Empagliflozin Attenuates Non-Alcoholic Fatty Liver Disease (NAFLD) in High Fat Diet Fed ApoE<sup>(-/-)</sup> Mice by Activating Autophagy and Reducing ER Stress and Apoptosis
Aims/hypothesis: SGLT-2 inhibitors (SGLT-2i) have been studied as potential treatments against NAFLD, showing varying beneficial effects. The molecular mechanisms mediating these effects have not been fully clarified. Herein, we investigated the impact of empagliflozin on NAFLD, focusing particularl...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-01-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/2/818 |
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| author | Narjes Nasiri-Ansari Chrysa Nikolopoulou Katerina Papoutsi Ioannis Kyrou Christos S. Mantzoros Georgios Kyriakopoulos Antonios Chatzigeorgiou Vassiliki Kalotychou Manpal S. Randeva Kamaljit Chatha Konstantinos Kontzoglou Gregory Kaltsas Athanasios G. Papavassiliou Harpal S. Randeva Eva Kassi |
| author_facet | Narjes Nasiri-Ansari Chrysa Nikolopoulou Katerina Papoutsi Ioannis Kyrou Christos S. Mantzoros Georgios Kyriakopoulos Antonios Chatzigeorgiou Vassiliki Kalotychou Manpal S. Randeva Kamaljit Chatha Konstantinos Kontzoglou Gregory Kaltsas Athanasios G. Papavassiliou Harpal S. Randeva Eva Kassi |
| author_sort | Narjes Nasiri-Ansari |
| collection | DOAJ |
| description | Aims/hypothesis: SGLT-2 inhibitors (SGLT-2i) have been studied as potential treatments against NAFLD, showing varying beneficial effects. The molecular mechanisms mediating these effects have not been fully clarified. Herein, we investigated the impact of empagliflozin on NAFLD, focusing particularly on ER stress, autophagy and apoptosis. Methods: Five-week old ApoE<sup>(-/-)</sup> mice were switched from normal to a high-fat diet (HFD). After five weeks, mice were randomly allocated into a control group (HFD + vehicle) and Empa group (HFD + empagliflozin 10 mg/kg/day) for five weeks. At the end of treatment, histomorphometric analysis was performed in liver, mRNA levels of <i>Fasn</i>, <i>Screbp-1</i>, <i>Scd-1</i>, <i>Ppar-γ</i>, <i>Pck-1</i>, <i>Mcp-1</i>, <i>Tnf-α</i>, <i>Il-6</i>, <i>F4/80</i>, <i>Atf4</i>, <i>Elf2α</i>, <i>Chop</i>, <i>Grp78</i>, <i>Grp94</i>, <i>Χbp1</i>, <i>Ire1α</i>, <i>Atf6</i>, <i>mTor</i>, <i>Lc3b</i>, <i>Beclin-1</i>, <i>P62</i>, <i>Bcl-2</i> and <i>Bax</i> were measured by qRT-PCR, and protein levels of p-EIF2α, EIF2a, CHOP, LC3II, P62, BECLIN-1 and cleaved CASPASE-8 were assessed by immunoblotting. Results: Empagliflozin-treated mice exhibited reduced fasting glucose, total cholesterol and triglyceride serum levels, as well as decreased NAFLD activity score, decreased expression of lipogenic enzymes (<i>Fasn</i>, <i>Screbp-1c</i> and <i>Pck-1</i>) and inflammatory molecules (<i>Mcp-1</i> and <i>F4/80</i>), compared to the Control group. Empagliflozin significantly decreased the expression of ER stress molecules <i>Grp78</i>, <i>Ire1α</i>, <i>Xbp1</i>, <i>Elf2α</i>, <i>Atf4</i>, <i>Atf6</i>, <i>Chop</i>, <i>P62(Sqstm1)</i> and <i>Grp94</i>; whilst activating autophagy via increased AMPK phosphorylation, decreased <i>mTOR</i> and increased <i>LC3B</i> expression. Finally, empagliflozin increased the <i>Bcl2/Bax</i> ratio and inhibited CASPASE-8 cleavage, reducing liver cell apoptosis. Immunoblotting analysis confirmed the qPCR results. Conclusion: These novel findings indicate that empagliflozin treatment for five weeks attenuates NAFLD progression in ApoE<sup>(-/-)</sup> mice by promoting autophagy, reducing ER stress and inhibiting hepatic apoptosis. |
