF-kappaB pathway dependent of HDAC3. J Neuroinflamm 15(1):150. https://doi.org/10.1186/s12974-018-1193-6
Chen CY, Chao YM, Lin HF, Chen CJ, Chen CS, Yang JL et al (2020a) miR-195 reduces age-related blood-brain barrier leakage caused by thrombospondin-1-mediated selective autophagy. Aghing Cell 19(11):e13236. https://doi.org/10.1111/acel.13236
Chen YM, He XZ, Wang SM, Xia Y (2020b) Delta-opioid receptors, microRNAs, and neuroinflammation in cerebral ischemia/hypoxia. Front Immunol 11:421. https://doi.org/10.3389/fimmu.2020.00421
Chen C, Wang L, Wang L, Liu Q, Wang C (2021) LncRNA CASC15 promotes cerebral ischemia/reperfusion injury via miR-338-3p/ETS1 axis in acute ischemic stroke. Int J Gen Med 14:6305–6313. https://doi.org/10.2147/IJGM.S323237
Colonna M, Butovsky O (2017) Microglia function in the central nervous system during health and neurodegeneration. Annu Rev Immunol 35:441–468. https://doi.org/10.1146/annurev-immunol-051116-052358
Deng Y, Lu J, Sivakumar V, Ling EA, Kaur C (2008) Amoeboid microglia in the periventricular white matter induce oligodendrocyte damage through expression of proinflammatory cytokines via MAP kinase signaling pathway in hypoxic neonatal rats. Brain Pathol 18(3):387–400. https://doi.org/10.1111/j.1750-3639.2008.00138.x
DiSabato DJ, Quan N, Godbout JP (2016) Neuroinflammation: the devil is in the details. J Neurochem 139(Suppl 2):136–153. https://doi.org/10.1111/jnc.13607
Eikelenboom P, van Exel E, Hoozemans JJ, Veerhuis R, Rozemuller AJ et al (2010) Neuroinflammation—an early event in both the history and pathogenesis of Alzheimer’s disease. Neurodegener Dis 7(1–3):38–41. https://doi.org/10.1159/000283480
Esteller M (2011) Non-coding RNAs in human disease. Nat Rev Genet 12(12):861–874. https://doi.org/10.1038/nrg3074
Fellner L, Irschick R, Schanda K, Reindl M, Klimaschewski L, Poewe W et al (2013) Toll-like receptor 4 is required for alpha-synuclein dependent activation of microglia and astroglia. Glia 61(3):349–360. https://doi.org/10.1002/glia.22437
Feng X, Zhan F, Luo D, Hu J, Wei G, Hua F et al (2021) LncRNA 4344 promotes NLRP3-related neuroinflammation and cognitive impairment by targeting miR-138-5p. Brain Behav Immun 98:283–298. https://doi.org/10.1016/j.bbi.2021.08.230
Gaire BP, Choi JW (2021) Critical roles of lysophospholipid receptors in activation of neuroglia and their neuroinflammatory responses. Int J Mol Sci. https://doi.org/10.3390/ijms22157864
Gorina R, Font-Nieves M, Marquez-Kisinousky L, Santalucia T, Planas AM (2011) Astrocyte TLR4 activation induces a proinflammatory environment through the interplay between MyD88-dependent NFkappaB signaling, MAPK, and Jak1/Stat1 pathways. Glia 59(2):242–255. https://doi.org/10.1002/glia.21094
Guerra MC, Tortorelli LS, Galland F, Da RC, Negri E, Engelke DS et al (2011) Lipopolysaccharide modulates astrocytic S100B secretion: a study in cerebrospinal fluid and astrocyte cultures from rats. J Neuroinflammation 8:128. https://doi.org/10.1186/1742-2094-8-128
He L, Chen Z, Wang J, Feng H (2022) Expression relationship and significance of NEAT1 and miR-27a-3p in serum and cerebrospinal fluid of patients with Alzheimer’s disease. BMC Neurol 22(1):203. https://doi.org/10.1186/s12883-022-02728-9
Heneka MT, Kummer MP, Latz E (2014) Innate immune activation in neurodegenerative disease. Nat Rev Immunol 14(7):463–477. https://doi.org/10.1038/nri3705
Hu Y, He W, Yao D, Dai H (2019) Intrathecal or intraventricular antimicrobial therapy for post-neurosurgical intracranial infection due to multidrug-resistant and extensively drug-resistant Gram-negative bacteria: a systematic review and meta-analysis. Int J Antimicrob Agents 54(5):556–561. https://doi.org/10.1016/j.ijantimicag.2019.08.002
