Effects Of Advanced Glycation End Products And Aminoguanidine On The Produce Of Extracellular Matrix And Expression Of MMP-2

Objectives:To investigate the effects of advanced glycation end products (AGEs) on the expression of matrix metalloproteinase-2 (MMP-2),the content of typeⅣcollagen and laminin in supernatants of cultured human renal mesangial cells (HRMC). To explore the influence of aminoguanidine (AG) on the MMP-2 expression, the effects of AG on the level of typeⅣcollagen and laminin in supernatants of cultured HRMC.Methods:1.HRMC were incubated with different concentrations of AGE-bovine serum albumin (AGE-BSA) for different time, and exposed to different concentrations of aminoguanidine for 24 hours with or without AGE-BSA(200 mg·l-1). 2.The contents of typeⅣcollagen and laminin in the media were detected by radioimmunoassay. 3.The mRNA and protein expression of MMP-2 in HRMC were measured by RT-PCR and Western blotting respectively.Results:1.AGE-BSA increased the level of typeⅣcollagen and laminin time- and dose- dependently in supernatants of cultured HRMC(P<0.05). 2.Aminoguanidine can reduce AGE-BSA-induced HRMC synthesizes and sectetes typeⅣcollagen and laminin. 3.AGE-BSA decreased the expression of MMP-2 in the level of mRNA and protein time- and dose- dependently (P<0.05). 4.The declined expressions of MMP-2 mRNA and protein in AGE-BSA treated HRMC were increased by exposure to aminoguanidine (P<0.05).Conclusions:1.AGE-BSA increases the content of typeⅣcollagen and laminin in cultured HRMC. These findings suggest that AGEs may induce the extracellular matrix imbalance. 2.AGE-BSA down-regulates the expression of MMP-2 mRNA and protein in HRMC. The data indicates that MMP-2 might mediate the role of AGEs in the development of diabetic nephropathy. 3.AG inhibits the upregulation of typeⅣcollagen and laminin in HRMC by AGE-BSA. The depressed expressions of MMP-2 mRNA and protein in HRMC incubated with AGE-BSA are increased by exposure to AG. These results suggest aminoguanidine might play their role of reno-protection via supression of typeⅣcollagen and laminin and upregulation of MMP-2.