Glucolipotoxicity Suppressed Autophagy and Insulin Contents in Human Islets, and Attenuation of PERK Activity Enhanced Them in an ATG7-Dependent Manner
Background Administration of pancreatic endoplasmic reticulum kinase inhibitor (PERKi) improved insulin secretion and hyperglycemia in obese diabetic mice. In this study, autophagic balance was studied whether to mediate it. Methods Human islets were isolated from living patients without diabetes. P...
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| Format: | Article |
| Language: | English |
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Korean Diabetes Association
2024-03-01
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| Series: | Diabetes & Metabolism Journal |
| Subjects: | |
| Online Access: | http://e-dmj.org/upload/pdf/dmj-2022-0366.pdf |
| _version_ | 1797221647744237568 |
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| author | Seoil Moon Ji Yoon Lim Mirang Lee Youngmin Han Hongbeom Kim Wooil Kwon Jin-Young Jang Mi Na Kim Kyong Soo Park Hye Seung Jung |
| author_facet | Seoil Moon Ji Yoon Lim Mirang Lee Youngmin Han Hongbeom Kim Wooil Kwon Jin-Young Jang Mi Na Kim Kyong Soo Park Hye Seung Jung |
| author_sort | Seoil Moon |
| collection | DOAJ |
| description | Background Administration of pancreatic endoplasmic reticulum kinase inhibitor (PERKi) improved insulin secretion and hyperglycemia in obese diabetic mice. In this study, autophagic balance was studied whether to mediate it. Methods Human islets were isolated from living patients without diabetes. PERKi GSK2606414 effects were evaluated in the islets under glucolipotoxicity by palmitate. Islet insulin contents and secretion were measured. Autophagic flux was assessed by microtubule associated protein 1 light chain 3 (LC3) conversion, a red fluorescent protein (RFP)-green fluorescent protein (GFP)- LC3 tandem assay, and P62 levels. For mechanical analyses, autophagy was suppressed using 3-methyladenine in mouse islets. Small interfering RNA for an autophagy-related gene autophagy related 7 (Atg7) was transfected to interfere autophagy. Results PERKi administration to mice decreased diabetes-induced P62 levels in the islets. Glucolipotoxicity significantly increased PERK phosphorylation by 70% and decreased insulin contents by 50% in human islets, and addition of PERKi (40 to 80 nM) recovered both. PERKi also enhanced glucose-stimulated insulin secretion (6-fold). PERKi up-regulated LC3 conversion suppressed by glucolipotoxicity, and down-regulated P62 contents without changes in P62 transcription, indicating enhanced autophagic flux. Increased autophagosome-lysosome fusion by PERKi was visualized in mouse islets, where PERKi enhanced Atg7 bound to LC3. Suppression of Atg7 eliminated PERKi-induced insulin contents and secretion. Conclusion This study provided functional changes of human islets with regard to autophagy under glucolipotoxicity, and suggested modulation of autophagy as an anti-diabetic mechanism of PERKi. |
| first_indexed | 2024-04-24T13:08:46Z |
| format | Article |
| id | doaj.art-b7f06abe4ff54d33b25d401136d3d308 |
| institution | Directory Open Access Journal |
| issn | 2233-6079 2233-6087 |
| language | English |
| last_indexed | 2024-04-24T13:08:46Z |
| publishDate | 2024-03-01 |
| publisher | Korean Diabetes Association |
| record_format | Article |
| series | Diabetes & Metabolism Journal |
| spelling | doaj.art-b7f06abe4ff54d33b25d401136d3d3082024-04-05T06:03:30ZengKorean Diabetes AssociationDiabetes & Metabolism Journal2233-60792233-60872024-03-0148223124110.4093/dmj.2022.03662770Glucolipotoxicity Suppressed Autophagy and Insulin Contents in Human Islets, and Attenuation of PERK Activity Enhanced Them in an ATG7-Dependent MannerSeoil Moon0Ji Yoon Lim1Mirang Lee2Youngmin Han3Hongbeom Kim4Wooil Kwon5Jin-Young Jang6Mi Na Kim7Kyong Soo Park8Hye Seung Jung9 Department of Internal Medicine, Seoul National University Hospital, Seoul, Korea Department of Internal Medicine, Seoul National University Hospital, Seoul, Korea Department of Surgery and Cancer Research Institute, Seoul National University