Hydrogen peroxide modifies both activity and isoforms of acetylcholinesterase in human neuroblastoma SH-SY5Y cells

Abstract

The involvement of cholinergic system and the reactive oxygen species (ROS) in the pathogenesis of some degenerative diseases has been widely reported; however, the specific impact of hydrogen peroxide (H2O2) on the acetylcholinesterase (AChE) activity as well as AChE isoform levels has not been clearly established. Hence, the purpose of present study is to clarify whether H2O2 alters these parameters.Human neuroblastoma SH-SY5Y cells were treated with H2O2 (1–1000 µM) for 24 h and AChE activity and AChE and cytochrome c levels were evaluated. AChE activity was strongly increased from 1 µM to 1000 µM of H2O2. The results of the kinetic study showed that H2O2 affected Vmax but not Km; and also that H2O2 changed the sigmoid kinetic observed in control samples to hyperbolic kinetic. Thus, results suggest that H2O2 acts as an allosteric activators. In addition, H2O2, (100–1000 µM) reduced the total AChE content and modified its isoform profile (mainly 50-, 70-, and 132-kDa)·H2O2 from 100 µM to 1000 µM induced cytochrome c release confirming cell death by apoptosis. All these results together suggest: a) the involvement of oxidative stress in the imbalance of AChE; and b) treatment with antioxidant agents may be a suitable strategy to protect cholinergic system alterations promoted by oxidative stress.

Hydrogen peroxide-induced oxidative stress, acetylcholinesterase inhibition, and mediated brain injury attenuated byThymus algeriensis

“…These findings confirm the studies done by Kindl et al, showing that many Thymus species growing in Croatia exhibited anti-AChE activity in a dose dependent manner. This data coincides with previous observations that administration of H 2 O 2 to human neuroblastoma SH-SY5Y cell line modifies the isoform profile of AChE and its activity, which was strongly increased from 1 µM to 1000 µM of H 2 O 2 (Garcimartín et al, 2017).…”

“…Acetylcholinesterase (AChE), as a key enzyme in the cholinergic nervous system (Mathew and Subramanian, 2014), hydrolyzes acethylcholine (ACh) to choline and acetate, thereby terminating the effect of this neurotransmitter at cholinergic synapses (Vladimir-Knežević et al, 2014). Treatment of human neuroblastoma cells with H 2 O 2 modified AChE isoform profile and reduced its available amount and/ or cellular death by apoptosis (Garcimartín et al, 2017). Numerous phytoconstituents and promising plant species have been reported as…”

Micro- and nano-plastics activation of oxidative and inflammatory adverse outcome pathways

“…Simultaneous detection of MP/NPs induced oxidative stress and AChE inhibition further supports such possibility [ 11 , 48 ]. Direct relationship between H 2 O 2 and modifications of AChE metabolism and activity was established recently, when it was determined that H 2 O 2 decreased AChE levels by allosteric effector action on AChE structure and changes of isoform profile in SH-SY5Y cells confirming that increase in at least one ROS could promote disturbances in cholinergic system of neural cells [ 67 ].…”

Synthesis, characterization and pharmacological evaluation of quinoline derivatives and their complexes with copper(ΙΙ) in in vitro cell models of Alzheimer's disease

“…Acetylcholinesterase, the main target of current AD drugs, is also involved indirectly in the Αβ and oxidative stress hypotheses of AD. For example, Aβ peptide aggregation is promoted by AChE via its peripheral anionic site (PAS) [52][53][54], while, oxidative stress may alter the levels and/or activity of AChE directly or indirectly [55,56]. Interestingly, treatment of SH-SY5Y cells with H2O2 results in allosteric activation of AChE characterized by an increase in Vmax with no changes in Km and, importantly, lack of dose-dependent effects of H2O2 in culture [55].…”