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An In-Silico, In-Vitro and In-Vivo Combined Approach to Identify NMNATs as Potential Protein Targets of ProEGCG for Treatment of Endometriosis

Endometriosis is defined as endometrial tissues found outside the uterine cavity. ProEGCG is a prodrug of Epigallocatechin gallate (EGCG), a potent polyphenol found in green tea. It inhibits the development of endometriotic lesions of mouse model in vivo, with higher efficacy and more remarkable ant...

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Autores principales: Hung, Sze Wan, Liang, Bo, Gao, Yating, Zhang, Ruizhe, Tan, Zhouyurong, Zhang, Tao, Chung, Pui Wah Jacqueline, Chan, Tak Hang, Wang, Chi Chiu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8552031/
https://www.ncbi.nlm.nih.gov/pubmed/34721014
http://dx.doi.org/10.3389/fphar.2021.714790
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author Hung, Sze Wan
Liang, Bo
Gao, Yating
Zhang, Ruizhe
Tan, Zhouyurong
Zhang, Tao
Chung, Pui Wah Jacqueline
Chan, Tak Hang
Wang, Chi Chiu
author_facet Hung, Sze Wan
Liang, Bo
Gao, Yating
Zhang, Ruizhe
Tan, Zhouyurong
Zhang, Tao
Chung, Pui Wah Jacqueline
Chan, Tak Hang
Wang, Chi Chiu
author_sort Hung, Sze Wan
collection PubMed
description Endometriosis is defined as endometrial tissues found outside the uterine cavity. ProEGCG is a prodrug of Epigallocatechin gallate (EGCG), a potent polyphenol found in green tea. It inhibits the development of endometriotic lesions of mouse model in vivo, with higher efficacy and more remarkable anti-oxidative ability than EGCG. Our study aims to identify the molecular binding targets and pharmacological actions of ProEGCG in treating endometriosis. Protein target interaction study is essential to fully characterize the mechanism of actions, related therapeutic effects, and side effects. We employed a combined approach, starting with an in silico reverse screening of protein targets and molecular docking, followed by in vitro cellular thermal shift assay (CESTA) to assess the stability of protein-small molecule complexes. Then microarray and immunostaining of endometriotic lesions in mice in vivo confirmed the molecular interaction of the selected targets after treatment. Our study identified enzymes nicotinamide nucleotide adenylyltransferase (NMNAT)1 and NMNAT3 as protein targets of ProEGCG in silico and in vitro and were overexpressed after ProEGCG treatment in vivo. These findings suggested that participation in nicotinate and nicotinamide metabolism potentially regulated the redox status of endometriosis via its antioxidative capacities through binding to the potential therapeutic targets of ProEGCG.
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spelling pubmed-85520312021-10-29 An In-Silico, In-Vitro and In-Vivo Combined Approach to Identify NMNATs as Potential Protein Targets of ProEGCG for Treatment of Endometriosis Hung, Sze Wan Liang, Bo Gao, Yating Zhang, Ruizhe Tan, Zhouyurong Zhang, Tao Chung, Pui Wah Jacqueline Chan, Tak Hang Wang, Chi Chiu Front Pharmacol Pharmacology Endometriosis is defined as endometrial tissues found outside the uterine cavity. ProEGCG is a prodrug of Epigallocatechin gallate (EGCG), a potent polyphenol found in green tea. It inhibits the development of endometriotic lesions of mouse model in vivo, with higher efficacy and more remarkable anti-oxidative ability than EGCG. Our study aims to identify the molecular binding targets and pharmacological actions of ProEGCG in treating endometriosis. Protein target interaction study is essential to fully characterize the mechanism of actions, related therapeutic effects, and side effects. We employed a combined approach, starting with an in silico reverse screening of protein targets and molecular docking, followed by in vitro cellular thermal shift assay (CESTA) to assess the stability of protein-small molecule complexes. Then microarray and immunostaining of endometriotic lesions in mice in vivo confirmed the molecular interaction of the selected targets after treatment. Our study identified enzymes nicotinamide nucleotide adenylyltransferase (NMNAT)1 and NMNAT3 as protein targets of ProEGCG in silico and in vitro and were overexpressed after ProEGCG treatment in vivo. These findings suggested that participation in nicotinate and nicotinamide metabolism potentially regulated the redox status of endometriosis via its antioxidative capacities through binding to the potential therapeutic targets of ProEGCG. Frontiers Media S.A. 2021-10-14 /pmc/articles/PMC8552031/ /pubmed/34721014 http://dx.doi.org/10.3389/fphar.2021.714790 Text en Copyright © 2021 Hung, Liang, Gao, Zhang, Tan, Zhang, Chung, Chan and Wang. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Pharmacology
Hung, Sze Wan
Liang, Bo
Gao, Yating
Zhang, Ruizhe
Tan, Zhouyurong
Zhang, Tao
Chung, Pui Wah Jacqueline
Chan, Tak Hang
Wang, Chi Chiu
An In-Silico, In-Vitro and In-Vivo Combined Approach to Identify NMNATs as Potential Protein Targets of ProEGCG for Treatment of Endometriosis
title An In-Silico, In-Vitro and In-Vivo Combined Approach to Identify NMNATs as Potential Protein Targets of ProEGCG for Treatment of Endometriosis
title_full An In-Silico, In-Vitro and In-Vivo Combined Approach to Identify NMNATs as Potential Protein Targets of ProEGCG for Treatment of Endometriosis
title_fullStr An In-Silico, In-Vitro and In-Vivo Combined Approach to Identify NMNATs as Potential Protein Targets of ProEGCG for Treatment of Endometriosis
title_full_unstemmed An In-Silico, In-Vitro and In-Vivo Combined Approach to Identify NMNATs as Potential Protein Targets of ProEGCG for Treatment of Endometriosis
title_short An In-Silico, In-Vitro and In-Vivo Combined Approach to Identify NMNATs as Potential Protein Targets of ProEGCG for Treatment of Endometriosis
title_sort in-silico, in-vitro and in-vivo combined approach to identify nmnats as potential protein targets of proegcg for treatment of endometriosis
topic Pharmacology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8552031/
https://www.ncbi.nlm.nih.gov/pubmed/34721014
http://dx.doi.org/10.3389/fphar.2021.714790
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