Hydrogen sulfide as an anti-calcification stratagem in human aortic valve: Altered biogenesis and mitochondrial metabolism of H2S lead to H2S deficiency in calcific aortic valve disease

Abstract

Hydrogen sulfide (H₂S) was previously revealed to inhibit osteoblastic differentiation of valvular interstitial cells (VICs), a pathological feature in calcific aortic valve disease (CAVD). This study aimed to explore the metabolic control of H₂S levels in human aortic valves. Lower levels of bioavailable H₂S and higher levels of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) were detected in aortic valves of CAVD patients compared to healthy individuals, accompanied by higher expression of cystathionine γ-lyase (CSE) and same expression of cystathionine β-synthase (CBS). Increased biogenesis of H₂S by CSE was found in the aortic valves of CAVD patients which is supported by increased production of lanthionine. In accordance, healthy human aortic VICs mimic human pathology under calcifying conditions, as elevated CSE expression is associated with low levels of H₂S. The expression of mitochondrial enzymes involved in H₂S catabolism including sulfide quinone oxidoreductase (SQR), the key enzyme in mitochondrial H₂S oxidation, persulfide dioxygenase (ETHE1), sulfite oxidase (SO) and thiosulfate sulfurtransferase (TST) were up-regulated in calcific aortic valve tissues, and a similar expression pattern was observed in response to high phosphate levels in VICs. AP39, a mitochondria-targeting H₂S donor, rescued VICs from an osteoblastic phenotype switch and reduced the expression of IL-1β and TNF-α in VICs. Both pro-inflammatory cytokines aggravated calcification and osteoblastic differentiation of VICs derived from the calcific aortic valves. In contrast, IL-1β and TNF-α provided an early and transient inhibition of VICs calcification and osteoblastic differentiation in healthy cells and that effect was lost as H₂S levels decreased. The benefit was mediated via CSE induction and H₂S generation. We conclude that decreased levels of bioavailable H₂S in human calcific aortic valves result from an increased H₂S metabolism that facilitates the development of CAVD. CSE/H₂S represent a pathway that reverses the action of calcifying stimuli.

Keywords: Arteriosclerosis; Chronic kidney disease; Hydrogen sulfide; Mitochondrial H(2)S catabolism; Phosphate; Valvular inflammation; Vascular calcification.