OBJECTIVE: Free radicals that escape scavenging by antioxidant defense damage lipids, proteins, and DNA. Damage to DNA can be repaired. Therefore, both cells' antioxidant defense and their ability to repair oxidatively damaged DNA decide its fate to survive oxidative stress. Pancreatic islets cells with poor antioxidant defense were checked for their ability to remove oxidative damage form DNA. METHODS: For ex vivo DNA repair, assay-cultured pancreatic islets and liver slices were treated with 1 and 10 mM H2O2, respectively, for 30 minutes. After incubation for different time intervals, 8-hydroxy-2'-deoxyguanosine (8-OHdG) in DNA of these cells was estimated using monoclonal antibody raised against 8-OHdG by competitive enzyme-linked immunosorbent assay. For in vitro DNA repair assay, oxidatively damaged pBR322 was incubated with nuclear extracts of islet and liver cells, and 8-OHdG retained in the plasmid was quantitated. RESULTS: Oxidative damage induced by H2O2 was removed quickly and efficiently from DNA by liver cells compared with islet cells. The repair of oxidatively damaged plasmid DNA in vitro was also performed more efficiently (P < 0.05) by nuclear extracts from liver cells compared with islet cell. CONCLUSIONS: We clearly demonstrate that in addition to their low antioxidant defense, islets are very poor in rectifying the oxidative DNA damage.
OBJECTIVE:Free radicals that escape scavenging by antioxidant defense damage lipids, proteins, and DNA. Damage to DNA can be repaired. Therefore, both cells' antioxidant defense and their ability to repair oxidatively damaged DNA decide its fate to survive oxidative stress. Pancreatic islets cells with poor antioxidant defense were checked for their ability to remove oxidative damage form DNA. METHODS: For ex vivo DNA repair, assay-cultured pancreatic islets and liver slices were treated with 1 and 10 mM H2O2, respectively, for 30 minutes. After incubation for different time intervals, 8-hydroxy-2'-deoxyguanosine (8-OHdG) in DNA of these cells was estimated using monoclonal antibody raised against 8-OHdG by competitive enzyme-linked immunosorbent assay. For in vitro DNA repair assay, oxidatively damaged pBR322 was incubated with nuclear extracts of islet and liver cells, and 8-OHdG retained in the plasmid was quantitated. RESULTS: Oxidative damage induced by H2O2 was removed quickly and efficiently from DNA by liver cells compared with islet cells. The repair of oxidatively damaged plasmid DNA in vitro was also performed more efficiently (P < 0.05) by nuclear extracts from liver cells compared with islet cell. CONCLUSIONS: We clearly demonstrate that in addition to their low antioxidant defense, islets are very poor in rectifying the oxidative DNA damage.
Authors: Y Imai; A D Dobrian; J R Weaver; M J Butcher; B K Cole; E V Galkina; M A Morris; D A Taylor-Fishwick; J L Nadler Journal: Diabetes Obes Metab Date: 2013-09 Impact factor: 6.577
Authors: D A Taylor-Fishwick; J R Weaver; W Grzesik; S Chakrabarti; S Green-Mitchell; Y Imai; N Kuhn; J L Nadler Journal: Diabetologia Date: 2012-10-03 Impact factor: 10.122
Authors: John P McQuilling; Sivanandane Sittadjody; Samuel Pendergraft; Alan C Farney; Emmanuel C Opara Journal: Biomater Sci Date: 2017-11-21 Impact factor: 6.843