miR-194-5p inhibits LPS-induced astrocytes activation by directly targeting neurexophilin 1.

Molecular and Cellular Biochemistry
Ming WangQiang Zuo

Abstract

Astrocytes are vitally involved in the development of neurodegenerative diseases and brain cancers. In this work, we investigated the potential ameliorative role of microRNA-194-5p (miR-194-5p) against lipopolysaccharide (LPS)-induced astrocytes activation and the mechanism underneath. Astrocytes were transfected with miR-194-5p mimic or inhibitor and subsequently induced with LPS. Cell proliferation was measured using MTT assay while Transwell assay was used for assessing cell migration. The concentrations of cyclooxygenase 2 (COX2) and cytokines (tumor necrosis factor-α (TNF-α), transforming growth factor β (TGF-β), interleukin (IL)-1β and IL-6) were determined by enzyme-linked immunosorbent assay (ELISA). Gene expression was assessed by quantitative reverse transcription PCR (RT-qPCR) while western blotting was used for quantifying relative protein expression. We found that miR-194-5p, downregulated in LPS-induced astrocytes, significantly inhibited LPS-induced cell proliferation and migration. In addition, miR-194-5p inhibited the release of COX2 and pro-inflammatory cytokines (TNF-α, TGF-β, IL-1β and IL-6). Moreover, the silencing of neurexophilin 1 (NXPH1), an in silico and mechanistically confirmed direct target of miR-1...Continue Reading

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Citations

Aug 8, 2021·International Journal of Molecular Sciences·Hisham KashifSangeetha Sukumari-Ramesh

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Methods Mentioned

BETA
PCR
transfection
electrophoresis
ELISA

Software Mentioned

Targetscan
GraphPad
Image J
GraphPad prism
Quantity One

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