The selective blockade of metabotropic glutamate receptor-5 attenuates fat accumulation in an <em>in vitro</em> model of benign steatosis
It has been previously found that the blockade of metabotropic glutamate receptor type 5 (mGluR5) protects against hepatic ischemia/reperfusion injury and acetaminophen toxicity. The role of mGluR5 in NAFLD has not yet been elucidated. Here, we evaluated the effects of mGluR5 blockade in an in vitro...
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| Format: | Article |
| Language: | English |
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PAGEPress Publications
2020-11-01
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| Series: | European Journal of Histochemistry |
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| Online Access: | https://ejh.it/index.php/ejh/article/view/3175 |
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| author | Andrea Ferrigno Clarissa Berardo Laura Giuseppina Di Pasqua Marta Cagna Veronica Siciliano Plinio Richelmi Mariapia Vairetti |
| author_facet | Andrea Ferrigno Clarissa Berardo Laura Giuseppina Di Pasqua Marta Cagna Veronica Siciliano Plinio Richelmi Mariapia Vairetti |
| author_sort | Andrea Ferrigno |
| collection | DOAJ |
| description | It has been previously found that the blockade of metabotropic glutamate receptor type 5 (mGluR5) protects against hepatic ischemia/reperfusion injury and acetaminophen toxicity. The role of mGluR5 in NAFLD has not yet been elucidated. Here, we evaluated the effects of mGluR5 blockade in an in vitro model of steatosis. HepG2 cells were pre-incubated for 12 h with an mGluR5 agonist, a negative allosteric modulator (DHPG and MPEP, respectively) or vehicle, then treated with 1.5 mM oleate/palmitate (O/P) for another 12 h. Cell viability was evaluated with the MTT assay; fat accumulation was measured using the fluorescent dye nile red; SREBP-1, PPAR-α, iNOS and Caspase-3 protein expression were evaluated by Western blot; NFkB activity was evaluated as pNFkB/NFkB ratio. mGluR5 modulation did not alter cell viability in O/P-incubated cells; MPEP prevented intracellular lipid accumulation in O/P treated cells; MPEP administration was also associated with a reversion of O/P-induced changes in SREBP-1 and PPAR-α expression, involved in free fatty acid (FFA) metabolism and uptake. No changes were observed in iNOS and Caspase-3 expression, or in NFkB activity. In conclusion, mGluR5 pharmacological blockade reduced fat accumulation in HepG2 cells incubated with O/P, probably by modulating the expression of SREBP-1 and PPAR-α. |
| first_indexed | 2024-12-11T07:19:29Z |
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| id | doaj.art-0afdce3f76d0428d83c21a3873d2d485 |
| institution | Directory Open Access Journal |
| issn | 1121-760X 2038-8306 |
| language | English |
| last_indexed | 2024-12-11T07:19:29Z |
| publishDate | 2020-11-01 |
| publisher | PAGEPress Publications |
| record_format | Article |
| series | European Journal of Histochemistry |
| spelling | doaj.art-0afdce3f76d0428d83c21a3873d2d4852022-12-22T01:16:08ZengPAGEPress PublicationsEuropean Journal of Histochemistry1121-760X2038-83062020-11-0164410.4081/ejh.2020.3175The selective blockade of metabotropic glutamate receptor-5 attenuates fat accumulation in an <em>in vitro</em> model of benign steatosisAndrea Ferrigno0Clarissa Berardo1Laura Giuseppina Di Pasqua2Marta Cagna3Veronica Siciliano4Plinio Richelmi5Mariapia Vairetti6Department of Internal Medicine and Therapeutics, Unit of Cellular and Molecular Pharmacology and Toxicology, University of PaviaDepartment of Internal Medicine and Therapeutics, Unit of Cellular and Molecular Pharmacology and Toxicology, University of PaviaDepartment of Internal Medicine and Therapeutics, Unit of Cellular and Molecular Pharmacology and Toxicology, University of PaviaDepartment of Internal Medicine and Therapeutics, Unit of Cellular and Molecular Pharmacology and Toxicology, University of PaviaDepartment of Internal Medicine and Therapeutics, Unit of Cellular and Molecular Pharmacology and Toxicology, University of PaviaDepartment of Internal Medicine and Therapeutics, Unit of Cellular and Molecular Pharmacology and Toxicology, University of PaviaDepartment of Internal Medicine and Therapeutics, Unit of Cellular and Molecular Pharmacology and Toxicology, University of PaviaIt has been previously found that the blockade of metabotropic glutamate receptor type 5 (mGluR5) protects against hepatic ischemia/reperfusion injury and acetaminophen toxicity. The role of mGluR5 in NAFLD has not yet been elucidated. Here, we evaluated the effects of mGluR5 blockade in an in vitro model of steatosis. HepG2 cells were pre-incubated for 12 h with an mGluR5 agonist, a negative allosteric modulator (DHPG and MPEP, respectively) or vehicle, then treated with 1.5 mM oleate/palmitate (O/P) for another 12 h. Cell viability was evaluated with the MTT assay; fat accumulation was measured using the fluorescent dye nile red; SREBP-1, PPAR-α, iNOS and Caspase-3 protein expression were evaluated by Western blot; NFkB activity was evaluated as pNFkB/NFkB ratio. mGluR5 modulation did not alter cell viability in O/P-incubated cells; MPEP prevented intracellular lipid accumulation in O/P treated cells; MPEP administration was also associated with a reversion of O/P-induced changes in SREBP-1 and PPAR-α expression, involved in free fatty acid (FFA) metabolism and uptake. No changes were observed in iNOS and Caspase-3 expression, or in NFkB activity. In conclusion, mGluR5 pharmacological blockade reduced fat accumulation in HepG2 cells incubated with O/P, probably by modulating the expression of SREBP-1 and PPAR-α.https://ejh.it/index.php/ejh/article/view/3175NAFLDmGluR5MPEPoleate/palmitateHepG2SREBP-1 |
| spellingShingle | Andrea Ferrigno Clarissa Berardo Laura Giuseppina Di Pasqua Marta Cagna Veronica Siciliano Plinio Richelmi Mariapia Vairetti The selective blockade of metabotropic glutamate receptor-5 attenuates fat accumulation in an <em>in vitro</em> model of benign steatosis European Journal of Histochemistry NAFLD mGluR5 MPEP oleate/palmitate HepG2 SREBP-1 |
| title | The selective blockade of metabotropic glutamate receptor-5 attenuates fat accumulation in an <em>in vitro</em> model of benign steatosis |
| title_full | The selective blockade of metabotropic glutamate receptor-5 attenuates fat accumulation in an <em>in vitro</em> model of benign steatosis |
| title_fullStr | The selective blockade of metabotropic glutamate receptor-5 attenuates fat accumulation in an <em>in vitro</em> model of benign steatosis |
| title_full_unstemmed | The selective blockade of metabotropic glutamate receptor-5 attenuates fat accumulation in an <em>in vitro</em> model of benign steatosis |
| title_short | The selective blockade of metabotropic glutamate receptor-5 attenuates fat accumulation in an <em>in vitro</em> model of benign steatosis |
| title_sort | selective blockade of metabotropic glutamate receptor 5 attenuates fat accumulation in an em in vitro em model of benign steatosis |
| topic | NAFLD mGluR5 MPEP oleate/palmitate HepG2 SREBP-1 |
| url | https://ejh.it/index.php/ejh/article/view/3175 |
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