Chenodeoxycholic Acid Has Non-Thermogenic, Mitodynamic Anti-Obesity Effects in an In Vitro CRISPR/Cas9 Model of Bile Acid Receptor TGR5 Knockdown

Bile acids (BA) have shown promising effects in animal models of obesity. However, the said effects are thought to rely on a thermogenic effect, which is questionably present in humans. A previous work has shown that the BA chenodeoxycholic acid (CDCA) can revert obesity and accelerate metabolism in...

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Autores principales:	João S. Teodoro, Ivo F. Machado, Ana C. Castela, João A. Amorim, Ivana Jarak, Rui A. Carvalho, Carlos M. Palmeira, Anabela P. Rolo
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	CDCA TGR5 mitochondria mitophagy CRISPR/Cas9 3T3-L1 Biology (General) QH301-705.5 Chemistry QD1-999
Acceso en línea:	https://doaj.org/article/76f07e45d49847118408fbd5a0625b53
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spelling	oai:doaj.org-article:76f07e45d49847118408fbd5a0625b532021-11-11T17:11:48ZChenodeoxycholic Acid Has Non-Thermogenic, Mitodynamic Anti-Obesity Effects in an In Vitro CRISPR/Cas9 Model of Bile Acid Receptor TGR5 Knockdown10.3390/ijms2221117381422-00671661-6596https://doaj.org/article/76f07e45d49847118408fbd5a0625b532021-10-01T00:00:00Zhttps://www.mdpi.com/1422-0067/22/21/11738https://doaj.org/toc/1661-6596https://doaj.org/toc/1422-0067Bile acids (BA) have shown promising effects in animal models of obesity. However, the said effects are thought to rely on a thermogenic effect, which is questionably present in humans. A previous work has shown that the BA chenodeoxycholic acid (CDCA) can revert obesity and accelerate metabolism in animal and cell culture models. Thus, the aim of this study was to understand if this obesity reduction is indeed thermogenically-dependent. A CRISPR/Cas9 model of TGR5 (BA receptor) knockdown in 3T3-L1 adipocytes was developed to diminish thermogenic effects. Various parameters were assessed, including mitochondrial bioenergetics by Seahorse flux analysis, oxidative stress and membrane potential by fluorometry, intermediary metabolism by NMR, protein content assessment by Western Blot, gene expression by qPCR, and confocal microscopy evaluation of mitophagy. CDCA was still capable, for the most part, of reversing the harmful effects of cellular obesity, elevating mitophagy and leading to the reduction of harmed mitochondria within the cells, boosting mitochondrial activity, and thus energy consumption. In summary, CDCA has a non-thermogenic, obesity reducing capacity that hinges on a healthy mitochondrial population, explaining at least some of these effects and opening avenues of human treatment for metabolic diseases.João S. TeodoroIvo F. MachadoAna C. CastelaJoão A. AmorimIvana JarakRui A. CarvalhoCarlos M. PalmeiraAnabela P. RoloMDPI AGarticleCDCATGR5mitochondriamitophagyCRISPR/Cas93T3-L1Biology (General)QH301-705.5ChemistryQD1-999ENInternational Journal of Molecular Sciences, Vol 22, Iss 11738, p 11738 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	CDCA TGR5 mitochondria mitophagy CRISPR/Cas9 3T3-L1 Biology (General) QH301-705.5 Chemistry QD1-999
spellingShingle	CDCA TGR5 mitochondria mitophagy CRISPR/Cas9 3T3-L1 Biology (General) QH301-705.5 Chemistry QD1-999 João S. Teodoro Ivo F. Machado Ana C. Castela João A. Amorim Ivana Jarak Rui A. Carvalho Carlos M. Palmeira Anabela P. Rolo Chenodeoxycholic Acid Has Non-Thermogenic, Mitodynamic Anti-Obesity Effects in an In Vitro CRISPR/Cas9 Model of Bile Acid Receptor TGR5 Knockdown
description	Bile acids (BA) have shown promising effects in animal models of obesity. However, the said effects are thought to rely on a thermogenic effect, which is questionably present in humans. A previous work has shown that the BA chenodeoxycholic acid (CDCA) can revert obesity and accelerate metabolism in animal and cell culture models. Thus, the aim of this study was to understand if this obesity reduction is indeed thermogenically-dependent. A CRISPR/Cas9 model of TGR5 (BA receptor) knockdown in 3T3-L1 adipocytes was developed to diminish thermogenic effects. Various parameters were assessed, including mitochondrial bioenergetics by Seahorse flux analysis, oxidative stress and membrane potential by fluorometry, intermediary metabolism by NMR, protein content assessment by Western Blot, gene expression by qPCR, and confocal microscopy evaluation of mitophagy. CDCA was still capable, for the most part, of reversing the harmful effects of cellular obesity, elevating mitophagy and leading to the reduction of harmed mitochondria within the cells, boosting mitochondrial activity, and thus energy consumption. In summary, CDCA has a non-thermogenic, obesity reducing capacity that hinges on a healthy mitochondrial population, explaining at least some of these effects and opening avenues of human treatment for metabolic diseases.
format	article
author	João S. Teodoro Ivo F. Machado Ana C. Castela João A. Amorim Ivana Jarak Rui A. Carvalho Carlos M. Palmeira Anabela P. Rolo
author_facet	João S. Teodoro Ivo F. Machado Ana C. Castela João A. Amorim Ivana Jarak Rui A. Carvalho Carlos M. Palmeira Anabela P. Rolo
author_sort	João S. Teodoro
title	Chenodeoxycholic Acid Has Non-Thermogenic, Mitodynamic Anti-Obesity Effects in an In Vitro CRISPR/Cas9 Model of Bile Acid Receptor TGR5 Knockdown
title_short	Chenodeoxycholic Acid Has Non-Thermogenic, Mitodynamic Anti-Obesity Effects in an In Vitro CRISPR/Cas9 Model of Bile Acid Receptor TGR5 Knockdown
title_full	Chenodeoxycholic Acid Has Non-Thermogenic, Mitodynamic Anti-Obesity Effects in an In Vitro CRISPR/Cas9 Model of Bile Acid Receptor TGR5 Knockdown
title_fullStr	Chenodeoxycholic Acid Has Non-Thermogenic, Mitodynamic Anti-Obesity Effects in an In Vitro CRISPR/Cas9 Model of Bile Acid Receptor TGR5 Knockdown
title_full_unstemmed	Chenodeoxycholic Acid Has Non-Thermogenic, Mitodynamic Anti-Obesity Effects in an In Vitro CRISPR/Cas9 Model of Bile Acid Receptor TGR5 Knockdown
title_sort	chenodeoxycholic acid has non-thermogenic, mitodynamic anti-obesity effects in an in vitro crispr/cas9 model of bile acid receptor tgr5 knockdown
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/76f07e45d49847118408fbd5a0625b53
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Chenodeoxycholic Acid Has Non-Thermogenic, Mitodynamic Anti-Obesity Effects in an In Vitro CRISPR/Cas9 Model of Bile Acid Receptor TGR5 Knockdown

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