Coronary Microvascular Dysfunction Is Associated With Augmented Lysosomal Signaling in Hypercholesterolemic Mice

Abstract

Background Accumulating evidence indicates that coronary microvascular dysfunction (CMD) caused by hypercholesterolemia can lead to myocardial ischemia, with or without obstructive atherosclerotic coronary artery disease. However, the molecular pathways associated with compromised coronary microvascular function before the development of myocardial ischemic injury remain poorly defined. In this study, we investigated the effects of hypercholesterolemia on the function and integrity of the coronary microcirculation in mice and the underlying mechanisms. Methods and Results Mice were fed a hypercholesterolemic Paigen's diet for 8 weeks. Echocardiography data showed that Paigen's diet caused CMD, characterized by significant reductions in coronary blood flow and coronary flow reserve, but did not affect cardiac remodeling or dysfunction. Immunofluorescence studies revealed that Paigen's diet–induced CMD was associated with activation of coronary arterioles inflammation and increased myocardial inflammatory cell infiltration. These pathological changes occurred in parallel with the upregulation of lysosomal signaling pathways in endothelial cells (ECs). Treating hypercholesterolemic mice with the cholesterol‐lowering drug ezetimibe significantly ameliorated Paigen's diet–induced adverse effects, including hypercholesterolemia, steatohepatitis, reduced coronary flow reserve, coronary endothelial cell inflammation, and myocardial inflammatory cell infiltration. In cultured mouse cardiac ECs, 7‐ketocholesterol increased mitochondrial reactive oxygen species and inflammatory responses. Meanwhile, 7‐ketocholesterol induced the activation of transcriptional factor EB and lysosomal signaling in mouse cardiac ECs, whereas the lysosome inhibitor bafilomycin A1 blocked 7‐ketocholesterol–induced transcriptional factor EB activation and exacerbated 7‐ketocholesterol–induced inflammation and cell death. Interestingly, ezetimibe synergistically enhanced 7‐ketocholesterol–induced transcriptional factor EB activation and attenuated 7‐ketocholesterol–induced mitochondrial reactive oxygen species and inflammatory responses in mouse cardiac ECs. Conclusions These results suggest that CMD can develop and precede detectable cardiac functional or structural changes in the setting of hypercholesterolemia and that upregulation of transcriptional factor EB–mediated lysosomal signaling in endothelial cells plays a protective role against CMD.

Keywords

• coronary flow reserve
• coronary microvascular dysfunction
• endothelial dysfunction
• hypercholesterolemia
• lysosome