| first_indexed | 2024-03-09T04:40:33Z |
| format | Article |
| id | doaj.art-f967148ee656483cac3dae9949ce40ff |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T04:40:33Z |
| publishDate | 2021-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-f967148ee656483cac3dae9949ce40ff2023-12-03T13:22:08ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-01-0122281810.3390/ijms22020818Empagliflozin Attenuates Non-Alcoholic Fatty Liver Disease (NAFLD) in High Fat Diet Fed ApoE<sup>(-/-)</sup> Mice by Activating Autophagy and Reducing ER Stress and ApoptosisNarjes Nasiri-Ansari0Chrysa Nikolopoulou1Katerina Papoutsi2Ioannis Kyrou3Christos S. Mantzoros4Georgios Kyriakopoulos5Antonios Chatzigeorgiou6Vassiliki Kalotychou7Manpal S. Randeva8Kamaljit Chatha9Konstantinos Kontzoglou10Gregory Kaltsas11Athanasios G. Papavassiliou12Harpal S. Randeva13Eva Kassi14Department of Biological Chemistry, Medical School, National and Kapodistrian University of Athens, 11527 Athens, GreeceDepartment of Biological Chemistry, Medical School, National and Kapodistrian University of Athens, 11527 Athens, GreeceDepartment of Biological Chemistry, Medical School, National and Kapodistrian University of Athens, 11527 Athens, GreeceWarwickshire Institute for the Study of Diabetes, Endocrinology and Metabolism (WISDEM), University Hospitals Coventry and Warwickshire NHS Trust, Coventry CV2 2DX, UKDivision of Endocrinology, Diabetes and Metabolism, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 02215, USADepartment of Biological Chemistry, Medical School, National and Kapodistrian University of Athens, 11527 Athens, GreeceDepartment of Physiology, Medical School, National and Kapodistrian University of Athens, 11527 Athens, Greece1st Department of Internal Medicine, Laiko Hospital, Medical School, National and Kapodistrian University of Athens, 11527 Athens, GreeceHuman Metabolism Research Unit, WISDEM Centre, NHS Trust, Coventry CV2 2DX, UKDepartment of Biochemistry & Immunology, University Hospitals Coventry and Warwickshire NHS Trust, Coventry CV2 2DX, UKLaboratory of Experimental Surgery and Surgical Research N.S. Christeas, Athens University Medical School, National and Kapodistrian University of Athens, 11527 Athens, GreeceEndocrine Oncology Unit, 1st Department of Propaupedic Internal Medicine, Laiko Hospital, National and Kapodistrian University of Athens, 11527 Athens, GreeceDepartment of Biological Chemistry, Medical School, National and Kapodistrian University of Athens, 11527 Athens, GreeceWarwickshire Institute for the Study of Diabetes, Endocrinology and Metabolism (WISDEM), University Hospitals Coventry and Warwickshire NHS Trust, Coventry CV2 2DX, UKDepartment of Biological Chemistry, Medical School, National and Kapodistrian University of Athens, 11527 Athens, GreeceAims/hypothesis: SGLT-2 inhibitors (SGLT-2i) have been studied as potential treatments against NAFLD, showing varying beneficial effects. The molecular mechanisms mediating these effects have not been fully clarified. Herein, we investigated the impact of empagliflozin on NAFLD, focusing particularly on ER stress, autophagy and apoptosis. Methods: Five-week old ApoE<sup>(-/-)</sup> mice were switched from normal to a high-fat diet (HFD). After five weeks, mice were randomly allocated into a control group (HFD + vehicle) and Empa group (HFD + empagliflozin 10 mg/kg/day) for five weeks. At the end of treatment, histomorphometric analysis was performed in liver, mRNA levels of <i>Fasn</i>, <i>Screbp-1</i>, <i>Scd-1</i>, <i>Ppar-γ</i>, <i>Pck-1</i>, <i>Mcp-1</i>, <i>Tnf-α</i>, <i>Il-6</i>, <i>F4/80</i>, <i>Atf4</i>, <i>Elf2α</i>, <i>Chop</i>, <i>Grp78</i>, <i>Grp94</i>, <i>Χbp1</i>, <i>Ire1α</i>, <i>Atf6</i>, <i>mTor</i>, <i>Lc3b</i>, <i>Beclin-1</i>, <i>P62</i>, <i>Bcl-2</i> and <i>Bax</i> were measured by qRT-PCR, and protein levels of p-EIF2α, EIF2a, CHOP, LC3II, P62, BECLIN-1 and cleaved CASPASE-8 were assessed by immunoblotting. Results: Empagliflozin-treated mice exhibited reduced fasting glucose, total cholesterol and triglyceride serum levels, as well as decreased NAFLD activity score, decreased expression of lipogenic enzymes (<i>Fasn</i>, <i>Screbp-1c</i> and <i>Pck-1</i>) and inflammatory molecules (<i>Mcp-1</i> and <i>F4/80</i>), compared to the Control group. Empagliflozin significantly decreased the expression of ER stress molecules <i>Grp78</i>, <i>Ire1α</i>, <i>Xbp1</i>, <i>Elf2α</i>, <i>Atf4</i>, <i>Atf6</i>, <i>Chop</i>, <i>P62(Sqstm1)</i> and <i>Grp94</i>; whilst activating autophagy via increased AMPK phosphorylation, decreased <i>mTOR</i> and increased <i>LC3B</i> expression. Finally, empagliflozin increased the <i>Bcl2/Bax</i> ratio and inhibited CASPASE-8 cleavage, reducing liver cell apoptosis. Immunoblotting analysis confirmed the qPCR results. Conclusion: These novel findings indicate that empagliflozin treatment for five weeks attenuates NAFLD progression in ApoE<sup>(-/-)</sup> mice by promoting autophagy, reducing ER stress and inhibiting hepatic apoptosis.https://www.mdpi.com/1422-0067/22/2/818NAFLDSGLT-2 inhibitorsautophagyER stressapoptosisinflammation |
| spellingShingle | Narjes Nasiri-Ansari Chrysa Nikolopoulou Katerina Papoutsi Ioannis Kyrou Christos S. Mantzoros Georgios Kyriakopoulos Antonios Chatzigeorgiou Vassiliki Kalotychou Manpal S. Randeva Kamaljit Chatha Konstantinos Kontzoglou Gregory Kaltsas Athanasios G. Papavassiliou Harpal S. Randeva Eva Kassi Empagliflozin Attenuates Non-Alcoholic Fatty Liver Disease (NAFLD) in High Fat Diet Fed ApoE<sup>(-/-)</sup> Mice by Activating Autophagy and Reducing ER Stress and Apoptosis International Journal of Molecular Sciences NAFLD SGLT-2 inhibitors autophagy ER stress apoptosis inflammation |
| title | Empagliflozin Attenuates Non-Alcoholic Fatty Liver Disease (NAFLD) in High Fat Diet Fed ApoE<sup>(-/-)</sup> Mice by Activating Autophagy and Reducing ER Stress and Apoptosis |
| title_full | Empagliflozin Attenuates Non-Alcoholic Fatty Liver Disease (NAFLD) in High Fat Diet Fed ApoE<sup>(-/-)</sup> Mice by Activating Autophagy and Reducing ER Stress and Apoptosis |
| title_fullStr | Empagliflozin Attenuates Non-Alcoholic Fatty Liver Disease (NAFLD) in High Fat Diet Fed ApoE<sup>(-/-)</sup> Mice by Activating Autophagy and Reducing ER Stress and Apoptosis |
| title_full_unstemmed | Empagliflozin Attenuates Non-Alcoholic Fatty Liver Disease (NAFLD) in High Fat Diet Fed ApoE<sup>(-/-)</sup> Mice by Activating Autophagy and Reducing ER Stress and Apoptosis |
| title_short | Empagliflozin Attenuates Non-Alcoholic Fatty Liver Disease (NAFLD) in High Fat Diet Fed ApoE<sup>(-/-)</sup> Mice by Activating Autophagy and Reducing ER Stress and Apoptosis |
| title_sort | empagliflozin attenuates non alcoholic fatty liver disease nafld in high fat diet fed apoe sup sup mice by activating autophagy and reducing er stress and apoptosis |
| topic | NAFLD SGLT-2 inhibitors autophagy ER stress apoptosis inflammation |
| url | https://www.mdpi.com/1422-0067/22/2/818 |
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