Zoltán Kiss 1,2, Beáta Szebeni 2, Domonkos Pap 2, István M. Takács 1, Apor Veres-Székely 1, Marianna Gyurászová 1,3, Erna Sziksz 2, Leonóra Balicza-Himer 2, Attila Szabó 1,2, Ádám Vannay 1,2
1 1st Department of Pediatrics, Semmelweis University
2 MTA-SE Pediatric and Nephrology Research Group
3 Institute of Molecular Biomedicine, Faculty of Medicine, Comenius University, Bratislava, Slovakia
Background: Ischaemia-reperfusion (I/R) induced acute renal failure (ARF) has a high risk of morbidity and mortality, and remains a major medical problem without reliable pharmacological intervention. Our aim was to analyse the underling proteomic changes related to I/R.
Methods: Two-dimensional differential gel electrophoresis mass spectrometry analysis were carried out on kidney tissue samples from an I/R model of male Wistar rats and respective sham controls. Kidney epithelial cells (HEK-293) were transfected with a plasmid constructed to constitutively express the Parkinson disease 7 (PARK7) protein. H2O2 treatment were used in the in vitro experiments on the transfected and respective control cells.. Cell survival was measured by the LDH method, and gene expression was measured by RT-PCR method.
Results: Bioinformatic analysis of the proteomic results revealed PARK7 as a central factor related to I/R induced oxidative stress of the kidney. Our experiments on HEK-239 cells overexpressing PARK7 showed, that elevated level of PARK7 improved the survival of the cells following H2O2 treatment compared to the respective controls. This phenomenon was confirmed by decreased expression of the pro-apoptotic Bcl-2-associated X protein (BAX) in the H2O2 treated cells overexpressing PARK7 compared to the controls. Furthermore, the expression of antioxidant enzyme NAD(P)H dehydrogenase (NQO1) increased in the H2O2 treated cells overexpressing PARK7 compared to the controls.
Conclusions: Overexpression of PARK7 attenuated the H2O2 induced cell death presumably through the altered expression of NQO1. Our data suggest, that besides the well-known intrinsic chaperon functions, PARK7 affect the survival or renal tubular epithelial cells by activating antioxidant signalling pathways.
Fundings: This project was supported by the ÚNKP-18-3-III-SE-29 and the ÚNKP-18-4-SE-109. New National Excellence Program of the Ministry of Human Capacities, the János Bolyai Research Scholarship of the Hungarian Academy of Sciences, the grants OTKA K116928., 20382-3/2018 FEKUTSTRAT, and the Semmelweis Science Innovation Fund.
Károly Rácz School of PhD Studies
Clinical Medicine PhD School
03. Prevention of Chronic Diseases in Childhood
Supervisor: Dr.Gábor Veres
UNKP supervisor: Dr. Ádám Vannay