MiR-141-3p inhibits vascular smooth muscle cell proliferation and migration via regulating Keap1/Nrf2/HO-1 pathway.

Abstract

miR-141-3p is proven to play a prominent role in various inflammation-related diseases. Nonetheless, little is known concerning the function of miR-141-3p in vascular smooth muscle cells (VSMCs) dysfunction and the underlying mechanism. ApoE knockdown (ApoE-/- ) C57BL/6 mice and human VSMCs were employed to establish atherosclerosis (AS) animal model and cell model, respectively. The expressions of miR-141-3p and Keap1 mRNA were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Enzyme-linked immunosorbent assay (ELISA) was conducted to determine inflammatory cytokines IL-6, IL-β and TNF-α. Cell proliferation, migration and apoptosis were analyzed by BrdU assay, Transwell assay and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, respectively. Luciferase reporter assay was carried out to determine the regulatory relationship between miR-141-3p and Keap1. Additionally, Western blot was used to detect the function of miR-141-3p on the expression levels of Keap1, Nrf2 and HO-1 in VSMCs. miR-141-3p was remarkably down-regulated in both AS animal model and cell model while the expression of Keap1 was elevated. Proliferation and migration of VSMCs were suppressed after miR-141-3p mimics transfection and cell apoptosis was promoted. miR-141-3p also inhibited the expressions of IL-6, IL-β, TNF-α and Keap1 but promoted the expressions of Nrf2 and HO-1. Moreover, the binding site between miR-141-3p and the 3'UTR of Keap1 was confirmed. miR-141-3p is down-regulated during AS, and it can alleviate VSMCs' dysfunction by targeting the Keap1/Nrf2/HO-1 axis.