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.
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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
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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).
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YD and CC conceived and designed the experiments. NL performed the experiments. All authors read and approved the final manuscript.
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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).
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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)
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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
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DOI: https://doi.org/10.1007/s10571-023-01378-w