TY  - JOUR
AU  - Khan, Alina
AU  - Wan, Tengfei
AU  - Hunter, Catherine
AU  - Schweizer, Pavel
AU  - ishaq, Abbas
AU  - Nagarathanam, Shobha
AU  - Morris, Julia
AU  - Chan, Minyu
AU  - Saretzki, Gabriele
PY  - 2019
DA  - 2019/08/01
TI  - Characterisation of Oxidative Stress, DNA Damage and Inflammation in a Cellular Model of Parkinson’s Disease
JO  - OBM Neurobiology
SP  - 036
VL  - 03
IS  - 03
AB  - Background: Parkinson’s disease (PD) is the second most commonly occurring neurodegenerative disease and is classed as a synucleinopathy due to the critical role of α-synuclein (α-Syn) in its pathology. α-Syn is able to translocate from the cytoplasm to the nucleus and cause DNA damage. Methods: SH-SY5Y cells were stably transfected with plasmids containing wildtype α-Syn and A53T mutant α-Syn as fusion proteins with EGFP and an EGFP only control vector. The cells were treated with hydrogen peroxide (H2O2) and analysed in a differentiated state for the effects of oxidative stress using flow cytometry, DNA damage using H2A.X staining and inflammatory and senescence markers using qPCR.
Results: Cells expressing the A53T mutation exhibited a higher sensitivity to treatment with H2O2 with significantly higher amounts of DNA damage. Importantly, WT α-Syn seemed to have a protective effect in differentiated cells under increased stress with no increase in DNA damage after H2O2 treatment. In line with these results, inflammatory markers of COX-2, IL-6 and TNF-α as well as stress-and senescence markers p21 and p16 were all significantly increased in differentiated treated cells expressing mutated α-Syn while there were no changes in cells expressing wildtype α-Syn. Interestingly, both forms of α-Syn increased the levels of mitochondrial ROS already in untreated conditions while there were no differences between the two α-Syn forms.
Conclusions: Our data demonstrate a higher stress sensitivity of SH-SY5Y neuroblastoma cells harbouring an A53T mutant α-Syn for DNA damage and selected inflammatory marker. In contrast, wildtype α-Syn seemed to provide protection against oxidative stress in a neurone-like cellular model. Thus, these neuroblastoma cells differentiated with retinoid acid seem to be a good model for studying various pathological effects of α-Syn.
SN  - 2573-4407
UR  - https://doi.org/10.21926/obm.neurobiol.1903036
DO  - 10.21926/obm.neurobiol.1903036
ID  - Khan2019
ER  -