Hydrogen peroxide initiates oxidative stress and proteomic alterations in meningothelial cells

Abstract

Meningothelial cells (MECs) are fundamental cells of the sheaths covering the brain and optic nerve, where they build a brain/optic nerve-cerebral spinal fluid (CSF) barrier that prevents the free flow of CSF from the subarachnoid space, but their exact roles and underlying mechanisms remain unclear. Our attempt here was to investigate the influence elicited by hydrogen peroxide (H2O2) on functional changes of MECs. Our study showed that cell viability of MECs was inhibited after cells were exposed to oxidative agents. Cells subjected to H2O2 at the concentration of 150 µM for 24 h and 48 h exhibited an elevation of reactive oxygen species (ROS) activity, decrease of total antioxidant capacity (T-AOC) level and reduced mitochondrial membrane potential (ΔΨm) compared with control cells. 95 protein spots with more than twofold difference were detected in two dimensional electrophoresis (2DE) gels through proteomics assay following H2O2 exposure for 48 h, 10 proteins were identified through TOF/MS analysis. Among the proteomic changes explored, 8 proteins related to energy metabolism, mitochondrial function, structural regulation, and cell cycle control were downregulated. Our study provides key insights that enhance our understanding of the role of MECs in the pathology of brain and optic nerve disorders.

The role of GAPDH in the selective toxicity of CNP in melanoma cells

“…Recently, it has been published that hydrogen peroxide is both involved in proteomic alterations and in redox signaling [ 23 , 24 ], with oxidation of mostly cysteine residues resulting in functional alterations of the affected proteins [ 25 ]. As we proposed that incubation with CNP led to an increase in hydrogen peroxide, we wanted to test if CNP treatment alters thiol oxidation as H 2 O 2 results in generation of sulfenic (R-SOH), sulfinic (R-SO 2 H) and sulfonic acids (R-SO 3 H).…”

Translatability of In Vitro Stress for Predicting Deamidation and Oxidation Biotransformation on Biotherapeutics

“…19,20 Similarly, hydrogen peroxide is a physiologically relevant agent of oxidative stress, and its addition has been also used to generate oxidative stress in cells. 21,22 However, there have been limited data sets to confirm the in vitro stress translatability to in vivo oxidation specifically for protein therapeutics. 18 In our previous work, we discussed the importance of minimizing artifacts from sample preparation for detecting biotransformation for deamidation.…”

“…A good predictive tool in vitro can potentially allow a gating strategy to trigger biotransformation analyses in vivo . While there have been reports suggesting a good correlation between in vitro stress and in vivo systematic deamidation and isomerization, these reports have limited data sets to provide a comprehensive understanding of translatability. , There have been previous reports of using AAPH stress to model cellular and in vivo oxidative stress. , Similarly, hydrogen peroxide is a physiologically relevant agent of oxidative stress, and its addition has been also used to generate oxidative stress in cells. , However, there have been limited data sets to confirm the in vitro stress translatability to in vivo oxidation specifically for protein therapeutics …”

Synthesis and photochemical properties of caged peroxides for photocontrol of cellular oxidative stress

“…S10, ESI†). To induce MMP depolarisation, cells usually need to be treated with 100–500 μM H 2 O 2 or TBHP, 7,9,29,30 but MitoTBHP could induce significant changes in mitochondrial membrane potential upon photo-irradiation at only 5 μM (Fig. S11, ESI†).…”

“…S10, ESI †). To induce MMP depolarisation, cells usually need to be treated with 100-500 mM H 2 O 2 or TBHP, 7,9,29,30 but MitoTBHP could induce significant changes In conclusion, we designed and synthesised a new caged hydroperoxide, BhcTBHP, consisting of a Bhc phototrigger and TBHP as an effector, and successfully developed membranepermeable AcBhcTBHP and mitochondria-targeted MitoTBHP. These caged compounds rapidly and efficiently released TBHP upon blue light irradiation.…”