Jiang S et al (2021) Melatonin ameliorates axonal hypomyelination of periventricular white matter by transforming A1 to A2 astrocyte via JAK2/STAT3 pathway in septic neonatal rats. J Inflamm Res 14:5919–5937
Jurjevic I, Miyajima M, Ogino I, Akiba C, Nakajima M, Kondo A et al (2017) Decreased expression of hsa-miR-4274 in cerebrospinal fluid of normal pressure hydrocephalus mimics with parkinsonian syndromes. J Alzheimers Dis 56(1):317–325. https://doi.org/10.3233/JAD-160848
Karve IP, Taylor JM, Crack PJ (2016) The contribution of astrocytes and microglia to traumatic brain injury. Br J Pharmacol 173(4):692–702. https://doi.org/10.1111/bph.13125
Kim JH, Choi DJ, Jeong HK, Kim J, Kim DW, Choi SY et al (2013) DJ-1 facilitates the interaction between STAT1 and its phosphatase, SHP-1, in brain microglia and astrocytes: a novel anti-inflammatory function of DJ-1. Neurobiol Dis 60:1–10. https://doi.org/10.1016/j.nbd.2013.08.007
Kramer S, Haghikia A, Bang C, Scherf K, Pfanne A, Duscha A et al (2019) Elevated levels of miR-181c and miR-633 in the CSF of patients with MS: a validation study. Neurol Neuroimmunol Neuroinflamm 6(6):e623. https://doi.org/10.1212/NXI.0000000000000623
Kwon HS, Koh SH (2020) Neuroinflammation in neurodegenerative disorders: the roles of microglia and astrocytes. Transl Neurodegener 9(1):42. https://doi.org/10.1186/s40035-020-00221-2
Leavitt RJ, Acharya MM, Baulch JE, Limoli CL (2020) Extracellular vesicle-derived miR-124 resolves radiation-induced brain injury. Cancer Res 80(19):4266–4277. https://doi.org/10.1158/0008-5472.CAN-20-1599
Li C, Li X, Zhang Y, Wu L, He J, Jiang N et al (2022a) DSCAM-AS1 promotes cervical carcinoma cell proliferation and invasion via sponging miR-338-3p. Environ Sci Pollut Res Int 29(39):58906–58914. https://doi.org/10.1007/s11356-022-19962-w
Li S, Li L, Li J, Liang X, Song C et al (2022b) miR-203, fine-tunning neuroinflammation by juggling different components of NF-kappaB signaling. J Neuroinflamm 19(1):84. https://doi.org/10.1186/s12974-022-02451-9
Liddelow S, Barres B (2015) SnapShot: astrocytes in health and disease. Cell 162(5):1170. https://doi.org/10.1016/j.cell.2015.08.029
Liu M, Cheng X, Yan H, Chen J, Liu C et al (2021) MiR-135-5p alleviates bone cancer pain by regulating astrocyte-mediated neuroinflammation in spinal cord through JAK2/STAT3 signaling pathway. Mol Neurobiol 58(10):4802–4815. https://doi.org/10.1007/s12035-021-02458-y
Liu Y, Han K, Cao Y, Hu Y, Shao Z, Tong W et al (2022) KLF9 regulates miR-338-3p/NRCAM axis to block the progression of osteosarcoma cells. J Cancer 13(6):2029–2039. https://doi.org/10.7150/jca.63533
Lu S, Wu X, Xin S, Zhang J, Lin H, Miao Y et al (2022) Knockdown of circ_0001679 alleviates lipopolysaccharide-induced MLE-12 lung cell injury by regulating the miR-338-3p/mitogen-activated protein kinase 1 axis. Bioengineered 13(3):5803–5817. https://doi.org/10.1080/21655979.2022.2034564
Qin C, Liu Q, Hu ZW, Zhou LQ, Shang K, Bosco DB et al (2018) Microglial TLR4-dependent autophagy induces ischemic white matter damage via STAT1/6 pathway. Theranostics 8(19):5434–5451. https://doi.org/10.7150/thno.27882
Qu X et al (2021) Levistolide A attenuates Alzheimer’s pathology through activation of the PPARgamma pathway. Neurotherapeutics 18(1):326–339
Rackow AR, Judge JL, Woeller CF, Sime PJ, Kottmann RM (2022) miR-338-3p blocks TGFbeta-induced myofibroblast differentiation through the induction of PTEN. Am J Physiol Lung Cell Mol Physiol 322(3):L385–L400. https://doi.org/10.1152/ajplung.00251.2021
Ransohoff RM (2016) How neuroinflammation contributes to neurodegeneration. Science 353(6301):777–783. https://doi.org/10.1126/science.aag2590
Ravi K, Paidas MJ, Saad A, Jayakumar AR (2021) Astrocytes in rare neurological conditions: morphological and functional considerations. J Comp Neurol 529(10):2676–2705. https://doi.org/10.1002/cne.25118
