Purpose-Atherosclerosis' connected diseases are the leading cause of death in Europe, accounting for over two million deaths each year. A high-cholesterol diet is an important factor in the development of atherosclerosis, and ischemic heart disease. In the present study an animal model was exploited in order to obtain insights on plaque formation and its evolution towards instability and rupture. For this purpose, proteins which are actively synthesized and secreted (secretome) by coronaries isolated from pig subjected to Hypercholestorolemic diet, were identified and the expression in the artery wall of some of the most interesting factors were characterised. Methods-Coronaries isolated from pig on standard or Hypercholestorolemic diets were incubated for 24 h in serum-free medium. Released proteins were digested with trypsin. Peptide mixtures were fractioned by nano-HPLC and analyzed by mass spectrometry. Proteins reported as differentially modulated resulted statistical validated across 6 replicates (T-test >95%; pvalue <0.05). Results-With our proteomics investigation we were able to identify 74 released proteins and/or degradation products of cellular proteins. Of these, 13 proteins resulted differentially modulated between control and high cholesterol diet. Several modulated proteins are intracellular and among some involved in the adhesion pathway. Even more interesting are the extracellular matrix components resulted over-expressed in Hypercholestorolemic diet samples. Conclusions-A proteomic approach enabled us to identify factors released by coronary arteries characterized by early plaques induced by Hypercholestorolemic diet. Our experimental strategy intends to disclose molecular components involved in atherosclerosis onset and development that are released by tissues into body fluids and therefore that can be easily measurable in blood for diagnosis and prognosis. This may represent a useful strategy in order to identify molecular pathways activated in early events of atherosclerosis.
Secretome modulation by high-cholesterol diet in pig coronary arteries
CECCHETTINI, ANTONELLA;
2012-01-01
Abstract
Purpose-Atherosclerosis' connected diseases are the leading cause of death in Europe, accounting for over two million deaths each year. A high-cholesterol diet is an important factor in the development of atherosclerosis, and ischemic heart disease. In the present study an animal model was exploited in order to obtain insights on plaque formation and its evolution towards instability and rupture. For this purpose, proteins which are actively synthesized and secreted (secretome) by coronaries isolated from pig subjected to Hypercholestorolemic diet, were identified and the expression in the artery wall of some of the most interesting factors were characterised. Methods-Coronaries isolated from pig on standard or Hypercholestorolemic diets were incubated for 24 h in serum-free medium. Released proteins were digested with trypsin. Peptide mixtures were fractioned by nano-HPLC and analyzed by mass spectrometry. Proteins reported as differentially modulated resulted statistical validated across 6 replicates (T-test >95%; pvalue <0.05). Results-With our proteomics investigation we were able to identify 74 released proteins and/or degradation products of cellular proteins. Of these, 13 proteins resulted differentially modulated between control and high cholesterol diet. Several modulated proteins are intracellular and among some involved in the adhesion pathway. Even more interesting are the extracellular matrix components resulted over-expressed in Hypercholestorolemic diet samples. Conclusions-A proteomic approach enabled us to identify factors released by coronary arteries characterized by early plaques induced by Hypercholestorolemic diet. Our experimental strategy intends to disclose molecular components involved in atherosclerosis onset and development that are released by tissues into body fluids and therefore that can be easily measurable in blood for diagnosis and prognosis. This may represent a useful strategy in order to identify molecular pathways activated in early events of atherosclerosis.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
