MicroRNA-103 Protects Coronary Artery Endothelial Cells against H₂O₂-Induced Oxidative Stress via BNIP3-Mediated End-Stage Autophagy and Antipyroptosis Pathways

Abstract

Endothelial cell damage caused by oxidative stress is widely considered to be a triggering event in atherosclerosis (AS). However, the specific effect elicited by autophagy in endothelial cells undergoing oxidative stress remains controversial, especially during end-stage autophagy. The inhibition of end-stage autophagy has been reported to increase cell pyroptosis and contribute to endothelial damage. Several studies have shown that microRNA-103 is involved in end-stage autophagy; however, its specific mechanism of action is not yet characterized. In this study, we addressed the regulatory role of miR-103 in autophagy during oxidative stress of endothelial cells. Hydrogen peroxide (H₂O₂) treatment was used as an <i>in vitro</i> model of oxidative stress. MTS and ROS levels were measured to evaluate cell activity. qRT-PCR was used to detect the expression of miR-103. Autophagy was examined using western blot, immunofluorescence staining, and electron microscopy, while western blot analysis detected pyroptosis-related proteins. Results show that miR-103 expression decreased under oxidative stress. Further, miR-103 repressed transcription of Bcl-2/adenovirus E1B 19 kDa interacting protein (BNIP3). The oxidative stress caused by H₂O₂ caused cell damage from 2 hours (<i>P</i> < 0.05) and increased the level of intracellular reactive oxygen species (<i>P</i> < 0.05); at the same time, the damage could be further aggravated by the stimulation of bafA1 (<i>P</i> < 0.05). Under the stimulation of H₂O₂, the expression of miR-103 decreased (<i>P</i> < 0.05). However, high expression of miR-103 could reduce the accumulation of LC3II and P62 (<i>P</i> < 0.05) by inhibiting the downstream target gene Bcl-2/adenovirus E1B 19 kDa interacting protein (BNIP3), thus reducing the occurrence of cell pyroptosis (<i>P</i> < 0.05). This process could be blocked by end-stage autophagy inhibitor bafA1 (<i>P</i> < 0.05), which further indicated that miR-103 affected cell injury by autophagy. On the contrary, the low expression of miR-103 promoted the accumulation of autophagy protein and increased the occurrence of pyroptosis (<i>P</i> < 0.05). In conclusion, inhibition of miR-103 restrained end-stage of autophagy by regulating BNIP3, thus changing the occurrence of cell pyroptosis.