NF-κB-Regulated miR-99a Modulates Endothelial Cell Inflammation

Abstract EN

Objective.

The present study was performed to investigate the effects and mechanisms of miR-99a on LPS-induced endothelial cell inflammation, as well as the regulation of NF-κB on miR-99a production.

Methods and Results.

ELISA showed that LPS treatment significantly promoted the secretion of inflammatory factors (TNF-α, IL-6, IL-1β, and MCP-1).

LPS treatment also inhibited miR-99a production and promoted mTOR expression and NF-κB nuclear translocation.

Overexpression of miR-99a suppressed the LPS-induced TNF-α, IL-6, IL-1β, and MCP-1 overproduction, mTOR upregulation, and NF-κB nuclear translocation.

The PROMO software analysis indicated NF-κB binding site in the −1643 to −1652 region of miR-99a promoter.

Dual luciferase reporter analysis, electrophoretic mobility shift assays (EMSA), and chromosome immunoprecipitation (ChIP) assays demonstrated that NF-κB promoted the transcription of miR-99a by binding to the −1643 to −1652 region of miR-99a promoter.

Further studies on HUVECs verified the regulatory effects of NF-κB on miR-99a production.

Conclusion.

MiR-99a inhibited the LPS-induced HUVECs inflammation via inhibition of the mTOR/NF-κB signal.

NF-κB promoted miR-99a production by binding to the −1643 to −1652 region of miR-99a promoter.

Considering the importance of endothelial inflammation on cardiovascular diseases, such as atherosclerosis, our results may provide a new insight into the pathogenesis and therapy of atherosclerosis.