Alteration in renin‐angiotensin system (RAS) has been implicated in the pathophysiology of diabetic kidney disease (DKD). The deleterious actions of angiotensin II (Ang II) could be antagonized by the formation of Ang‐(1–7), generated by the actions of angiotensin‐converting enzyme 2 (ACE2) and neprilysin (NEP). NEP degrades several peptides, including natriuretic peptides, bradykinin, amyloid beta, and Ang I. Although combination of Ang II receptor and NEP inhibitor treatment benefits patients with heart failure, the role of NEP in renal pathophysiology is a matter of active research. NEP pathway is a potent enzyme in Ang I to Ang‐(1–7) conversion in the kidney of ACE2‐deficient mice, suggesting a renoprotective role of NEP. The aim of the study is to test the hypothesis that chronic hyperglycemia downregulates renal NEP protein expression and activity in db/db diabetic mice and treatment with rosiglitazone normalizes hyperglycemia, renal NEP expression, and attenuates albuminuria. Mice received rosiglitazone (20 mg kg−1 day−1) for 10 weeks. Western blot analysis, immunohistochemistry, and enzyme activity revealed a significant decrease in renal and urinary NEP expression and activity in 16‐wk db/db mice compared with lean control (p < .0001). Rosiglitazone also attenuated albuminuria and increased renal and urinary NEP expressions (p < .0001). In conclusion, data support the hypothesis that diabetes decreases intrarenal NEP, which could have a pivotal role in the pathogenesis of DKD. Urinary NEP may be used as an index of intrarenal NEP status. The renoprotective effects of rosiglitazone could be mediated by upregulation of renal NEP expression and activity in db/db diabetic mice.

Activation of the renin angiotensin system plays a pivotal role in the regulation of blood pressure, which is mainly attributed to the formation of angiotensin-II (Ang II). The actions of Ang II are mediated through binding to the Ang-II type 1 receptor (AT1R) which leads to increased blood pressure, fluid retention, and aldosterone secretion. In addition, Ang II is also involved in cell injury, vascular remodeling, and inflammation. The actions of Ang II could be antagonized by its conversion to the vasodilator peptide Ang (1–7), partly generated by the action of angiotensin converting enzyme 2 (ACE2) and/or neprilysin (NEP). Previous studies demonstrated increased urinary ACE2 shedding in the db/db mouse model of diabetic kidney disease. The aim of the study was to investigate whether renal and urinary ACE2 and NEP are altered in the 2K1C Goldblatt hypertensive mice. Since AT1R is highly expressed in the kidney, we also researched the effect of global deletion of AT1R on renal and urinary ACE2, NEP, and kidney injury marker (KIM-1). Hypertension and albuminuria were induced in AT1R knock out (AT1RKO) and WT mice by unilateral constriction of the renal artery of one kidney. The 24 h mean arterial blood pressure (MAP) was measured using radio-telemetry. Two weeks after 2K1C surgery, MAP and albuminuria were significantly increased in WT mice compared to AT1RKO mice. Results demonstrated a correlation between MAP and albuminuria. Unlike db/db diabetic mice, ACE2 and NEP expression and activities were significantly decreased in the clipped kidney of WT and AT1RKO compared with the contralateral kidney and sham control (p &lt; 0.05). There was no detectable urinary ACE2 and NEP expression and activity in 2K1C mice. KIM-1 was significantly increased in the clipped kidney of WT and AT1KO (p &lt; 0.05). Deletion of AT1R has no effect on the increased urinary KIM-1 excretion detected in 2K1C mice. In conclusion, renal injury in 2K1C Goldblatt mouse model is associated with loss of renal ACE2 and NEP expression and activity. Urinary KIM-1 could serve as an early indicator of acute kidney injury. Deletion of AT1R attenuates albuminuria and hypertension without affecting renal ACE2, NEP, and KIM-1 expression.

Alteration in renin-angiotensin system has been implicated in the pathophysiology of diabetic kidney disease. The deleterious actions of angiotensin II (Ang II) could be antagonized by the formation of Ang (1-7) partly generated by the actions of angiotensin converting enzyme 2 (ACE2) and neprilysin (NEP). NEP is a member of the zinc-containing metallopeptidase family, and has a main role in the degradation of several peptides, including natriuretic peptides, bradykinin, amyloid beta, and Ang I. Our previous studies demonstrated increased shedding of renal ACE2 in db/db mouse model of type 2 diabetes. We also showed that treatment with the PPAR_ agonist, rosiglitazone normalized hyperglycemia, decreased urinary ACE2 and attenuated albuminuria in db/db mice. The aim of the study was to test the hypothesis that hyperglycemia down-regulates renal NEP in db/db diabetic mice and that treatment with rosiglitazone will normalize renal NEP expression and activity. Seven-week-old db/db male mice were subjected to rosiglitazone treatment (20 mg/kg/day) for 10 weeks. Treatment with rosiglitazone significantly lowered blood glucose levels in db/db mice (p<0.001). Western blot analysis and immunohistochemistry demonstrated a significant decrease in renal and urinary NEP protein expression in 17wk db/db mice compared to lean control mice (p<0.0001). Treatment of db/db mice with rosiglitazone attenuated albuminuria, increased renal NEP protein expression and activity. In conclusion, the renoprotective effects of rosiglitazone could be mediated by up-regulation of renal NEP expression and activity in db/db mice. Alteration in the balance between Ang II and Ang (1-7) forming enzymes could contribute to the development of albuminuria in db/db diabetic mice.