EXPRESSION OF TRANSFORMING GROWTH-FACTOR-BETA-1 DURING DIABETIC RENAL HYPERTROPHY

Expression of transforming growth factor-beta 1 during diabetic renal hypertrophy. Experimental type I diabetes mellitus is characterized by an early increase in kidney weight and glomerular volume, but changes in gene expression accompanying diabetic renal growth have not been fully elucidated. In the current study, total RNA was extracted from renal cortex and isolated glomeruli of streptozotocin-induced diabetic rats 24 hours, 48 hours, 96 hours, one and two weeks after the onset of hyperglycemia (blood glucose >15 mmol/liter), insulin-treated diabetic rats (blood glucose <6.0 mmol/liter), and normal rats. RNA samples were reverse transcribed (RT) and subjected to polymerase chain reaction (PCR) amplication with specific 5' and 3' primers for rat transforming growth factor (TGF-beta 1) and beta-actin. RT-PCR analysis revealed that glomerular TGF-beta 1 mRNA levels increased relative to beta-actin as early as 24 hours after the onset of hyperglycemia, reaching a plateau after 96 hours that was sustained at one and two weeks. In cortical samples, TGF-beta 1 mRNA levels increased less abruptly, reaching a peak one week after the onset of hyperglycemia. Intensive insulin treatment to normalize blood glucose levels attenuated the rise in glomerular and renal cortical TGF-beta 1 mRNA. Cryostat sections of rat kidneys were immunostained for TGF-beta 1 utilizing a polyclonal anti-porcine TGF-beta 1 antibody and semiquantitative scoring of TGF-beta 1 immunostaining revealed a twofold increase in diabetic glomeruli after two weeks compared to normal glomeruli. Increased segmental immunostaining for TGF-beta 1 was also evident in cortical tubules of diabetic rats. These studies establish that TGF-beta 1 expression in the kidney increases during the phase of rapid renal hypertrophy in diabetic rats. Normalization of blood glucose levels with insulin treatment attenuates the increase in TGF-beta 1 expression.