Anhydrous microwave synthesis as efficient method for obtaining model advanced glycation end-products
IntroductionAdvanced glycation end-products (AGEs) are capable of stimulating oxidative stress and inflammation. This study investigates the synthesis of medium crosslinked AGEs (the most optimal form of AGEs because of soluble in water, used in many assays as markers) and their biochemical properti...
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Frontiers Media S.A.
2024-11-01
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| Series: | Frontiers in Molecular Biosciences |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmolb.2024.1484196/full |
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| author | Agnieszka Bronowicka-Szydełko Katarzyna Madziarska Aleksandra Kuzan Łukasz Lewandowski Joanna Adamiec-Mroczek Jadwiga Pietkiewicz Maciej Tota Maciej Ziomek Wojciech Stach Tymoteusz Trocha Marcin Piersiak Maciej Pachana Zuzanna Galińska Andrzej Korpacki Olgierd Dróżdż Janusz Matuszyk Małgorzata Mitkiewicz Andrzej Gamian Kinga Gostomska-Pampuch |
| author_facet | Agnieszka Bronowicka-Szydełko Katarzyna Madziarska Aleksandra Kuzan Łukasz Lewandowski Joanna Adamiec-Mroczek Jadwiga Pietkiewicz Maciej Tota Maciej Ziomek Wojciech Stach Tymoteusz Trocha Marcin Piersiak Maciej Pachana Zuzanna Galińska Andrzej Korpacki Olgierd Dróżdż Janusz Matuszyk Małgorzata Mitkiewicz Andrzej Gamian Kinga Gostomska-Pampuch |
| author_sort | Agnieszka Bronowicka-Szydełko |
| collection | DOAJ |
| description | IntroductionAdvanced glycation end-products (AGEs) are capable of stimulating oxidative stress and inflammation. This study investigates the synthesis of medium crosslinked AGEs (the most optimal form of AGEs because of soluble in water, used in many assays as markers) and their biochemical properties.MethodsOne of model protein–myoglobin from horse heart muscle (MB) and a chosen respective glycation factor – D-melibiose (mel), acrolein (ACR), D-glucose (glc), 4-hydroksynonenal (4HNE), trans-2-nonenal (T2N), methylglyoxal (MGO) – were subjected to high temperature water synthesis (HTWS) and high temperature microwave synthesis in anhydrous conditions (HTMS). The syntheses were deliberately carried out in two different conditions to check whether adding an additional energy source (microwaves) while lowering the temperature and shortening the reaction time would allow for more effective obtaining of medium-cross-linked AGEs, monitored with SDS-PAGE. Products were analyzed using fluorescence measurements, Enzyme-Linked Immunosorbent Assay (ELISA) and immunoblotting tests and electrophoretic mobility shift assay to evaluate their ability to activate nuclear factor kappa-light-chain-enhancer (NF-κB).ResultsMedium cross-linked AGEs were more efficiently obtained in HTMS. Fluorescence was high for MB-ACR, MB-T2N and MB-glc products. Anti-MAGE antibodies showed reactivity towards MB-mels of HTMS and HTWS, and the MB-4HNEs from HTMS. HTWS products, apart from MB-ACR, did not activate NF-κB, whereas MB-ACR, MB-4HNE, MB-mel, and MB-T2N products of HTMS strongly activated this factor that indicates their strong pro-inflammatory properties.ConclusionHTMS is a fast and efficient method of synthesizing medium cross-linked AGEs. |
| format | Article |
| id | doaj-art-70bd9769ed434e678fb8d6e72dc1790f |
| institution | OA Journals |
| issn | 2296-889X |
| language | English |
| publishDate | 2024-11-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Molecular Biosciences |
| spelling | doaj-art-70bd9769ed434e678fb8d6e72dc1790f2025-08-20T02:14:39ZengFrontiers Media S.A.Frontiers in Molecular Biosciences2296-889X2024-11-011110.3389/fmolb.2024.14841961484196Anhydrous microwave synthesis as efficient method for obtaining model advanced glycation end-productsAgnieszka Bronowicka-Szydełko0Katarzyna Madziarska1Aleksandra Kuzan2Łukasz Lewandowski3Joanna Adamiec-Mroczek4Jadwiga Pietkiewicz5Maciej Tota6Maciej Ziomek7Wojciech Stach8Tymoteusz Trocha9Marcin Piersiak10Maciej Pachana11Zuzanna Galińska12Andrzej Korpacki13Olgierd Dróżdż14Janusz Matuszyk15Małgorzata Mitkiewicz16Andrzej Gamian17Kinga Gostomska-Pampuch18Department of Biochemistry and Immunochemistry, Wroclaw Medical University, Wroclaw, PolandClinical Department of Diabetology, Hypertension and Internal Disease, Wroclaw Medical University, Wroclaw, PolandDepartment of Preclinical Sciences, Pharmacology and Medical Diagnostics, Faculty of Medicine, Wroclaw University of Science and Technology, Wroclaw, PolandDepartment of Biochemistry and Immunochemistry, Wroclaw Medical University, Wroclaw, PolandClinical Department