Comparative proteomic analysis of rat aorta in a subtotal nephrectomy model

Abstract

Although accelerated atherosclerosis and arteriosclerosis are the main causes of cardiovascular morbidity and mortality in chronic kidney disease (CKD) patients, the molecular pathogenesis remains largely obscure. Our study of the aortic function in a typical CKD model of subtotal nephrectomy (SNX) rats demonstrated phenotypes that resemble CKD patients with aortic stiffness. The 2-DE analysis of rat aortas followed by MS identified 29 up-regulated and 53 down-regulated proteins in SNX rats. Further Western blot and immunohistochemistry analyses validated the decreased HSP27 and increased milk fat globule epidermal growth factor-8 (MFG-E8) in SNX rats. Functional classification of differential protein profiles using KOGnitor revealed that the two major categories involved in aortic stiffness are posttranslational modification, protein turnover, chaperones (23%) and cytoskeleton (21%). Ingenuity Pathway Analysis highlighted cellular assembly and organization, and cardiovascular system development and function as the two most relevant pathways. Among the identified proteins, the clinical significance of the secreted protein MFG-E8 was confirmed in 50 CKD patients, showing that increased serum MFG-E8 level is positively related to aortic stiffness and renal function impairment. Drug interventions with an inhibitor of the angiotensin converting enzyme, enalapril, in SNX rats improved aortic stiffness and decreased MFG-E8 depositions. Together, our studies provide a repertoire of potential biomarkers related to the aortic stiffness in CKD.

Clinical usefulness of novel prognostic biomarkers in patients on hemodialysis

“…91 In the archetypal subtotal nephrectomy rat model, Lin et al measured aortic pulse wave velocity and assessed the underlying alterations of the aortic proteome. 90 Remarkably, these investigators identified 82 proteins in the aorta that were substantially altered. These proteins included members of the chaperone family, proteins involved in the cytoskeleton/contractile apparatus, extracellular matrix proteins, and proteins implicated in oxidative stress, transport of ions or lipids, and various metabolic pathways.…”

“…Furthermore, angiotensin-convertingenzyme inhibition-one of the most efficacious interventions for prevention of cardiovascular and renal diseases-improved aortic function and reduced MFG-E8 deposition in the aortic medial layer in the subtotal nephrectomy rat model. 90 Studies in patients with coronary artery disease have shown that alterations in the urinary proteomic profile associ ated with coronary heart disease are normalized by long-term angiotensin II blockade. 92 Early-onset cardiovascular disease is a common complica tion in patients with CKD.…”

“…86,87 Clinical samples used in proteome analyses include body fluids (for example, blood, plasma, serum and urine), tissues and dialysis fluid. [88][89][90][91][92][93][94][95][96][97] In view of the proteome's complexity, a separation step is essential before mass spectrometry analysis is performed. This separation step can be performed with a variety of techniques, such as two-dimensional gel electrophoresis, ion-exchange liquid chromatography and capillary electrophoresis.…”

Heat shock proteins in chronic kidney disease

“…On the other hand, Lin et al [22] have shown the decreased expression of Hsp27 in the aortas of rats undergoing subtotal nephrectomy. This deficiency was most evident in the advanced atherosclerotic lesions, thus confirming the potentially cytoprotective role of this HSP and its impact on atherogenesis.…”

Aportaciones de la proteómica al estudio de las enfermedades cardiovasculares

“…También se ha aplicado la tecnología proteómica al estudio de la patología vascular asociada a la enfermedad renal crónica. En este sentido, se han observado cambios en la expresión proteica en la aorta de ratas con insuficiencia renal 16 . La mayoría de estas proteínas alteradas pertenecen a las vías de organización y ensamblaje celular, así como a las de funcionamiento y desarrollo del sistema cardiovascular.…”