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Switch from ACE to Chymase mRNA Expression in Diabetes in Enriched Renal Vascular Tissues Harvested by Manual Sieving

Our previous work supports a major role for ACE‐independent intrarenal ANGII formation on microvascular function in type II diabetes (Park et al. AJP 298, 2010). The current studies tested the hypothesis that a switch from ACE to chymase gene expression occurs in the diabetic renal vasculature. Kidneys were removed from anesthetized diabetic (db/db; n=7) and control (db/m; n=15) mice and placed in a 100 micron sieve. Using a spatula, slow, grating motions were applied over the sieve to remove the tubules, while preserving the vessels (Schneider et al., AJP 299, 2010). Protein expression of α‐smooth muscle actin (ASMA) and ACE was measured by Western blot. Chymase and ACE mRNA expression was measured by RT‐PCR. Protein expression of ASMA/β‐actin was increased 14.2 ± 0.5 fold in renal vessels (n=4) relative to renal cortex (n=4). In contrast, expression of ACE/β‐actin was increased 16.5 ± 0.7 fold in renal cortex compared to renal vessels. Further, chymase/β‐actin mRNA expression was elevated 5.1 ± 1.4 fold and ACE/β‐actin mRNA was decreased 0.42 ± 0.08 fold in diabetic compared to control vessels. Sieving results in renal vessels with augmented ASMA and nominal ACE protein expression, confirming vascular enrichment with minimal tubular contamination. In conclusion, enhanced chymase and decreased ACE mRNA expression in diabetic vessels supports a molecular switch that may contribute to renal disease progression.