Bioinformatics Analysis of Proteomic Profiles During the Process of Anti-Thy1 Nephritis

Abstract

Anti-Thy1 nephritis is a well-established experimental mesangial proliferative nephritis model. Exploring the molecular mechanisms of pathophysiology in anti-Thy1 nephritis may elucidate the pathogeneses of mesangial proliferation. We examined the roles and acting mechanisms of differentially expressed proteins (DEPs) by bioinformatics analysis of glomeruli proteomic profiles during the course of anti-Thy1 nephritis. In total, 108 DEPs were found by two-dimensional fluorescence difference gel electrophoresis (2D-DIGE), and 40 DEPs were identified by matrix-assisted laser desorption ionization/time of flight and liquid chromatography-MS. DEPs were classified into five clusters (Clusters 1-5), according to their expression trends using Cluster 3.0 software, involved in regulating biological processes such as the stress response, cell proliferation, apoptosis, energy metabolism, transport, and the actin cytoskeleton. The expression patterns of ten DEPs, distributed across five clusters, including AKR1A1, AGAT, ATP6V1B2, HIBADH, MDH1, MPST, NIT2, PRDX6, PSMB7, and TPI1, were validated by Western blotting. Based on Western blotting and immunohistochemistry, we also found that the DEP FHL2, which was primarily expressed in the mesangial region, was downregulated on days 3 and 5, and up-regulated on day 10. In vitro, we found that FHL2 overexpression induced mesangial cell proliferation by increasing the number of Sphase cells and decreasing G2/M-phase cells, whereas inhibiting FHL2 had the opposite effect. This study explored novel DEPs and their expression patterns during anti-Thy1 nephritis, and elucidated FHL2's effect on mesangial cell proliferation. These results will contribute to our understanding of the pathogenesis of mesangial proliferation. Molecular & Cellular Proteomics 11: 10.1074/mcp.M111.008755, 1-13, 2012.Mesangial proliferative glomerulonephritis (MesPGN) 1 is the most common chronic kidney disease and is characterized by pathological changes in mesangial cell proliferation and extracellular matrix accumulation. As an important cause of end-stage renal disease, MesPGN has been a major focus of renal disease research. Examining the molecular mechanisms of MesPGN may elucidate its pathogenesis and thereby facilitate the development of clinical treatments. Animal models, such as rat anti-Thy1 nephritis, provide a useful tool to explore these mechanisms.Anti-Thy1 nephritis is a well-established model of mesangial proliferative glomerulonephritis with two major physiological phases (1): the mesangial proliferative phase (e.g. 5 and 7 days after anti-Thy1 antibody injection) and the recovery phase (e.g. 10 and 14 days after anti-Thy1 antibody injection). Biological functions such as stress (2), cytokine action (3, 4), cell proliferation (5, 6), and apoptosis (7, 8) mediate these changes in pathology during the process of anti-Thy1 nephritis. Many proteins have been reported to be involved in regulating these biological functions. For example, Porst (9) reported that fibrillin-1 may regulate mesangial cell ...

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“…Glomerular proteomic profiling revealed that Nit2 is significantly decreased during the course of antiarthritic-Thy1-induced nephritis in adult male Wistar rats (Lu et al 2012). However, the biological significance of these findings is unknown.…”

CXCL10 expression induced by Mxi1 inactivation induces mesangial cell apoptosis in mouse Habu nephritis

“…Traditional anti-Thy-1 nephritis is a classic, reversible model of MesPGN. Certain studies have shown that 2-3 days after antibody injection, rat kidneys demonstrate acute mesangial injury that primarily results in mesangial dissolution and mesangial cell apoptosis [16,17]. The MesPGN model induced by HSV injection (Habu nephritis) is a self-limiting nephritic model that results in acute focal damage and mesangial cell proliferation in rats, rabbits and mice [28][29][30].…”

“…In a previous study regarding the gene expression profiles in an antiThy-1 nephritic model, the results revealed significantly increased expression levels of inflammation-related factors, including the chemokines CCL2, CXCL10 and CXCL13 and the cell adhesion molecules ICAM1 and VCAM1, suggesting that the upregulation of inflammationrelated factors is associated with anti-Thy-1 nephritic pathological processes [16,17]. Because the role of CCL2 has been well documented in human and rat MesPGN, we selected CXCL10 to explore the roles of chemokines in the MesPGN pathological process in mice.…”

“…Although the specific details of the causes and pathogenesis of MesPGN remain unclear, MesPGN is known to be an autoimmune inflammatory disease that involves various pathways; therefore, the causes of this disease are likely related to the immune-mediated inflammatory response [13][14][15]. Based on the microarray results from anti-Thy-1 nephritis models, the expression levels of chemokines, such as CCL2, CCL5 and CCL6, are significantly higher during the early stage of modeling, suggesting that inflammatory-reaction-related cytokines play an important role in the pathological progression of MesPGN [16,17].…”

Applications of Urinary Proteomics in Renal Disease Research Using Animal Models

“…Proliferative mesangial models, including the rat acute anti-Thy1 nephritis model and mouse snake venom-triggered glomerulonephritis, are characterised by initial mesangiolysis, followed by mesangial cell proliferation and accumulation of mesangial matrix, in turn causing proteinuria. The conditions subsequently resolve, and the histology becomes almost normal [9,12]. Membranous nephropathy models, such as the passive Heymann nephritis model (PHN), are characterised by the formation of membrane attack complexes inducing injury to podocytes and expansion of the glomerular basement membrane, in turn causing high-level proteinuria [10,27].…”