Effects of antisense p21 oligodeoxynucleotide on the expression of p21 protein and extracellular matrix of human glomerular mesangial cells in hyperglycemic milieu / 中华肾脏病杂志

ABSTRACT

Objective To investigate the effects of antisense p21 oligodeoxynucleotide (p21 ASODN) on the expression of p21 protein and extracellular matrix in cultured human glomerular mesangial cells (HGMC) under high glucose medium. Methods HGMCs were transfected with 50 nmol/L or 100 nmol/L p21 ASODN or scrambled control oligodeoxynucleotide (SCODN) using lipofectamine 2000. After incubation under normal (5.5 mmol/L) or high (30 mmol/L) glucose media and different times, HGMCs lysates were collected and the expression of p21, fibronectin and laminin was examined by Western blot. The secretion of fibronectin and laminin by HGMCs in supernatants of culture media was also detected with EOSA. Results High glucose media significantly stimulated the expression of p21, increased the syntheses and secretion of fibronectin and laminin in cultured HGMCs in a time-dependent manner. Transfection of HGMCs with p21 ASODN not only decreased p21 protein level caused by high glucose media, but also attenuated the expression and secretion of fibronectin and laminin in supematants of HGMCs lysates and culture media. Meanwhile, SCODN had no significant effects on the expression of p21, fibronectin and laminin. Conclusions High glucose promotes the expression of p21 , fibronectin and laminin in cultured HGMCs. Transfection of HGMCs with p21 ASODN can decrease p21 protein level caused by high glucose media, and attenuate the expression of fibronectin and laminin in supematants of HGMCs lysates and culture media. Modulation of p21 expression in HGMCs may work as an effective way to mitigate the progression of diabetic nephropathy.