AQP4 KO exacerbating renal dysfunction is mediated by endoplasmic reticulum stress and p66Shc and is attenuated by apocynin and endothelin antagonist CPU0213

• Published in: European journal of pharmacology Vol. 721; no. 1-3; pp. 249 - 258
• Main Authors: Cheng, Yu-Si, Dai, De-Zai, Dai, Yin
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 05.12.2013
• Subjects: Acetophenones - antagonists & inhibitors; Animals; AQP4; Aquaporin 4 - deficiency; Aquaporin 4 - genetics; Connexin 43 - genetics; Creatinine - blood; CX43; Down-Regulation - drug effects; eIF-2 Kinase - genetics; Endoplasmic Reticulum Stress - drug effects; Endothelins - antagonists & inhibitors; ETA; Female; Gene Knockout Techniques; Kidney - pathology; Kidney - physiopathology; Male; Malondialdehyde - blood; Matrix Metalloproteinase 9 - genetics; Mice; MMP9; p66Shc; PERK; Pyrazoles - pharmacology; Shc Signaling Adaptor Proteins - metabolism; Src Homology 2 Domain-Containing, Transforming Protein 1; AQP4; ETA; PERK; CX43; MMP9; p66Shc; ET(A)
• ISSN: 0014-2999; 1879-0712
• DOI: 10.1016/j.ejphar.2013.09.028

Aquaporin 4 (AQP4) is essential in normal kidney. We hypothesized that AQP4 knockout (KO) may exacerbate pro-inflammatory factors in the stress induced renal insufficiency. Mechanisms underlying are likely due to activating renal oxidative stress adaptor p66Shc and endoplasmic reticulum (ER) stress that could be mediated by endothelin (ET)—NADPH oxidase (NOX) pathway. AQP4 KO and wild type (WT) mice were randomly divided into 4 groups: control, isoproterenol (1mg/kg, s.c., 5d), and interventions in the last 3 days with either apocynin (NADPH oxidase inhibitor, 100mg/kg, p.o.) or CPU0213 (a dual endothelin receptor antagonist 200mg/kg, p.o.). In addition, HK2 cells were cultured in 4 groups: control, isoproterenol (10−6M), intervened with apocynin (10−6M) or CPU0213 (10−6M). In AQP4 KO mice elevated creatinine levels were further increased by isoproterenol compared to AQP4 KO alone. In RT-PCR, western blot and immunohistochemical assay p66Shc and PERK were significantly increased in the kidney of AQP4 KO mice, associated with pro-inflammatory factors CX40, CX43, MMP-9 and ETA compared to the WT mice. Expression of AQP4 was escalated in isoproterenol incubated HK2 cells, and the enhanced protein of PERK and p-PERK/PERK, and p66shc in vivo and in vitro were significantly attenuated by either apocynin or CPU0213. In conclusion, AQP4 KO deteriorates renal dysfunction due to exacerbating ER stress and p66Shc in the kidney. Either endothelin antagonism or NADPH oxidase blockade partly relieves renal dysfunction through suppressing abnormal biomarkers by APQ4 KO and isoproterenol in the kidney.