Publicação
New insights on the inhibition of neutrophil extracellular traps enzymes in equine endometrium
| Resumo: | Mares physiologically develop a post-breeding endometritis characterized by a fast arrival of neutrophils into the uterine lumen. These neutrophils besides releasing granules of proteolytic and cytotoxic enzymes, may also deliver to the extracellular environment their DNA, histones and enzymes forming neutrophil extracellular traps (NETs). Besides trapping and fighting pathogens, NETs persistence has been also associated to the development of pathological conditions, such as fibrosis. The enzymes found in NETs, such as elastase (ELA), cathepsin G (CAT) and myeloperoxidase (MPO) act as pro-fibrotic factors in equine endometrial fibrosis, by inducing collagen type I (COL1) accumulation. Matrix metallopeptidases (MMPs) are crucial for this extracellular matrix remodeling. Prostaglandins (PG)s E2 and F2α have been described as possessing anti or pro-fibrotic effects. Equine endometrial explants from follicular phase (FP) or mid-luteal phase (MLP) were treated in vitro with ELA, CAT or MPO and their specific inhibitors for 24 or 48h. This work aimed to evaluate the explants response to: (i) ELA inhibition by sivelestat sodium salt (SIV) on COL1A2 transcription and PGE2 and PGF2α secretion; (ii) ELA and SIV treatment on MMP-2 and MMP-9 activity, and the inhibitory effect of SIV on ELA-induced COL1; (iii) CAT and Cathepsin G inhibitor I (β-keto-phosphonic acid; INH) treatment on MMP-2 and MMP-9 activity, and the effect of INH on CAT-induced COL1 production; (iv) the inhibitory effect of 4-aminobenzoic acid hydrazide (ABAH) on MPO-induced COL1 and the effect of MPO and ABAH on MMP-2 and MMP-9 gelatinolytic activity. In FP, COL1A2 transcription decreased in SIV-treated group, simultaneously with reduced pro-fibrotic PGF2α and increased anti-fibrotic PGE2 production. In ELA- and SIV-treated explants, MMPs expression depended on estrous cycle phase and time of treatment. Sivelestat inhibited ELA-induced COL1A2 transcripts in FP (24 h) and MLP (24 h, 48 h). The effect of INH was observed on CAT-induced COL1 in both phases at 48h. The MMP-2 might be involved in an earlier response to CAT, while MMP-9 in a later response in FP. The inhibitory effect of ABAH on MPO-induced COL1 was detected in FP at 48h. Matrix metallopeptidase-2 appears to be involved in an acute response to MPO treatment in MLP and MMP-9 in FP in a prolonged MPO treatment. The use of specific inhibitors of ELA, CAT or MPO, might be the grounds for future development of specific drugs to be used as prophylaxis or therapy of endometrosis in the mare. |
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| Autores principais: | Amaral, Ana Sofia Pires |
| Assunto: | Elastase cathepsin G myeloperoxidase inhibition endometrosis Elastase catepsina G myeloperoxidase inibição endometrose |
| Ano: | 2021 |
| País: | Portugal |
| Tipo de documento: | tese de doutoramento |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Universidade de Lisboa |
| Idioma: | inglês |
| Origem: | Repositório da Universidade de Lisboa |
| Resumo: | Mares physiologically develop a post-breeding endometritis characterized by a fast arrival of neutrophils into the uterine lumen. These neutrophils besides releasing granules of proteolytic and cytotoxic enzymes, may also deliver to the extracellular environment their DNA, histones and enzymes forming neutrophil extracellular traps (NETs). Besides trapping and fighting pathogens, NETs persistence has been also associated to the development of pathological conditions, such as fibrosis. The enzymes found in NETs, such as elastase (ELA), cathepsin G (CAT) and myeloperoxidase (MPO) act as pro-fibrotic factors in equine endometrial fibrosis, by inducing collagen type I (COL1) accumulation. Matrix metallopeptidases (MMPs) are crucial for this extracellular matrix remodeling. Prostaglandins (PG)s E2 and F2α have been described as possessing anti or pro-fibrotic effects. Equine endometrial explants from follicular phase (FP) or mid-luteal phase (MLP) were treated in vitro with ELA, CAT or MPO and their specific inhibitors for 24 or 48h. This work aimed to evaluate the explants response to: (i) ELA inhibition by sivelestat sodium salt (SIV) on COL1A2 transcription and PGE2 and PGF2α secretion; (ii) ELA and SIV treatment on MMP-2 and MMP-9 activity, and the inhibitory effect of SIV on ELA-induced COL1; (iii) CAT and Cathepsin G inhibitor I (β-keto-phosphonic acid; INH) treatment on MMP-2 and MMP-9 activity, and the effect of INH on CAT-induced COL1 production; (iv) the inhibitory effect of 4-aminobenzoic acid hydrazide (ABAH) on MPO-induced COL1 and the effect of MPO and ABAH on MMP-2 and MMP-9 gelatinolytic activity. In FP, COL1A2 transcription decreased in SIV-treated group, simultaneously with reduced pro-fibrotic PGF2α and increased anti-fibrotic PGE2 production. In ELA- and SIV-treated explants, MMPs expression depended on estrous cycle phase and time of treatment. Sivelestat inhibited ELA-induced COL1A2 transcripts in FP (24 h) and MLP (24 h, 48 h). The effect of INH was observed on CAT-induced COL1 in both phases at 48h. The MMP-2 might be involved in an earlier response to CAT, while MMP-9 in a later response in FP. The inhibitory effect of ABAH on MPO-induced COL1 was detected in FP at 48h. Matrix metallopeptidase-2 appears to be involved in an acute response to MPO treatment in MLP and MMP-9 in FP in a prolonged MPO treatment. The use of specific inhibitors of ELA, CAT or MPO, might be the grounds for future development of specific drugs to be used as prophylaxis or therapy of endometrosis in the mare. |
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