Differential genomic changes caused by cholesterol- and PUFA-rich diets in regenerated porcine coronary endothelial cells

Abstract

Endothelial regeneration and dyslipidemia impair endothelium-dependent relaxation, while supplementation with fish oil (FO) prevents it. The genomic impact of different diets was compared in primary cultures derived from native and regenerated endothelial cells. Pigs were fed with high-cholesterol (CHL) or FO-rich diet. Partial in vivo removal of endothelium was performed to induce endothelial regeneration. Native and regenerated cells were harvested, cultured, and prepared for genomic (microarray experiments, real-time PCR) and proteomic (Western blotting) analysis. The analysis identified genomic changes induced by chronic CHL diet in native cultures resembling those induced by in vivo regeneration, as well as those that could be prevented by FO diet. At the protein level, the reduced and increased presences of endothelial nitric oxide synthase and F2, respectively, observed after regeneration combined with CHL diet were alleviated by FO. The comparison of the differential changes induced by regeneration in vivo in endothelial cells from both diet groups revealed a limited number of genes as the most likely contributors to reduction in endothelium-dependent relaxations in porcine coronary arteries lined with regenerated endothelium.

Cyclin-Dependent Kinase 5–Mediated Hyperphosphorylation of Sirtuin-1 Contributes to the Development of Endothelial Senescence and Atherosclerosis

“…In P4 cells, the ratio of phosphorylated SIRT1 (p-SIRT1) to total SIRT1 (t-SIRT1) was augmented Ͼ9-fold compared with that in P1 cells. The p-SIRT1 levels in regenerated endothelial cells collected from porcine aortas 4 weeks after balloon denudation injury 29 were also elevated compared with native cells ( Figure 1A, right). The association of p-SIRT1 with cellular senescence was further confirmed in PAECs treated with hypercholesterolemic serum collected from ApoE Ϫ/Ϫ mice, H 2 O 2 , or low-density lipoprotein cholesterol ( Figure 1B and 1C).…”

“…Native and regenerated endothelial cells were collected from pigs subjected to coronary angioplasty as described. [27][28][29] …”

Neoatherosclerosis: Another Consequence of Endothelial Dysfunction?

“…This experiment identified 5 genes with atheroprotective functions that were downregulated in the regenerated cells while another 9 pro-atherogenic genes were upregulated. 19 Information derived from this and other deep phenotyping studies could be used further to identify genes or proteins that could be targeted for genetic manipulation to restore normal endothelial function. In addition, profiling studies should take into account the effects of antiproliferative drugs eluted from stents on the endothelium.…”

MMP‐7 and Cardiovascular Disease: Not So Surprising!

“…The reason why this proposal is not too surprising, at least to us, is that a few years ago, we studied (with microarrays) the genomic profile of primary cell cultures of native and regenerated (after balloon angioplasty) porcine coronary endothelial cells from the same hearts [2]. More recently, we have repeated those studies in pigs fed chronically with diets rich in either cholesterol or x 3 -unsaturated fatty acids [3]. As always in such scientific fishing expeditions, a large number of genes were either up-or down-regulated.…”

“…However, regenerated endothelial cells expressed the mRNA for two genes that were not found in native cell cultures, MMP-7 and FABP4 [2]. The over-expression persisted in regenerated endothelial cells irrespective of the diet to which the donor animals were subjected [3]. We concluded that the over-expressions of these two genes, MMP7 and FABP4, are major inducing factors of endothelial dysfunction.…”