Monocyte chemoattractant protein-1 has prosclerotic effects both in a mouse model of experimental diabetes and in vitro in human mesangial cells

Sara Giunti, Gregory H Tesch, Silvia Pinach, Davina J Burt, M E Cooper, Paolo Cavallo-Perin, Giovanni Camussi, Gabriella Gruden

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76 Citations (Scopus)

Abstract

Aims/hypothesis Diabetic nephropathy is characterised by mesangial extracellular matrix accumulation. Monocyte chemoattractant protein-1 (MCP-1), a chemokine promoting monocyte infiltration, is upregulated in the diabetic glomerulus. We performed in vitro and in vivo studies to examine whether MCP-1 may have prosclerotic actions in the setting of diabetes, presumably via its receptor, chemokine (C-C motif) receptor 2 (CCR2), which has been described in mesangial cells. Methods Human mesangial cells were exposed to recombinant human (rh)-MCP-1 (100 ng/ml) for 12, 24 and 48 h and to rh-MCP-1 (10, 100 and 200 ng/ml) for 24 h. Fibronectin, collagen IVand transforming growth factor, beta 1 (TGF- 946;1) protein levels were measured by ELISA and pericellular polymeric fibronectin levels by western blotting. The intracellular mechanisms were investigated using specific inhibitors for CCR2, nuclear factor kappa B (NF- 954;B), p38 mitogen-activated protein kinase and protein kinase C, and an anti-TGF- 946;1 blocking antibody. In both non-diabetic and streptozotocin-induced diabetic mice that were deficient or not in MCP-1, glomerular fibronectin accumulation was examined by immunohistochemistry, while cortical Tgf- 946;1 (also known as Tgfb1) and fibronectin mRNA and protein levels were examined by real-time PCR and western blotting. Results Inmesangial cells,MCP-1 binding to CCR2 induced a 2.5-fold increase in fibronectin protein levels at 24 h followed by a rise in pericellular fibronectin, whereas no changes were seen in collagen IV production. MCP-1-induced fibronectin production was TGF- 946;1- and NF- 954;B-dependent. In diabetic mice, loss ofMCP-1 diminished glomerular fibronectin protein production and both renal cortical Tgf- 946;1 and fibronectin mRNA and protein levels.
Original languageEnglish
Pages (from-to)198 - 207
Number of pages10
JournalDiabetologia
Volume51
Issue number1
Publication statusPublished - 2008

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