Hospital, Seoul, Korea Department of Surgery and Cancer Research Institute, Seoul National University Hospital, Seoul, Korea Department of Surgery and Cancer Research Institute, Seoul National University Hospital, Seoul, Korea Department of Surgery and Cancer Research Institute, Seoul National University Hospital, Seoul, Korea Department of Surgery and Cancer Research Institute, Seoul National University Hospital, Seoul, Korea Department of Internal Medicine, Seoul National University Hospital, Seoul, Korea Department of Internal Medicine, Seoul National University Hospital, Seoul, Korea Department of Internal Medicine, Seoul National University Hospital, Seoul, KoreaBackground Administration of pancreatic endoplasmic reticulum kinase inhibitor (PERKi) improved insulin secretion and hyperglycemia in obese diabetic mice. In this study, autophagic balance was studied whether to mediate it. Methods Human islets were isolated from living patients without diabetes. PERKi GSK2606414 effects were evaluated in the islets under glucolipotoxicity by palmitate. Islet insulin contents and secretion were measured. Autophagic flux was assessed by microtubule associated protein 1 light chain 3 (LC3) conversion, a red fluorescent protein (RFP)-green fluorescent protein (GFP)- LC3 tandem assay, and P62 levels. For mechanical analyses, autophagy was suppressed using 3-methyladenine in mouse islets. Small interfering RNA for an autophagy-related gene autophagy related 7 (Atg7) was transfected to interfere autophagy. Results PERKi administration to mice decreased diabetes-induced P62 levels in the islets. Glucolipotoxicity significantly increased PERK phosphorylation by 70% and decreased insulin contents by 50% in human islets, and addition of PERKi (40 to 80 nM) recovered both. PERKi also enhanced glucose-stimulated insulin secretion (6-fold). PERKi up-regulated LC3 conversion suppressed by glucolipotoxicity, and down-regulated P62 contents without changes in P62 transcription, indicating enhanced autophagic flux. Increased autophagosome-lysosome fusion by PERKi was visualized in mouse islets, where PERKi enhanced Atg7 bound to LC3. Suppression of Atg7 eliminated PERKi-induced insulin contents and secretion. Conclusion This study provided functional changes of human islets with regard to autophagy under glucolipotoxicity, and suggested modulation of autophagy as an anti-diabetic mechanism of PERKi.http://e-dmj.org/upload/pdf/dmj-2022-0366.pdfautophagydiabetes mellitushumansinsulinislets of langerhans |
| spellingShingle | Seoil Moon Ji Yoon Lim Mirang Lee Youngmin Han Hongbeom Kim Wooil Kwon Jin-Young Jang Mi Na Kim Kyong Soo Park Hye Seung Jung Glucolipotoxicity Suppressed Autophagy and Insulin Contents in Human Islets, and Attenuation of PERK Activity Enhanced Them in an ATG7-Dependent Manner Diabetes & Metabolism Journal autophagy diabetes mellitus humans insulin islets of langerhans |
| title | Glucolipotoxicity Suppressed Autophagy and Insulin Contents in Human Islets, and Attenuation of PERK Activity Enhanced Them in an ATG7-Dependent Manner |
| title_full | Glucolipotoxicity Suppressed Autophagy and Insulin Contents in Human Islets, and Attenuation of PERK Activity Enhanced Them in an ATG7-Dependent Manner |
| title_fullStr | Glucolipotoxicity Suppressed Autophagy and Insulin Contents in Human Islets, and Attenuation of PERK Activity Enhanced Them in an ATG7-Dependent Manner |
| title_full_unstemmed | Glucolipotoxicity Suppressed Autophagy and Insulin Contents in Human Islets, and Attenuation of PERK Activity Enhanced Them in an ATG7-Dependent Manner |
| title_short | Glucolipotoxicity Suppressed Autophagy and Insulin Contents in Human Islets, and Attenuation of PERK Activity Enhanced Them in an ATG7-Dependent Manner |
| title_sort | glucolipotoxicity suppressed autophagy and insulin contents in human islets and attenuation of perk activity enhanced them in an atg7 dependent manner |
| topic | autophagy diabetes mellitus humans insulin islets of langerhans |
| url | http://e-dmj.org/upload/pdf/dmj-2022-0366.pdf |
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