Ozone rectal insufflation inhibits the development of atherosclerosis in ApoE-/-mice, which is mediated by the regulation of gut microbiota and metabolites.

Introduction: Atherosclerotic cardiovascular disease is the world's leading cause of death. Researches have proven that ozone therapy can effectively inhibit the development of atherosclerosis; however, the underlying mechanisms remain unclear. This study aims to explore how ozone rectal insufflation (O3-RI) prevents atherosclerosis. Methods: O3-RI therapy involves administering medical ozone via rectal injection to prevent multiple diseases. ApoE-/- mice were fed a high-fat diet (HFD) to induce atherosclerosis. Gut microbiota was identified by 16S rRNA sequencing. Microbial metabolites were determined by liquid chromatography-mass spectrometry(LC-MS). Results: O3-RI reduced serum LDL-C levels by approximately 29.6% and decreased both atherosclerotic lipid areas and plaque area in ApoE-/- mice. Additionally, O3-RI improved gut microbiota imbalance caused by a high-fat diet in these mice. Notably, O3-RI increased beneficial microorganisms like Discussion: This research indicates that O3-RI effectively inhibits atherosclerosis and reduces serum LDL-C levels. These effects are likely linked to ozone's optimization of gut microbiota and regulation of microbial metabolites in ApoE-/- mice. The non-invasive O3-RI may play a crucial role in managing and treating atherosclerosis. The non-invasive O3-RI may play a crucial role in the managing and treating of atherosclerosis.