Knockdown of NEAT1 induced microglial M2 polarization via miR‑374a‑5p/NFAT5 axis to inhibit inflammatory response caused by OGD/R

Abstract

The long non‑coding RNAs (lncRNAs) have been important regulators for the progression of ischemic‑induced stroke. We aim to study the role of the lncRNA nuclear enriched abundant transcript 1 (NEAT1) in oxygen and glucose deprivation/reoxygenation (OGD/R) treated microglia. OGD/R injury of CHME5 cells was used as an in vitro stroke model. qRT‑PCR analysis was performed\r\nto examine NEAT1, miR‑374a‑5p, nuclear factor of activated T cells 5 (NFAT5) and cytokines. Western blot assay detected protein levels of NFAT5 and microglia markers. The concentration of cytokines was determined by ELISA. Finally, the target relationships among NEAT1, miR‑374a‑5p and NFAT5 were observed by dual luciferase reporter experiments. After OGD/R treatment of CHME5 cells, NEAT1 and NFAT5 were enhanced, while miR‑374a‑5p was decreased. Moreover, knockdown of NEAT1 induced the shifting of OGD/R treated microglia from M1 to M2 and inhibited the inflammatory cytokines in CHME5 cells. Additionally,\r\nNEAT1 directly targeted miR‑374a‑5p while inhibition of miR‑374a‑5p reversed the role of NEAT1 downregulation in OGD/R treated microglia. Furthermore, miR‑374a‑5p directly regulated NFAT5. Interestingly, miR‑374a‑5p also contributed to the transformation of microglia with OGD/R treatment from M1 to M2 and suppressed relative expression levels of inflammatory factors by inhibiting NFAT5 in CHME5 cells. Knockdown of NEAT1 regulated OGD/R injury of CHME5 cells via miR‑374a‑5p/NFAT5 axis to induce the shifting of microglia from M1 to M2 and inhibit inflammatory response, making it a potential target for stroke treatment.