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Objective To observe the dynamic changes of matrix metalloproteinase-2 (MMP-2), tissue inhibitor of metalloproteinase-2 (TIMP-2), membrane-type 1 matrix metalloproteinase (MT1-MMP) and connective tissue growth factor (CTGF) expression in high glucose-stimulated glomerular mesangial cells (GMCs) in rats, and investigate the mechanism of the pathogenesis of diabetic nephropathy. Methods Rat HBZY-1 GMCs were cultured and divided into 3 groups: low concentration (5.5mmol/L) D-glucose (LG) group, high concentration (30mmol/L) D-glucose (HG) group and 24.5mmol/L mannitol plus 5.5mmol/L D-glucose (LG+M) group (served as osmotic pressure control). The mRNA and protein expressions of MMP-2, TIMP-2, MT1-MMP and CTGF were detected with semi-quantitative RT-PCR and Western blotting, and the secreted collagen Ⅳ in supernatants of the GMCs was detected by ELISA after cultured for 24, 48, 72 and 96h. Results Compared with LG group, after exposure to high glucose for 24h, MMP-2 expression was slightly increased (increased by 10%±4%) in GMCs in HG group (P