Deeper insight into chronic kidney disease-related atherosclerosis: comparative proteomic studies of blood plasma using 2DE and mass spectrometry

Abstract

BackgroundAtherosclerosis is a major cause of cardiac events and mortality in patients suffering from chronic kidney disease (CKD). Moreover, the risk of cardiovascular disease (CVD) development in patients with CKD increases as kidney function declines. Although the close connection between atherosclerosis and kidney dysfunction is undeniable, particular risk factors and specific mechanisms that promote CVD in patients with CKD remain unclear. To gain insight into better recognition of the mechanisms of accelerated atherosclerosis in patients with CKD, we performed a comparative proteomic analysis of blood plasma from patients in various stages of CKD and thus distinct progression of atherosclerosis (n = 90), patients with advanced CVD and normal renal function (n = 30) and healthy volunteers (n = 30).MethodsPlasma samples were depleted using affinity chromatography and divided into three fractions: high-abundant, low-abundant and low-molecular weight proteins. The first two fractions were analyzed by two-dimensional gel electrophoresis and mass spectrometry, the last one has been subjected to direct MS/MS analysis. A proteomic profiles for high-abundant, low-abundant and low-molecular weight proteins fractions were obtained. Differential accumulated proteins were confirmed by selected reaction monitoring analysis (SRM). The Gene Ontology (GO) function and the interaction networks of differentially expressed proteins were then analyzed.ResultsForty-nine proteins (13 high- and 36 low-molecular mass) showed differences in accumulation levels. For eleven of them differential expression were confirmed by selected reaction monitoring analysis. Bioinformatic analysis showed that identified differential proteins were related to three different processes: the blood coagulation cascade, the transport, binding and metabolism of lipoproteins and inflammatory processes.ConclusionsObtained data provide an additional line of evidence that different molecular mechanisms are involved in the development of CKD- and CVD-related atherosclerosis. The abundance of some anti-atherogenic factors revealed in patients with CKD suggests that these factors are not associated with the reduction of atherosclerosis progression in CKD that is typically observed in “classical” CVD. Moreover, obtained data also suggest that mechanism of CVD acceleration may be different in initial and advanced stages of CKD. Undoubtedly, in advanced stages of CKD inflammation is highly pronounced.Electronic supplementary materialThe online version of this article (doi:10.1186/s12967-014-0378-8) contains supplementary material, which is available to authorized users.

Proteomic Analysis of Human Serum from Patients with Chronic Kidney Disease

“…Our results correlate well with the data obtained by other researchers in that blood APOA4, AAT, VIL1, complement component and cytokine concentrations increased in patients with renal disorders [8,[11][12][13][14]28,[32][33][34]43,[69][70][71]. Moreover, we found elevated serum concentrations of potential oncomarkers (CUL5, antigen KI-67) and other intracellular proteins (NKRF, CAPRI, IGSF22, APPBP2, CCD171 and CCD43) in patients with CKD.…”

“…The disadvantage of this method is its poor effectiveness in diagnosing early CKD. There are a number of studies where potential protein biomarkers of CKD were proposed and evaluated in the serum or plasma of patients [7,8,[11][12][13][14]. Peptide biomarkers can also provide insight into CKD diagnosis and progression.…”

iTRAQ-based proteomic analysis of plasma reveals abnormalities in lipid metabolism proteins in chronic kidney disease-related atherosclerosis

“…We have also demonstrated that apoAIV displays an anti-atherogenic effect only in the case of classical CVD and is not efficient in CKD-A. In that work due to limited resolution of 2DE, apart from apoAIV, two additional proteins related to lipid metabolism (apoB and apoAI) were identified as differentially expressed15. In current, high throughput study, we discovered that the majority of apolipoprotein classes and other proteins involved in lipid metabolism revealed a completely distinct profile in CKD-A compared with CVD.…”

Multiplexed MRM-based protein quantification of putative prognostic biomarkers for chronic kidney disease progression in plasma

“…In addition, several shotgun proteomic studies followed by verification via different assays (Western blot, MRM in small sample sizes (n ≤ 11), or ELISA), supported an upregulation of AMPB in patients of various CKD stages compared to healthy controls, in adipose tissue, urine as well as urine exosomes [74][75][76] . AMBP was also found to progressively increase in plasma of individuals with increasing CKD stage -from CKD1-2 to CKD3-4 and finally to CKD5-based on relative quantification by LC-MS/MS 77,78 or MRM 77 . www.nature.com/scientificreports www.nature.com/scientificreports/ AMBP, SERPINF1, B2M, and HBB have been investigated as HDL (High-density lipoprotein) associated proteins by LC-MS/MS, and were found to correlate with severe kidney damage 79 .…”