Knockdown of Long Non-coding RNA TUG1 Suppresses pyroptosis in High Glucose-Stimulated BV-2 Cells

Abstract

Abstract Neuroinflammation is a mechanism by which obesity leads to cognitive impairment. LncRNA-TUG1 can increase neuroinflammation by regulating microglial cells activation, but its mechanism is unclear. The purpose of this study was to investigate the potential functional role of long non-coding RNA TUG1 in high glucose -stimulated BV-2 microglia cells. Mouse microglia cell line BV-2 cells were cultured in vitro and TUG1 shRNA was used to knock down its level. After microglial cells were exposed to high glucose 24 h, TUG1 level, as well as some pyroptosis-associated proteins and releasing inflammatory cytokines were detected by quantitative real-time PCR (qRT-PCR), western blot. The activation of gasdermin D (GSDMD) were detected by immunofluorescence staining, qRT-PCR, and western blot. Our results showed that compared with those in the 5.5 mmol/L glucose control (NC) group, expression of TUG1, NLRP3, caspase1, GSDMD, IL-18 and IL-1β in the 25mmol/L glucose (HG) group of cultured BV-2 cells were significantly increased. the expression of TUG1 was up-regulated in microgolial cells 24 h after high glucose treatment. However, the lncRNA-TUG1 knocked down could significantly reduce the expression of NLRP3, caspase-1 and GSDMD protein and mRNA level in BV-2 cells. Besides, immunofluorescence results showed the lncRNA-TUG1 knocked down stained positively for GSDMD. LncRNA-TUG1 knocked could improve microglial pyroptosis and production of inflammatory cytokines after high glucose insult.