Downregulation of endothelin A receptor (ETaR) ameliorates renal ischemia reperfusion injury by increasing nitric oxide production

AIMS:To investigate the protective effects of downregulating ETaR expression on renal ischemia reperfusion injury (IRI). MAIN METHODS:The renal IRI model was generated by clamping the left renal artery for 60 min followed by nephrectomy of the right kidney. ETaR siRNA were perfused through the renal artery during ischemia. HE staining was performed to assess histological injury. PCR was performed to determine the expression of NF-κb, TNF-α, IFN-γ, IL-6 and TGF-β. ELISA was used to determine the levels of ET-1, TGF-β and eNOS. The level of nitric oxide (NO) was tested by the NO detection kit. The expression of PI3K, Akt, sGC and PKG were evaluated by western blot. KEY FINDINGS:ETaR siRNA treatment reduced the levels of serum creatinine and urea nitrogen, decreased the number of apoptotic cells, and ameliorated histological damage after IRI. PCR results demonstrated that IRI increased mRNA levels of inflammatory factors, which were inhibited by ETaR siRNA treatment. ELISA result showed that ETaR siRNA decreased the levels of ET-1, TGF-β and eNOS in the renal tissues after IRI. Western blot results demonstrated that ETaR siRNA activated the PI3K/Akt and sGC/PKG signaling pathway. Conversely, the NOS inhibitor, L-NAME, reversed the effects of ETaR siRNA treatment. SIGNIFICANCE:ETaR siRNA treatment inhibited inflammatory response and improved renal function after renal IRI. The underlying mechanisms of ETaR siRNA treatment may be through increasing eNOS activity through PI3K/Akt signaling, which subsequently increased NO production. The increased NO reduces the expression of ET-1 by inhibiting transcription of ET-1-associated genes via the sGC/PKG signaling pathway.