Synthesis of Reactive Sulfur Species in Cultured Vascular Endothelial Cells after Exposure to TGF-β1: Induction of Cystathionine γ-Lyase and Cystathionine β-Synthase Expression Mediated by the ALK5-Smad2/3/4 and ALK5-Smad2/3-ATF4 Pathways

Abstract

Transforming growth factor-β1 (TGF-β1) occurs at high levels at damage sites of vascular endothelial cell layers and regulates the functions of vascular endothelial cells. Reactive sulfur species (RSS), such as cysteine persulfide, glutathione persulfide, and hydrogen persulfide, are cytoprotective factors against electrophiles such as reactive oxygen species and heavy metals. Previously, we reported that sodium trisulfide, a sulfane sulfur donor, promotes vascular endothelial cell proliferation. The objective of the present study was to clarify the regulation and significance of RSS synthesis in vascular endothelial cells after exposure to TGF-β1. Bovine aortic endothelial cells in a culture system were treated with TGF-β1 to assess the expression of intracellular RSS, the effect of RSS on cell proliferation in the presence of TGF-β1, induction of RSS-producing enzymes by TGF-β1, and intracellular signal pathways that mediate this induction. The results suggest that TGF-β1 increased intracellular RSS levels to modulate its inhibitory effect on proliferation. The increased production of RSS, probably high-molecular-mass RSS, was due to the induction of cystathionine γ-lyase and cystathionine β-synthase, which are RSS-producing enzymes, and the induction was mediated by the ALK5-Smad2/3/4 and ALK5-Smad2/3-ATF4 pathways in vascular endothelial cells. TGF-β1 regulates vascular endothelial cell functions such as proliferation and fibrinolytic activity; intracellular high-molecular-mass RSS, which are increased by TGF-β1, may modulate the regulation activity in vascular endothelial cells.

The Dipeptide Gly-Pro (GP), Derived from Hibiscus sabdariffa, Exhibits Potent Antifibrotic Effects by Regulating the TGF-β1-ATF4-Serine/Glycine Biosynthesis Pathway

“…Both RT-qPCR and Western blot analysis demonstrated that GP inhibited ATF4 expression at the mRNA and protein levels, as shown in Figure 4a,b. Moreover, upon TGF-β1 stimulation, ATF4 undergoes nuclear translocation and then modulates the transcription of its target genes [5,23]. In the ICC analyses, TGF-β1 significantly increased the expression of ATF4 in the nucleus, and GP markedly reduced its expression (Figure 4c).…”

Deductive Toxicology and Bioorganometallics

“…14. Intracellular Signal Pathways That Mediate the Induction of CSE and CBS in Vascular Endothelial Cells[91][92][93] Supersulfides (reactive sulfur species) are produced by supersulfide-producing enzymes such as CSE and CBS and promote the proliferation of vascular endothelial cells 88). CSE and CBS are induced by activating the signaling pathways, including TGF-β-activated ALK5-Smad2/3/4 and ALK5-Smad2/3-ATF4; FGF-2-activated ERK1/2 pathway; and Cu10-activated ERK1/2, p38 MAPK, and HIF-1α/1β pathways.…”