Salmena L, Poliseno L, Tay Y, Kats L, Pandolfi PP (2011) A ceRNA hypothesis: the Rosetta Stone of a hidden RNA language? Cell 146(3):353–358. https://doi.org/10.1016/j.cell.2011.07.014
Singer BH, Dickson RP, Denstaedt SJ, Newstead MW, Kim K, Falkowski NR et al (2018) Bacterial dissemination to the brain in sepsis. Am J Respir Crit Care Med 197(6):747–756. https://doi.org/10.1164/rccm.201708-1559OC
Sofroniew MV (2015) Astrocyte barriers to neurotoxic inflammation. Nat Rev Neurosci 16(5):249–263. https://doi.org/10.1038/nrn3898
Sun P, Zhang K, Hassan SH, Zhang X, Tang X, Pu H et al (2020) Endothelium-targeted deletion of microRNA-15a/16-1 promotes poststroke angiogenesis and improves long-term neurological recovery. Circ Res 126(8):1040–1057. https://doi.org/10.1161/CIRCRESAHA.119.315886
Sun X, Zhang C, Tao H, Yao S, Wu X (2022) LINC00943 acts as miR-338–3p sponge to promote MPP(+)-induced SK-N-SH cell injury by directly targeting SP1 in Parkinson’s disease. Brain Res 1782:147814. https://doi.org/10.1016/j.brainres.2022.147814
Suthar R, Sankhyan N (2019) Bacterial infections of the central nervous system. Indian J Pediatr 86(1):60–69. https://doi.org/10.1007/s12098-017-2477-z
Tawaratsumida K, Redecke V, Wu R, Kuriakose J, Bouchard JJ, Mittag T et al (2022) A phospho-tyrosine-based signaling module using SPOP, CSK, and LYN controls TLR-induced IRF activity. Sci Adv 8(27):q84. https://doi.org/10.1126/sciadv.abq0084
Teng L, Meng R (2019) Long non-coding RNA MALAT1 promotes acute cerebral infarction through miRNAs-mediated hs-CRP regulation. J Mol Neurosci 69(3):494–504. https://doi.org/10.1007/s12031-019-01384-y
Tiwari PC, Pal R (2017) The potential role of neuroinflammation and transcription factors in Parkinson disease. Dialogues Clin Neurosci 19(1):71–80
Tohidpour A, Morgun AV, Boitsova EB, Malinovskaya NA, Martynova GP, Khilazheva ED et al (2017) Neuroinflammation and infection: molecular mechanisms associated with dysfunction of neurovascular unit. Front Cell Infect Microbiol 7:276. https://doi.org/10.3389/fcimb.2017.00276
Wan L, Jia RM, Ji LL, Qin XM, Hu L, Hu F et al (2022) AMPK-autophagy-mediated inhibition of microRNA-30a-5p alleviates morphine tolerance via SOCS3-dependent neuroinflammation suppression. J Neuroinflamm 19(1):25. https://doi.org/10.1186/s12974-022-02384-3
Xie J, Li S, Ma X, Li R, Zhang H, Li J et al (2022) MiR-217–5p inhibits smog (PM2.5)-induced inflammation and oxidative stress response of mouse lung tissues and macrophages through targeting STAT1. Aging (albany NY) 14(16):6796–6808. https://doi.org/10.18632/aging.204254
Yang X, Zi XH (2019) LncRNA SNHG1 alleviates OGD induced injury in BMEC via miR-338/HIF-1alpha axis. Brain Res 1714:174–181. https://doi.org/10.1016/j.brainres.2018.11.003
Yang Y et al (2021) Vitamin D protects glomerular mesangial cells from high glucose-induced injury by repressing JAK/STAT signaling. Int Urol Nephrol 53(6):1247–1254
Zeng Y, Cullen BR (2004) Structural requirements for pre-microRNA binding and nuclear export by Exportin 5. Nucleic Acids Res 32(16):4776–4785. https://doi.org/10.1093/nar/gkh824
Zhang JX, Lu J, Xie H, Wang DP, Ni HE, Zhu Y et al (2019) circHIPK3 regulates lung fibroblast-to-myofibroblast transition by functioning as a competing endogenous RNA. Cell Death Dis 10(3):182. https://doi.org/10.1038/s41419-019-1430-7
Zhang C, Kang L, Zhu H, Li J, Fang R (2021) MiRNA-338–3p/CAMK IIalpha signaling pathway prevents acetaminophen-induced acute liver inflammation in vivo. Ann Hepatol 21:100191. https://doi.org/10.1016/j.aohep.2020.03.003
Zhao WX, Zhang JH, Cao JB, Wang W, Wang DX, Zhang XY et al (2017) Acetaminophen attenuates lipopolysaccharide-induced cognitive impairment through antioxidant activity. J Neuroinflamm 14(1):17. https://doi.org/10.1186/s12974-016-0781-6
