Effects of Moringa oleifera Leaves Extract on High Glucose-Induced Metabolic Changes in HepG2 Cells
Mitochondrial dysfunction is a hallmark of diabetes, but the metabolic alterations during early stages of the disease remain unknown. The ability of liver cells to rearrange their metabolism plays an important role in compensating the energy shortage and may provide cell survival. Moringa oleifera l...
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2018-06-01
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author | Jorge A. Sosa-Gutiérrez Mónica A. Valdéz-Solana Tamara Y. Forbes-Hernández Claudia I. Avitia-Domínguez Gonzalo G. Garcia-Vargas José M. Salas-Pacheco Oscar Flores-Herrera Alfredo Téllez-Valencia Maurizio Battino Erick Sierra-Campos |
author_facet | Jorge A. Sosa-Gutiérrez Mónica A. Valdéz-Solana Tamara Y. Forbes-Hernández Claudia I. Avitia-Domínguez Gonzalo G. Garcia-Vargas José M. Salas-Pacheco Oscar Flores-Herrera Alfredo Téllez-Valencia Maurizio Battino Erick Sierra-Campos |
author_sort | Jorge A. Sosa-Gutiérrez |
collection | DOAJ |
description | Mitochondrial dysfunction is a hallmark of diabetes, but the metabolic alterations during early stages of the disease remain unknown. The ability of liver cells to rearrange their metabolism plays an important role in compensating the energy shortage and may provide cell survival. Moringa oleifera leaves have been studied for its health properties against diabetes, insulin resistance, and non-alcoholic liver disease. We postulated that M. oleifera executes a protective function on mitochondrial functionality in HepG2 treated with high glucose. We evaluated the effect of high glucose treatment on the mitochondrial function of HepG2 cells using a Seahorse extracellular flux analyzer (Agilent, Santa Clara, CA, USA), blue native polyacrylamide gel electrophoresis (BN-PAGE), and western blot analysis. For assessment of mitochondrial abnormalities, we measured the activity of mitochondrial Complex I and IV as well as uncoupling protein 2, and sirtuin 3 protein contents. Our results demonstrate that, under conditions mimicking the hyperglycemia, Complex I activity, UCP2, Complex III and IV subunits content, supercomplex formation, and acetylation levels are modified with respect to the control condition. However, basal oxygen consumption rate was not affected and mitochondrial reactive oxygen species production remained unchanged in all groups. Treatment of HepG2 cells with M. oleifera extract significantly increased both protein content and mitochondrial complexes activities. Nonetheless, control cells’ respiratory control ratio (RCR) was 4.37 compared to high glucose treated cells’ RCR of 15.3, and glucose plus M. oleifera treated cells’ RCR of 5.2, this indicates high-quality mitochondria and efficient oxidative phosphorylation coupling. Additionally, the state app was not altered between different treatments, suggesting no alteration in respiratory fluxes. These findings enhance understanding of the actions of M. oleifera and suggest that the known antidiabetic property of this plant, at least in part, is mediated through modulating the mitochondrial respiratory chain. |
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spelling | doaj.art-db09fc645cfa4afd8c70365a4e64c7d92023-08-02T07:16:40ZengMDPI AGBiology2079-77372018-06-01733710.3390/biology7030037biology7030037Effects of Moringa oleifera Leaves Extract on High Glucose-Induced Metabolic Changes in HepG2 CellsJorge A. Sosa-Gutiérrez0Mónica A. Valdéz-Solana1Tamara Y. Forbes-Hernández2Claudia I. Avitia-Domínguez3Gonzalo G. Garcia-Vargas4José M. Salas-Pacheco5Oscar Flores-Herrera6Alfredo Téllez-Valencia7Maurizio Battino8Erick Sierra-Campos9Facultad de Ciencias Químicas, Universidad Juárez del Estado de Durango Campus Gómez Palacio, Avenida Artículo 123 S/N, Fracc, Filadelfia, 35010 Gómez Palacio, MexicoFacultad de Ciencias Químicas, Universidad Juárez del Estado de Durango Campus Gómez Palacio, Avenida Artículo 123 S/N, Fracc, Filadelfia, 35010 Gómez Palacio, MexicoDipartimento di Scienze Cliniche Specialistiche ed Odontostomatologiche (DISCO)-Sez. Biochimica, Facoltà di Medicina, Università Politecnica delle Marche, 60131 Ancona, ItalyFacultad de Medicina y Nutrición, Universidad Juárez del Estado de Durango Campus Durango, Avenida Universidad y Fanny Anitúa S/N, 34000 