Characterization of zofenoprilat as an inducer of functional angiogenesis through increased H₂S availability

Abstract

Background and Purpose Hydrogen sulfide (H2S), an endogenous volatile mediator with pleiotropic functions, promotes vasorelaxation, exerts anti‐inflammatory actions and regulates angiogenesis. Previously, the SH‐containing angiotensin‐converting enzyme inhibitor (ACEI), zofenopril, was identified as being effective in preserving endothelial function and inducing angiogenesis among ACEIs. Based on the H2S donor property of its active metabolite zofenoprilat, the objective of this study was to evaluate whether zofenoprilat‐induced angiogenesis was due to increased H2S availability. Experimental Approach HUVECs were used for in vitro studies of angiogenesis, whereas the Matrigel plug assay was used for in vivo assessments. Key Results Zofenoprilat‐treated HUVECs showed an increase in all functional features of the angiogenic process in vitro. As zofenoprilat induced the expression of CSE (cystathionine‐γ‐lyase) and the continuous production of H2S, CSE inhibition or silencing blocked the ability of zofenoprilat to induce angiogenesis, both in vitro and in vivo. The molecular mechanisms underlying H2S/zofenoprilat‐induced angiogenesis were dependent on Akt, eNOS and ERK1/2 cascades. ATP‐sensitive potassium (KATP) channels, the molecular target that mediates part of the vascular functions of H2S, were shown to be involved in the upstream activation of Akt and ERK1/2. Moreover, the up‐regulation of fibroblast growth factor‐2 was dependent on CSE‐derived H2S response to H2S and KATP activation. Conclusions and Implications Zofenoprilat induced a constant production of H2S that stimulated the angiogenic process through a KATP channel/Akt/eNOS/ERK1/2 pathway. Thus, zofenopril can be considered as a pro‐angiogenic drug acting through H2S release and production, useful in cardiovascular pathologies where vascular functions need to be re‐established and functional angiogenesis induced.

Regulation and role of endogenously produced hydrogen sulfide in angiogenesis

“…In a follow up study, Terzuoli et al reported that the zofenoprilat-induced angiogenic properties of endothelial cells were attenuated by CSE inhibition or silencing [94]. Similarly, PAG administration reduced neovascularization in the Matrigel® plug assay in vivo [94]. The molecular mechanisms underlying zofenopril-induced angiogenesis were dependent on Akt, eNOS and ERK1/2 cascades, in line with what is known about mediators of H 2 S-triggered angiogenic signaling.…”

“…In a follow up study, Terzuoli et al reported that the zofenoprilat-induced angiogenic properties of endothelial cells were attenuated by CSE inhibition or silencing [94]. Similarly, PAG administration reduced neovascularization in the Matrigel® plug assay in vivo [94].…”

Comparison of the Effect of Two Hyaluronic Acid Preparations on Fibroblast and Endothelial Cell Functions Related to Angiogenesis

“…All experimental points using cells from the single culture plate were run in triplicate. After 2 days, cells were fixed, stained with H&E and randomly counted at 20× original magnification in 5 fields [19,20]. Data are reported as number of cells counted/well.…”

Targeting endothelial cell metabolism for cardio-protection from the toxicity of antitumor agents

“…We have recently reported that ACEi dependent H 2 S exerts protective activity on ECs through activation of K ATP channel, PI-3 K and eNOS, as well as up-regulation of FGF-2 and telomerase, thus maintaining endothelial longevity and function [76][77][78]104] (Fig. 4).…”

“…Zofenoprilat induces a constant H 2 S production through CSE upregulation in vascular endothelium. On its turn H 2 S maintains ECs survival and stimulates the angiogenic process through a sequential pathway involving K ATP channel/Akt/ eNOS/ERK1/2 pathway (for details see [104]) studies evaluated the effectiveness of common antioxidants found in the diet, which are known to prevent cellular oxidative damage. Many animal models showed a reduction in anthracycline cardiotoxicity by coadministering compounds such as vitamin E, vitamin C, vitamin A, coenzyme Q, and flavonoids.…”