The epidemiological features of hydrogen sulfide (H2S) intoxication have changed over the last few years. While H2S still poses a persistent risk as a chemical hazard for workers in gas or oil industry, H2S poisoning has become a new method of suicide with an incidence which has increased at an alarming rate in many countries including the US. H2S toxicity results from the combination of a direct inhibition of the mitochondrial activity of neurons and the consequences of the cardio-respiratory failure (apnea and shock) on oxygen delivery. H2S is lethal within minutes at a concentration of less than 0.1 %;at lower concentrations, H2S produces an acute coma leading to memory and cognitive deficits in the surviving patients. Our recent findings of the very fast disappearance of free/soluble H2S following hydrogen sulfide exposure represents a major limitation for using, after the intoxication, metallo-compounds, i.e. the only family of antidotes with a proven efficacy during H2S exposure. Indeed, the benefit of Fe3+or Co2+ compounds currently proposed against H2S toxicity relies on their ability to trap free/soluble H2S in the blood, as for instance with methemoglobin, or inside cells, with Hydroxocobalamin (HyCo). However, these agents have a limited action against the pool of H2S combined on proteins and have no effect against the ischemic consequences of H2S-induced circulatory failure. Based on our preliminary data on the effects of Methylene Blue (MB) during H2S poisoning and the remarkable protective effects of MB on post-anoxic or post-toxic brain injury, we are proposing to use MB for the treatment of H2S poisoning. This approach represents a novel paradigm as it specifically counteracts the cellular and mitochondrial dysfunctions induced by H2S. In this project we will determine the ability of MB to prevent the short and long-term effects of H2S- induced brain injury, when administered after an acute H2S intoxication in a rat model of reversible coma. We have developed this model to mimic the most common type of non-lethal form of H2S poisoning in humans. The short-term effects (first 12 hours) of MB following H2S-induced coma will be evaluated by physiological and clinical parameters of post-anoxic injury and brain markers of hypoxia or apoptosis. In a separate group of rats, the long-term effects on memory and cognitive functions will be evaluated in non-anesthetized animals trained to perform tests in a Morris Water Maze after acute or repetitive administrations of MB following an episode of sulfide poisoning-induced coma. MB will be administered alone or combined with HyCo to determine the potential synergy of these 2 antidotes against H2S poisoning. The results of these studies will lead to a novel strategy using an agent counteracting the direct consequences of H2S poisoning. Since MB appears to be also effective against other forms of mitochondrial poisoning, these results may have implications beyond the treatment of H2S intoxication.

Public Health Relevance

Hydrogen sulfide (H2S) poisoning, which remains one of the most dreadful occupational hazards, has become a method of suicide causing an increasing number of new fatal intoxications each year in the US. The window of opportunity for first responders to administer the currently proposed antidotes, which are able to trap H2S, is extremely limited due to the very rapid disappearance of free H2S from the body;so little benefit is to be expected after a few minutes with the traditional treatment. We have recently found that Methylene Blue (MB) infusion can reduce the cardiovascular depression induced by H2S;this proposal will determine, in adult rats, the effects of MB injected after H2S intoxication on the long-term neurological outcome following H2S-induced coma.

National Institute of Health (NIH)
National Institute of Neurological Disorders and Stroke (NINDS)
Exploratory/Developmental Grants (R21)
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Special Emphasis Panel (ZRG1-MDCN-B (55))
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Yeung, David
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Pennsylvania State University
Internal Medicine/Medicine
Schools of Medicine
United States
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Cheung, Joseph Y; Wang, JuFang; Zhang, Xue-Qian et al. (2018) Methylene Blue Counteracts H2S-Induced Cardiac Ion Channel Dysfunction and ATP Reduction. Cardiovasc Toxicol 18:407-419
Judenherc-HaouzI, Annick; Sonobe, Takashi; Bebarta, Vikhyat S et al. (2018) On the Efficacy of Cardio-Pulmonary Resuscitation and Epinephrine Following Cyanide- and H2S Intoxication-Induced Cardiac Asystole. Cardiovasc Toxicol 18:436-449
Imamura, Fumiaki; Cooper, Timothy K; Hasegawa-Ishii, Sanae et al. (2017) Hydrogen Sulfide Specifically Alters NAD(P)H Quinone Dehydrogenase 1 (NQO1) Olfactory Neurons in the Rat. Neuroscience 366:105-112
Sonobe, Takashi; Haouzi, Philippe (2016) Sulfide Intoxication-Induced Circulatory Failure is Mediated by a Depression in Cardiac Contractility. Cardiovasc Toxicol 16:67-78
Sonobe, Takashi; Haouzi, Philippe (2016) H2S concentrations in the heart after acute H2S administration: methodological and physiological considerations. Am J Physiol Heart Circ Physiol 311:H1445-H1458
Judenherc-Haouzi, Annick; Zhang, Xue-Qian; Sonobe, Takashi et al. (2016) Methylene blue counteracts H2S toxicity-induced cardiac depression by restoring L-type Ca channel activity. Am J Physiol Regul Integr Comp Physiol 310:R1030-44
Haouzi, Philippe; Sonobe, Takashi; Judenherc-Haouzi, Annick (2016) Developing effective countermeasures against acute hydrogen sulfide intoxication: challenges and limitations. Ann N Y Acad Sci 1374:29-40
Haouzi, Philippe (2016) Is exogenous hydrogen sulfide a relevant tool to address physiological questions on hydrogen sulfide? Respir Physiol Neurobiol 229:5-10
Sonobe, Takashi; Haouzi, Philippe (2015) H2S induced coma and cardiogenic shock in the rat: Effects of phenothiazinium chromophores. Clin Toxicol (Phila) 53:525-39
Haouzi, Philippe; Sonobe, Takashi (2015) Cardiogenic shock induced reduction in cellular O2 delivery as a hallmark of acute H2S intoxication. Clin Toxicol (Phila) 53:416-7

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