Investigating the Effect of Hydrogen Sulfide (H2S) on Hydrogen Peroxide Treated Hepatocytes

The liver is the largest visceral organ in the human body and plays a major role in multiple metabolic functions, including biotransformation, protein synthesis, and cholesterol metabolism. Hydrogen sulfide (H2S) may play a vital role in mitigating the effects of liver inflammation. Previous studies have shown that a deficiency in two H2S producing enzymes may lead to inflammation in the liver.

Amino acid-based C-substituted N-thiocarboxyanhydrides (NTAs) are H2S releasing-agents. Breakdown of the NTAs results in carbonyl sulfide (COS), which is converted to H2S by carbonic anhydrase, an enzyme that is naturally produced in eukaryotic cells. Different NTAs have various H2S releasing times and the remaining byproduct is the original amino acid from which it was derived.

The goal of this investigation is to determine the effects of H2S on mitigating liver inflammation after administration of hydrogen peroxide (H2O2), a known inflammatory agent. To investigate the effects, we assembled collagen Type I gels and cultured primary rat hepatocytes (primary liver cell type). To induce reactive oxygen species, we administered H2O2 to our cultures at varying concentrations. To enhance inflammation within our cultures we deprived the cells of media prior to H2O2 administration, based on previous reports. After H2O2 administration, 2 NTA solutions (Valine-NTA and Alanine-NTA) were administered to the cultures at two concentrations (100 μM and 250 μM) to investigate how H2S affected the cells. To determine the effects, we collected media every 24 hours for analysis and lysed cells for DNA quantification. Our future work includes investigating how the NTAs affect other hepatic functions after H2O2 administration.