Zhou Q et al (2021) Melatonin reduces neuroinflammation and improves axonal hypomyelination by modulating M1/M2 microglia polarization via JAK2-STAT3-telomerase pathway in postnatal rats exposed to lipopolysaccharide. Mol Neurobiol 58(12):6552–6576
Zhou S, Li J, Zhang X, Xiong W (2022) MicroRNA-124 modulates neuroinflammation in acute methanol poisoning rats via targeting Kruppel-like factor-6. Bioengineered 13(5):13507–13519. https://doi.org/10.1080/21655979.2022.2078549
Zhu Y, Ma C, Lv A, Kou C (2021) Circular RNA circ_0010235 sponges miR-338-3p to play oncogenic role in proliferation, migration and invasion of non-small-cell lung cancer cells through modulating KIF2A. Ann Med 53(1):693–706. https://doi.org/10.1080/07853890.2021.1925736
Zhu J, Zhong F, Chen F, Yang Y, Liao Y, Cao L et al (2022) circRNA_0001679/miR-338-3p/DUSP16 axis aggravates acute lung injury. Open Med (wars) 17(1):403–413. https://doi.org/10.1515/med-2022-0417
Zingale VD, Gugliandolo A, Mazzon E (2021) MiR-155: an important regulator of neuroinflammation. Int J Mol Sci. https://doi.org/10.3390/ijms23010090
Funding
This study was supported by Key Research and Development projects of the Ministry of Sience and Technology (Grant Numbers: 2021YFC2501802), National Natural Science Foundation of China (Grant Numbers: 82272189, 82072230), Natural Science Foundation of Guangdong Province (Grant Number: 2019A1515010206), Guangzhou Municipal Science and Technology Project (Grant Number: 202002030094) and High-level Hospital Construction Project (Grant Number: DFJH201804).
Author information
Authors and Affiliations
Contributions
YD and CC conceived and designed the experiments. NL performed the experiments. All authors read and approved the final manuscript.
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare that they have no competing interests.
Consent to Participate
Informed consent was obtained from all individual participants included in the study.
Consent to Publish
The authors affirm that human research participants provided informed consent for the publication of the images in Table 1.
Ethical Approval
All animals were handled according to the protocols of the Institutional Animal Care and Use Committee, Guangdong Province, China (Animal Certificate No.: SYXK2012-0081). The human CSF specimen was conducted following the Declaration of Helsinki and was approved by the Research Ethics Committee of Guangdong Province’s General Hospital (KY-Q-2022-176-02).
Antibodies
We declared that a limitation in the study was the lack of testing of the specificity of the primary antibodies.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
10571_2023_1378_MOESM1_ESM.tif
Supporting Fig.1 A chart to describe different treatments for astrocyte or BV2 cells in each group. a, b. In the LPS+mimic-NC or LPS+mimic-miR-338-3p group, astrocytes and BV2 cells were first treated with 50nM mimic-NC or mimic-miR-338-3p for 24h, and then astrocytes were treated with LPS for 24h, and BV2 cells were treated with LPS for 6h. c, d. In the LPS+mimic-miR-338-3p+plasmid-STAT1 group, astrocytes and BV2 cells were first transfected with 50nM mimic-miR-338-3p and 0.8μg plasmid-STAT1 for 48h, then astrocytes were administrated with LPS for 24h and BV2 cells were administrated with LPS for 6h. e, f. In the LPS+si-NC or LPS+si-STAT1 group, astrocytes and BV2 cells were first transfected with 50nM si-NC or si-STAT1 for 24h, then astrocytes were administrated with LPS for 24h and BV2 cells were administrated with LPS for 6h. (TIF 3249 KB)
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Liu, N., Zhou, Q., Wang, H. et al. MiRNA-338-3p Inhibits Neuroinflammation in the Corpus Callosum of LCV-LPS Rats Via STAT1 Signal Pathway. Cell Mol Neurobiol 43, 3669–3692 (2023). https://doi.org/10.1007/s10571-023-01378-w
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10571-023-01378-w