Altered vascular smooth muscle function in the ApoE knockout mouse during the progression of atherosclerosis

- Expression and function of SERCA and PMCA are temporally altered in ApoE−/− VSM.
- TG-induced increases in [Ca2+]c were reduced in ApoE−/− aortic SM cells.
- Ca2+ extrusion is upregulated in isolated ApoE−/− aortic SM cells.

ObjectivesRelaxation of vascular smooth muscle (VSM) requires re-uptake of cytosolic Ca2+ into the sarcoplasmic reticulum (SR) via the Sarco/Endoplasmic Reticulum Ca2+ ATPase (SERCA), or extrusion via the Plasma Membrane Ca2+ ATPase (PMCA) or sodium Ca2+ exchanger (NCX). Peroxynitrite, a reactive species formed in vascular inflammatory diseases, upregulates SERCA activity to induce relaxation but, chronically, can contribute to atherogenesis and altered vascular function by escalating endoplasmic reticulum stress. Our objectives were to determine if peroxynitrite-induced relaxation and Ca2+ handling processes within vascular smooth muscle cells were altered as atherosclerosis develops.MethodsAortae from control and ApoE−/− mice were studied histologically, functionally and for protein expression levels of SERCA and PMCA. Ca2+ responses were assessed in dissociated aortic smooth muscle cells in the presence and absence of extracellular Ca2+.ResultsRelaxation to peroxynitrite was concentration-dependent and endothelium-independent. The abilities of the SERCA blocker thapsigargin and the PMCA inhibitor carboxyeosin to block this relaxation were altered during fat feeding and plaque progression. SERCA levels were progressively reduced, while PMCA expression was upregulated. In ApoE−/− VSM cells, increases in cytosolic Ca2+ [Ca2+]c in response to SERCA blockade were reduced, while SERCA-independent Ca2+ clearance was faster compared to control.ConclusionAs atherosclerosis develops in the ApoE−/− mouse, expression and function of Ca2+ handling proteins are altered. Up-regulation of Ca2+ removal via PMCA may offer a potential compensatory mechanism to help normalise the dysfunctional relaxation observed during disease progression.