Uremic Toxin Indoxyl Sulfate Impairs Hydrogen Sulfide Formation in Renal Tubular Cells.

Hydrogen sulfide (H 2 S) was the third gasotransmitter to be recognized as a cytoprotectant. A recent study demonstrated that exogenous supplementation of H 2 S ameliorates functional insufficiency in chronic kidney disease (CKD). However, how the H 2 S system is impaired by CKD has not been elucidated. The uremic toxin indoxyl sulfate (IS) is known to accumulate in CKD patients and harm the renal tubular cells. This study therefore treated the proximal tubular cells, LLC-PK 1 , with IS to see how IS affects H 2 S formation. Our results showed that H 2 S release from LLC-PK 1 cells was markedly attenuated by IS when compared with control cells. The H 2 S donors NaHS and GYY-4137 significantly attenuated IS-induced tubular damage, indicating that IS impairs H 2 S formation. Interestingly, IS downregulated the H 2 S-producing enzymes cystathionine β-synthase (CBS), cystathionine γ-lyase (CSE), and 3-mercaptopyruvate sulfurtransferase (3-MST), and these effects could be reversed by inhibition of the IS receptor, aryl hydrocarbon receptor (AhR). As transcription factor specificity protein 1 (Sp1) regulates the gene expression of H 2 S-producing enzymes, we further showed that IS significantly decreased the DNA binding activity of Sp1 but not its protein expression. Blockade of AhR reversed low Sp1 activity caused by IS. Moreover, exogenous H 2 S supplementation attenuated IS-mediated superoxide formation and depletion of the cellular glutathione content. These results clearly indicate that IS activates AhR, which then attenuates Sp1 function through the regulation of H 2 S-producing enzyme expression. The attenuation of H 2 S formation contributes to the low antioxidant defense of glutathione in uremic toxin-mediated oxidative stress, causing tubular cell damage.