Treatment with antisense Smad2 oligodeoxynudeotides reduces extracellular matrix secretion in rat mesangial cells cultured with high glucose

Objective To investigate the effects of antisense Smad2 oligodeoxynudeotides(ODN) on fibronetin(FN) and collagen Ⅳ(ColⅣ) secretion of rat mesangial cells cultured with high glucose, explore the action of Smad2 in the glomerulosclerosis and to find a new method to retard the progress of glomerular fibrosis. Methods 20-mer antisense, sense and random ODNs were designed and synthesized that were phosphorothioate modified to increase stability. The antisense ODN encompassed the ATG of the rat Smad2 gene. ODN was tranferred transiently into rat mesangial cells through liposome. Rat cells were treated with high glucose. mRNA and protein of Smad2 were detected by RT-PCR and cytochemistry. FN and ColⅣ were examined by ELISA. Results Antisense ODN significantly decreased mRNA and protein expression of Smad2 in rat mesangial cells treated with high glucose(P 0.05). FN concentrations in the culture medium of high glucose group, antisense group, sense group and random group were (84.19±6.81) ng/ml, (57.65±5.05) ng/ml, (78.72±9.13) ng/ml and (77.13±7.36) ng/ml respectively. ColⅣ concentrations were (55.27±4.75) ng/ml, (22.31±3.24) ng/ml, (46.23±2.02) ng/ml and (45.92±1.21) ng/ml respectively. Antisense ODN significantly decreased the secretion of FN and ColⅣ in the culture medium (P 0.05). Conclusion Antisense Smad2 ODN can reduce the FN and Col IV secretion in rat mesangial cells cultured with high glucose.