Downregulating ANGPTL3 by miR-144-3p promoted TGF-β1-induced renal interstitial fibrosis via activating PI3K/AKT signaling pathway

Abstract

Despite observations of decreased ANGPTL3 (angiopoietin-like protein 3) levels in tubular atrophy and renal interstitial fibrosis (RIF), its functional implications and regulatory mechanisms in RIF remain unclear. This investigation employed unilateral ureteral obstruction (UUO) mice as in vivo model and human proximal kidney tubuloepithelial HK-2 cells under TGF-β1 treatment as in vitro model to explore RIF. The RIF extent was evaluated using H & E staining and Masson's trichrome staining. There was a significant decrease in ANGPTL3 levels and an increase in miR-144-3p, accompanied by heightened expressions of α-SMA, p-PI3K, p-AKT, Collagen I, and Fibronectin in the UUO mice and HK-2 cells treated with TGF-β1. Enhancing ANGPTL3 expression or suppressing miR-144-3p mitigated TGF-β1-induced cellular apoptosis, inflammation, and PI3K/AKT signaling pathway activation, as evidenced by altered levels of α-SMA, Collagen I, Fibronectin, and associated signaling markers. Using a bioinformatics approach, a miR-144-3p binding site was discovered on the ANGPTL3 mRNA, and this finding was subsequently confirmed through luciferase reporter assay. In HK-2 cells stimulated with TGF-β1, the suppression of ANGPTL3 negated the effects of inhibiting miR-144-3p. Under comparable conditions, the use of LY294002, an inhibitor of the PI3K/AKT pathway, nullified the effects caused by the knockdown of ANGPTL3. Collectively, these findings indicate that miR-144-3p exacerbates RIF through PI3K/AKT pathway activation by targeting ANGPTL3, highlighting a novel potential therapeutic target for RIF management.