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Abstract
Human mesangial cells grown in either 5 or 25 mM glucose were cultured on type IV collagen which had been previously control-incubated or in vitro glycated. Northern blot analysis revealed that after 3–7 days in culture mesangial cells on glycated type IV collagen expressed ∼25–200% more αl(IV). ∼20–50% less matrix metalloproteinase 2 (MMP-2). and 65–75% more tissue inhibitor of metalloproteinase 1 (TIMP-1). Decreased immunore-activity (∼30–40%) and collagenolytic activity (-10–40%) corresponding to MMP-2 was also detected in media conditioned during the third day of culture on glycated type IV collagen. These effects on cell function were related to the extent of type IV collagen modification and were similar for cells cultured in 5 or 25mM glucose. Elevated glucose (25 vrs 5mM) increased expression of α1(IV) mRNA (∼40–70%) and in conjunction with matrix glycation resulted in detectable levels of MMP-9 message by northern blot although collagenolytic activity corresponding to MMP-9 was not detectable by zymography. We conclude that glucose and matrix glycation may each alter mesangial cell function, perhaps leading to an imbalance in mesangial matrix synthesis and degradation which could contribute to mesangial expansion characteristic of diabetic renal disease.