Clinical and basic studies have revealed that the renin-angiotensin system (RAS) plays crucial roles in the development of kidney injury during progression of type 1 diabetes mellitus (T1DM) whereas the activity of systemic RAS is suppressed in T1DM. Thus, intrarenal RAS is increasingly being recognized as an important factor in the development of diabetic nephropathy. We recently demonstrated expression of TRPA1, an oxidative stress-sensitive calcium channel, in renal proximal tubular cells (PTC) and TRPA1 mediates angiotensinogen (AGT) augmentation in the cells. However, regulation and function of the intrarenal TRPA1 in T1DM have not been elucidated. Therefore, this study was performed to evaluate intrarenal TRPA1 expression and its contribution to AGT augmentation under high glucose condition. Streptozotocin (STZ, 200 mg/kg) was intraperitoneally injected to male C57BL/6 mice. Intrarenal TRPA1 expression, kidney AGT, urinary AGT, urinary 8-isoprostane, a maker of oxidative stress, and blood glucose levels were determined one week after STZ injection. Furthermore, contribution of TRPA1 to AGT regulation was tested by using mouse PTC which was cultured with normal (1.0 g/L) or high (4.5 g/L) glucose media. Under these conditions, PTC was treated with 100 μM hydrogen peroxide. Blood glucose and urinary 8-isoprostane levels were higher in the STZ-treated mice compared with control mice. TRPA1 mRNA levels were augmented in the kidney of STZ-treated mice (2.26 ± 0.2, ratio to control mice) accompanied by increases in kidney AGT and urinary AGT levels. TPRA1 upregulation was also observed in PTC which was cultured with the high glucose medium (1.55 ± 0.11, ratio to the normal glucose). Hydrogen peroxide stimulated AGT expression in PTC. The AGT elevation was enhanced in the cultured cells with the high glucose (1.52 ± 0.13-fold in the normal glucose medium vs. 1.97 ± 0.29-fold in the high glucose medium). Treatment with a TRPA1 antagonist attenuated the enhanced AGT augmentation in PTC. These results demonstrate that intrarenal TRPA1 mediates enhanced AGT upregulation under high glucose condition and suggest that intrarenal TRPA1 is implicated in intrarenal RAS activation leading to the development of T1DM-associated diabetic nephropathy.
Abnormal glutathione metabolism and increased cytotoxicity caused by H 2 O 2 in human umbilical vein endothelial cells cultured in high glucose medium