Effects Of Spironolactone On Expression Of MCP-1 Induced By High Glucose Or Aldosterone In Rat Glomerular Mesangial Cells

Objective To investigate the effects of spironolactone (Spi) on expression of monocyte chemoattractant protein-1 (MCP-1) induced by high glucose (HG) or aldosterone (Ald) in rat mesangial cells.Methods①The rat glomerular mesangial cells(RMCs) were cultured in the medium with nomal glucose (5.6 mmol/l, NG) and then divided into five group: NG group, HG group(30 mmol/l, HG), Spi 1 group (Spi 10-7mol/l was added in HG group), Spi 2 group (Spi 10-8mol/l was added in HG group), Spi 3 group (Spi 10-9mol/l was added in HG group). After 48 hours' incubation, the MsC and its supernatant were collected.②Another RMCs were cultured and then divided into five group: NG group, Ald group (Ald 10-7mol/l was added), Spi A group (Spi 10-7mol/l and Ald 10-7mol/l were added), Spi B group (Spi 10-8 mol/l and Ald 10-7 mol/l were added), Spi C group (Spi 10-9 mol/l and Ald 10-7 mol/l were added). RT-PCR was used to measure the expression of Mineralocorticoid receptor (MR) and l1β-hydroxysteroid dehydrogenase (11β-HSD2, an enzyme required for MR ligand specificity) and MCP-1 mRNA.The levels of MCP-1 in the supernatants were determined by enzyme-linked immunosorbant assay(ELISA).Results (1) MR, 11β-HSD2 and MCP-1 mRNA expressions were detectable in RMCs by RT-PCR. (2) After 48h exprosure of RMCs to HG, the levels of MCP-1 mRNA and protein were increased significantly compared to that in NG group [1.44±0.29 vs 1.03±0.22, P<0.01; 194.04±50.62 pg/ml vs 90.03±10.77 pg/ml, P<0.01 ], and the antagonist of MR spironolactone (10-7 mol/l, 10-8 mol/l, 10-9 mol/l) treatment could inhibit these effects (P<0.01). (3) After treatment of 10-7mol/l Ald for 48 hours, the levels of MCP-1 mRNA and protein were 1.84±0.29 and 261.23±58.87 pg/ml, significantly higher than the baseline levels (P<0.01), and spironolactone (10-7 mol/l, 10-8 mol/l) treatment could inhibit the increased MCP-1 expressions stimulated by Ald (P<0.01). And there was no statistical diference in the levels of MCP-1 mRNA expression between Spi A group and NG group(P>0.05).Conclusions Spironolactone can acts by specificly binding with MR on RMCs, and significantly down-regulate the MCP-1 mRNA and protein expressions induced by HG or Ald in RMCs.