Proteomics and Toxicity Analysis of Spinal-Cord Primary Cultures upon Hydrogen Sulfide Treatment.

Viviana Greco,Victor Corasolla Carregari,Alida Spalloni,Patrizia Longone,Andrea Urbani,Luisa Pieroni,Silvia Persichilli,Nicola Mercuri

doi:10.3390/antiox7070087

Abstract

Hydrogen sulfide (H2S) is an endogenous gasotransmitter recognized as an essential body product with a dual, biphasic action. It can function as an antioxidant and a cytoprotective, but also as a poison with a high probability of causing brain damage when present at noxious levels. In a previous study, we measured toxic liquoral levels of H2S in sporadic amyotrophic lateral sclerosis (ALS) patients and in the familial ALS (fALS) mouse model, SOD1G93A. In addition, we experimentally demonstrated that H2S is extremely and selectively toxic to motor neurons, and that it is released by glial cells and increases Ca2+ concentration in motor neurons due to a lack of ATP. The presented study further examines the effect of toxic concentrations of H2S on embryonic mouse spinal-cord cultures. We performed a proteomic analysis that revealed a significant H2S-mediated activation of pathways related to oxidative stress and cell death, particularly the Nrf-2-mediated oxidative stress response and peroxiredoxins. Furthermore, we report that Na2S (a stable precursor of H2S) toxicity is, at least in part, reverted by the Bax inhibitor V5 and by necrostatin, a potent necroptosis inhibitor.

Highlights

In 2015 [1], we showed that hydrogen sulfide (H2 S) reaches toxic levels in the cerebrospinal fluid (CSF) of sporadic amyotrophic lateral sclerosis (ALS) patients
To understand the effects of H2 S-mediated toxicity on neuronal cultures, we performed a deep proteomic investigation in order to obtain the expression profile of proteins whose levels change in response to treatment with Na2 S
To investigate the different protein profiles with and without Na2 S, we used protein extracts from spinal-cord cultures not treated and treated with Na2 S at a concentration of 200 μM, in which the cytotoxicity of motor neurons was markedly enhanced, as we previously demonstrated in Davoli et al [1]

Summary

Introduction

In 2015 [1], we showed that hydrogen sulfide (H2 S) reaches toxic levels in the cerebrospinal fluid (CSF) of sporadic amyotrophic lateral sclerosis (ALS) patients. Like carbon monoxide and nitric oxide, is recognized as a cellular gaseous mediator and a neurotransmitter [2,3,4]. It is produced via cysteine catabolism by the cytoplasmic enzymes, cystathionine-β-synthase (CBS) and cystathionine-γ-lyase (CSE), and via 3-mercaptopyruvate catabolism by 3-mercaptopyruvate sulfurtransferase (3-MST) [5]. On one hand, H2 S showed protective abilities in cellular and animal models of Parkinson’s disease (PD) and Alzheimer’s disease (AD).

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Conclusion