INTRODUCTION: Angiotensin converting enzyme (ACE) 2 acts as a negative regulator of the renin angiotensin system. It has been demonstrated that ACE2 pharmacology inhibition or genetic deletion worsens renal lesions. Our aim was to study the effect of ACE2 deletion on fibrosis as a marker of renal damage in primary proximal tubular cells from NOD mice in a pre-diabetic state.
METHODS: NOD-ACE2-/- and NOD-ACE2+/+ female mice at 12 weeks of age were analyzed. Renal cortex was digested in culture media with collagenase type II. Cells separation was performed through to 100µm- and 80µm-sieves. Obtained cells were cultured in DMEM-F12 media and sub-cultured for 4 times. Cells characterization was performed by SGLT-2 and AQP1 gene expression as positive markers and AQP2 gene expression as a negative marker. Gene expression of collagen-I, collagen-IV, fibronectin, and α-smooth muscle actin (SMA) was determined by qPCR.
RESULTS: As expected, RNA isolated from primary proximal tubular cells wild-type and knockout for ACE2 were positive for SGLT-2 and AQP1 gene expression and negative for AQP2 gene expression. Interestingly, ACE2 knockout cells presented increased collagen-IV and a-SMA gene expression as compared to wild-type cells. CONCLUSIONS: ACE2 deletion in proximal tubular cells may contribute to renal fibrosis. The lack of ACE2 may result in the accumulation of angiotensin II (ANGII) and the stimulation of the ANGII/AT1R axis and subsequent tubular damage.
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