Targeting miR-10 alleviates atherosclerosis by enhancing intestinal ABCA1-mediated cholesterol efflux and repairing intestinal barrier.

BACKGROUND AND AIMS: HDL biogenesis is mainly determined by intestinal and hepatic ABCA1. Inhibiting miR-10b increases macrophage ABCA1 but does not affect intestinal/hepatic ABCA1 or plasma HDL cholesterol in ApoE-/- mice. Given that miR-10a and miR-10b (with similar seed sequences) are comparably expressed in intestine/liver, we hypothesize they redundantly regulate intestinal/hepatic ABCA1 and HDL biogenesis.

METHODS

Primary mouse enterocytes and hepatocytes were treated with antagonist miR-10a, antagonist miR-10b, or their combination, followed by systemic inhibition studies in ApoE-/- and C57BL/6 J mice. ABCA1 expression (qPCR/Western blot) and cholesterol efflux to ApoA-I/HDL were measured. Additionally, atherosclerotic lesions and HDL biogenesis in mice were assessed; ApoE-/- mice were further tested for intestinal barrier (intestinal permeability) and systemic inflammation (gut microbiota, inflammatory responses).

RESULTS

Combined, not individual, miR-10a/b inhibition upregulated ABCA1 and increased cholesterol efflux in primary mouse enterocytes. Consistently, in ApoE-/- and wild-type C57BL/6J mice, combined miR-10 inhibition upregulated intestinal ABCA1, promoted HDL biogenesis, enhanced cholesterol efflux, and ultimately exerted anti-atherosclerotic effects. Concurrently, this dual inhibition reduced intestinal cholesterol accumulation, reshaped gut microbiota, and restored intestinal barrier integrity. Specifically, 16S rRNA sequencing shows that miR-10 inhibition decreased Proteobacteria (phylum level), while it increased Bacteroidetes (phylum level) and enriched Bifidobacterium and Lactobacillus (genus level). These effects collectively suppress systemic inflammation and exert anti-atherosclerotic effects in ApoE-/- mice. Additionally, miR-10 inhibition alleviated systemic inflammation in ApoE-/- mice by suppressing TLR4/NLRP3 inflammasome activation, with this suppression further verified in vitro.

CONCLUSIONS

Inhibition of miR-10 attenuates atherosclerosis through enhanced intestinal ABCA1-mediated cholesterol efflux and barrier integrity, paralleling a favorable shift in gut microbiota composition particularly the enrichment of Bifidobacterium and Lactobacillus. This coordinated intestinal improvement reduces systemic inflammation and ultimately suppresses TLR4/NLRP3 inflammasome activation.