H₂S contributed from CSE during cellular senescence suppresses inflammation and nitrosative stress

Aging involves the time-dependent deterioration of physiological functions attributed to various intracellular and extracellular factors. Cellular senescence is akin to aging and involves alteration in redox homeostasis. This is primarily marked by increased reactive oxygen/nitrogen species (ROS/RNS), inflammatory gene expression, and senescence-associated beta-galactosidase activity, all hallmarks of aging. It is proposed that gasotransmitters which include hydrogen sulfide (H₂S), carbon monoxide (CO), and nitric oxide (NO), may affect redox homeostasis during senescence. H₂S has been independently shown to induce DNA damage and suppress oxidative stress. While an increase in NO levels during aging is well established, the role of H₂S has remained controversial. To understand the role of H₂S during aging, we evaluated H₂S homeostasis in non-senescent and senescent cells, using a combination of direct measurements with a fluorescent reporter dye (WSP-5) and protein sulfhydration analysis. The free intracellular H₂S and total protein sulfhydration levels are high during senescence, concomitant to cystathionine gamma-lyase (CSE) expression induction. Using lentiviral shRNA-mediated expression knockdown, we identified that H₂S contributed by CSE alters global gene expression, which regulates key inflammatory processes during cellular senescence. We propose that H₂S decreases inflammation during cellular senescence by reducing phosphorylation of IκBα and the p65 subunit of nuclear factor kappa B (NF-κB). H₂S was also found to reduce NO levels, a significant source of nitrosative stress during cellular senescence. Overall, we establish H₂S as a key gasotransmitter molecule that regulates inflammatory phenotype and nitrosative stress during cellular senescence.