Durango, MexicoFacultad de Ciencias de la Salud, Universidad Juárez del Estado de Durango Campus Gómez Palacio, Calzada Palmas 1, Colonia Revolución, 35050 Gómez Palacio, MexicoInstituto de Investigación Científica, Universidad Juárez del Estado de Durango, Avenida Universidad S/N, 34000 Durango, MexicoDepartamento de Bioquímica, Facultad de Medicina, Universidad Nacional Autónoma de México, 04510 Ciudad de México, MexicoFacultad de Medicina y Nutrición, Universidad Juárez del Estado de Durango Campus Durango, Avenida Universidad y Fanny Anitúa S/N, 34000 Durango, MexicoDipartimento di Scienze Cliniche Specialistiche ed Odontostomatologiche (DISCO)-Sez. Biochimica, Facoltà di Medicina, Università Politecnica delle Marche, 60131 Ancona, ItalyFacultad de Ciencias Químicas, Universidad Juárez del Estado de Durango Campus Gómez Palacio, Avenida Artículo 123 S/N, Fracc, Filadelfia, 35010 Gómez Palacio, MexicoMitochondrial dysfunction is a hallmark of diabetes, but the metabolic alterations during early stages of the disease remain unknown. The ability of liver cells to rearrange their metabolism plays an important role in compensating the energy shortage and may provide cell survival. Moringa oleifera leaves have been studied for its health properties against diabetes, insulin resistance, and non-alcoholic liver disease. We postulated that M. oleifera executes a protective function on mitochondrial functionality in HepG2 treated with high glucose. We evaluated the effect of high glucose treatment on the mitochondrial function of HepG2 cells using a Seahorse extracellular flux analyzer (Agilent, Santa Clara, CA, USA), blue native polyacrylamide gel electrophoresis (BN-PAGE), and western blot analysis. For assessment of mitochondrial abnormalities, we measured the activity of mitochondrial Complex I and IV as well as uncoupling protein 2, and sirtuin 3 protein contents. Our results demonstrate that, under conditions mimicking the hyperglycemia, Complex I activity, UCP2, Complex III and IV subunits content, supercomplex formation, and acetylation levels are modified with respect to the control condition. However, basal oxygen consumption rate was not affected and mitochondrial reactive oxygen species production remained unchanged in all groups. Treatment of HepG2 cells with M. oleifera extract significantly increased both protein content and mitochondrial complexes activities. Nonetheless, control cells’ respiratory control ratio (RCR) was 4.37 compared to high glucose treated cells’ RCR of 15.3, and glucose plus M. oleifera treated cells’ RCR of 5.2, this indicates high-quality mitochondria and efficient oxidative phosphorylation coupling. Additionally, the state app was not altered between different treatments, suggesting no alteration in respiratory fluxes. These findings enhance understanding of the actions of M. oleifera and suggest that the known antidiabetic property of this plant, at least in part, is mediated through modulating the mitochondrial respiratory chain.http://www.mdpi.com/2079-7737/7/3/37HepG2 cellsMoringa oleiferamitochondriaUCP2SIRT3 |
spellingShingle | Jorge A. Sosa-Gutiérrez Mónica A. Valdéz-Solana Tamara Y. Forbes-Hernández Claudia I. Avitia-Domínguez Gonzalo G. Garcia-Vargas José M. Salas-Pacheco Oscar Flores-Herrera Alfredo Téllez-Valencia Maurizio Battino Erick Sierra-Campos Effects of Moringa oleifera Leaves Extract on High Glucose-Induced Metabolic Changes in HepG2 Cells Biology HepG2 cells Moringa oleifera mitochondria UCP2 SIRT3 |
title | Effects of Moringa oleifera Leaves Extract on High Glucose-Induced Metabolic Changes in HepG2 Cells |
title_full | Effects of Moringa oleifera Leaves Extract on High Glucose-Induced Metabolic Changes in HepG2 Cells |
title_fullStr | Effects of Moringa oleifera Leaves Extract on High Glucose-Induced Metabolic Changes in HepG2 Cells |
title_full_unstemmed | Effects of Moringa oleifera Leaves Extract on High Glucose-Induced Metabolic Changes in HepG2 Cells |
title_short | Effects of Moringa oleifera Leaves Extract on High Glucose-Induced Metabolic Changes in HepG2 Cells |
title_sort | effects of moringa oleifera leaves extract on high glucose induced metabolic changes in hepg2 cells |
topic | HepG2 cells Moringa oleifera mitochondria UCP2 SIRT3 |
url | http://www.mdpi.com/2079-7737/7/3/37 |
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