Renal macrophage migration and crystal phagocytosis via inflammatory-related gene expression during kidney stone formation and elimination in mice: Detection by association analysis of stone-related gene expression and microstructural observation

Abstract

Mice have a strong ability to eliminate renal calcium oxalate crystals, and our previous examination indicated a susceptibility in which monocyte-macrophage interaction could participate in the phenomenon. To clarify the macrophage-related factors playing roles in the prevention of crystal formation in mouse kidneys, morphologic and expression studies based on microarray pathway analysis were performed. Eight-week-old male C57BL/6N mice were administered 80 mg/kg of glyoxylate by daily intraabdominal injection for 15 days, and the kidneys were extracted every 3 days for DNA microarray analysis. Based on the raw data of microarray analysis, pathway analyses of inflammatory response demonstrated macrophage activation through the increased expression of chemokine (C-X-C) ligand 1, fibronectin 1, and major histocompatability (MHC) class II. Association analysis of related gene expression values by quantitative reverse transcription polymerase chain reaction (RT-PCR) indicated the high association of chemokine (C-C) ligand 2, CD44, colony-stimulating factor 1, fibronectin 1, matrix gla protein, secreted phosphoprotein 1, and transforming growth factor b1 (TGF-b1) with the amount of both renal crystals and F4/80, a macrophage marker. Immunohistochemically, interstitial macrophages increased during the experimental course, and CD44 and MHC class II were upregulated around crystal-formation sites. Ultrastructural observation of renal macrophages by transmission electron microscopy indicated interstitial macrophage migration with the phagocytosis of crystals. In conclusion, increased expression of inflammation-related genes of renal tubular cells induced by crystal formation and deposition could induce monocyte-macrophage migration and phagocytosis via the interaction of CD44 with osteopontin and fibronectin. Such crystalremoving ability of macrophages through phagocytosis and digestion might become a new target for the prevention of stone formation. ß

Correlation between the development of calcium oxalate stones and polymorphisms in the fibronectin gene in the Uighur population of the Xinjiang region of China

“…Tsujihata et al (2000Tsujihata et al ( , 2001Tsujihata et al ( , 2006 reported that Fn significantly inhibited the growth and aggregation of calcium oxalate crystals in vitro; in addition, this gene was believed to impart the ability of forming a biofilm on the surface of calcium oxalate crystals, subsequently inhibiting the calcium oxalate crystal-biofilm combination using renal tubular epithelial cells (Tsujikawa et al, 2007;Tsujihata, 2008). Okada et al (2010), on the other hand, reported that the inhibition of Fn in calcium oxalate crystal-forming animal models induced migration, phagocytosis, and digestion of monocytes and macrophages on calcium oxalate crystals, as a result of the interaction between Fn and CD44.…”

Calcium oxalate monohydrate crystals stimulate monocyte chemoattractant protein-1 and transforming growth factor β1 expression in human renal epithelial cells

“…Recent studies have revealed a novel defense mechanism of the kidney to avoid stone formation, post-adhesion crystal clearing (11)(12)(13). That means the progression of nephrocalcinosis not only depends on the extent, rate and duration of intratubular crystal adhesion, but is additionally countered by a stoneelimination mechanism, which may avoid, slow down or even reverse the progression towards stone formation.…”

“…That means the progression of nephrocalcinosis not only depends on the extent, rate and duration of intratubular crystal adhesion, but is additionally countered by a stoneelimination mechanism, which may avoid, slow down or even reverse the progression towards stone formation. In vivo studies based on microarray analysis have indicated that monocytemacrophage interaction participates in this phenomenon, and that the amount of both renal crystals and macrophages are highly associated with several gene expressions, including MCP-1 and TGFβ1 (11,12). CaOx is the most common type of kidney stone, in which COM is the predominant constituent (16).…”

“…Recently, Okada et al found that mice have a higher tolerance to calcium oxalate (CaOx) stone formation than rats, and the generated calcium stone formation and deposits could be eliminated after several days (11,12). Vervaet et al also reported the similar stone-elimination phenomenon in rats and humans and put forward a new concept, stone-elimination ability, as a factor regulating stone formation (13).…”

“…These studies indicate that the kidney has an additional defense mechanism. Previously, a genome-wide analysis using DNA microarrays indicated the possible association of stone formation with the elimination process by the recruitment of monocytes/macrophages (11). Association analysis demonstrated several genes involved in the stone-elimination process, including chemokine ligand 2 (Ccl2), Cd44, colony-stimulating factor 1 (Csf1), matrix gla protein (Mgp), secreted phosphoprotein 1 (Spp1) and transforming growth factor β1 (Tgfb1) (12).…”