of Ophthalmology, Wroclaw Medical University, Wroclaw, PolandDepartment of Biochemistry and Immunochemistry, Wroclaw Medical University, Wroclaw, PolandFaculty of Medicine, Wroclaw Medical University, Wroclaw, PolandFaculty of Medicine, Wroclaw Medical University, Wroclaw, PolandFaculty of Medicine, Wroclaw Medical University, Wroclaw, PolandFaculty of Medicine, Wroclaw Medical University, Wroclaw, PolandFaculty of Medicine, Wroclaw Medical University, Wroclaw, PolandFaculty of Medicine, Wroclaw Medical University, Wroclaw, PolandFaculty of Medicine, Wroclaw Medical University, Wroclaw, PolandFaculty of Medicine, Wroclaw Medical University, Wroclaw, PolandClinical Department of Ophthalmology, Wroclaw Medical University, Wroclaw, PolandLaboratory of Tumor Molecular Immunobiology, Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wroclaw, PolandLaboratory of Tumor Molecular Immunobiology, Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wroclaw, PolandLaboratory of Medical Microbiology, Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wroclaw, PolandDepartment of Biochemistry and Immunochemistry, Wroclaw Medical University, Wroclaw, PolandIntroductionAdvanced glycation end-products (AGEs) are capable of stimulating oxidative stress and inflammation. This study investigates the synthesis of medium crosslinked AGEs (the most optimal form of AGEs because of soluble in water, used in many assays as markers) and their biochemical properties.MethodsOne of model protein–myoglobin from horse heart muscle (MB) and a chosen respective glycation factor – D-melibiose (mel), acrolein (ACR), D-glucose (glc), 4-hydroksynonenal (4HNE), trans-2-nonenal (T2N), methylglyoxal (MGO) – were subjected to high temperature water synthesis (HTWS) and high temperature microwave synthesis in anhydrous conditions (HTMS). The syntheses were deliberately carried out in two different conditions to check whether adding an additional energy source (microwaves) while lowering the temperature and shortening the reaction time would allow for more effective obtaining of medium-cross-linked AGEs, monitored with SDS-PAGE. Products were analyzed using fluorescence measurements, Enzyme-Linked Immunosorbent Assay (ELISA) and immunoblotting tests and electrophoretic mobility shift assay to evaluate their ability to activate nuclear factor kappa-light-chain-enhancer (NF-κB).ResultsMedium cross-linked AGEs were more efficiently obtained in HTMS. Fluorescence was high for MB-ACR, MB-T2N and MB-glc products. Anti-MAGE antibodies showed reactivity towards MB-mels of HTMS and HTWS, and the MB-4HNEs from HTMS. HTWS products, apart from MB-ACR, did not activate NF-κB, whereas MB-ACR, MB-4HNE, MB-mel, and MB-T2N products of HTMS strongly activated this factor that indicates their strong pro-inflammatory properties.ConclusionHTMS is a fast and efficient method of synthesizing medium cross-linked AGEs.https://www.frontiersin.org/articles/10.3389/fmolb.2024.1484196/fulladvanced glycation end-productsNF-κBmicrowave synthesispro-inflammatory factorsAGEsbiomarkers |
| spellingShingle | Agnieszka Bronowicka-Szydełko Katarzyna Madziarska Aleksandra Kuzan Łukasz Lewandowski Joanna Adamiec-Mroczek Jadwiga Pietkiewicz Maciej Tota Maciej Ziomek Wojciech Stach Tymoteusz Trocha Marcin Piersiak Maciej Pachana Zuzanna Galińska Andrzej Korpacki Olgierd Dróżdż Janusz Matuszyk Małgorzata Mitkiewicz Andrzej Gamian Kinga Gostomska-Pampuch Anhydrous microwave synthesis as efficient method for obtaining model advanced glycation end-products Frontiers in Molecular Biosciences advanced glycation end-products NF-κB microwave synthesis pro-inflammatory factors AGEs biomarkers |
| title | Anhydrous microwave synthesis as efficient method for obtaining model advanced glycation end-products |
| title_full | Anhydrous microwave synthesis as efficient method for obtaining model advanced glycation end-products |
| title_fullStr | Anhydrous microwave synthesis as efficient method for obtaining model advanced glycation end-products |
| title_full_unstemmed | Anhydrous microwave synthesis as efficient method for obtaining model advanced glycation end-products |
| title_short | Anhydrous microwave synthesis as efficient method for obtaining model advanced glycation end-products |
| title_sort | anhydrous microwave synthesis as efficient method for obtaining model advanced glycation end products |
| topic | advanced glycation end-products NF-κB microwave synthesis pro-inflammatory factors AGEs biomarkers |
| url | https://www.frontiersin.org/articles/10.3389/fmolb.2024.1484196/full |
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