The reduction of hypoxia-induced and reoxygenation-induced apoptosis in rat islets by epigallocatechin gallate.
The survival of transplanted tissue is affected by the detrimental consequences of hypoxia followed by reoxygenation. The majority of transplanted cells undergo apoptosis due to hypoxia and reoxygenation (H/R) injury, but protection from H/R has been less examined. In this study, we examined whether epigallocatechin gallate (EGCG) protected rat islets from H/R injury. Rat islets, freshly prepared from F344 rat strain by collagenase digestion and density centrifugation, were seeded in triplicate at concentrations of 100 per well in 24-well plates for culture under normoxia. The cells were then exposed to hypoxia for 14 hours with or without EGCG, after which they were reoxygenated for 72 hours in a humidified oxygenated CO(2) incubator at 37 degrees C. Apoptosis, lactate dehydrogenase (LDH), and 8-hydroxy-2'-deoxyguanosine (8-OHdG) were evaluated according to the manufacturer's instructions. The H/R induced apoptosis in the islets that was reduced in dose-dependent manner by EGCG treatment. The viability of islets exposed to H/R was assessed by LDH release. H/R reduced viability compared with the controls, while the viability of the islets improved upon EGCG treatment. The secretion of insulin was also decreased by H/R, as well as the dose dependent EGCG protective ability on insulin secretion. The content of 8-OHdG in islets from H/R was also reduced by EGCG. Our results indicated that apoptosis and the decline in insulin secretion by H/R were inhibited by EGCG treatment. EGCG may be considered useful for protection of islets from oxidative injury associated with the transplantation procedure.
Hara
Y
Y
Laboratory of Transplantation Immunology and National Research Institute for Child Health and Development, Tokyo, Japan.
Fujino
M
M
Adachi
K
K
Li
X-K
XK
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002392:Catechin / Q000031:analogs & derivatives* / Q000494:pharmacology; D015687:Cell Hypoxia / Q000187:drug effects / Q000502:physiology; D007515:Islets of Langerhans / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D016381:Islets of Langerhans Transplantation / Q000473:pathology / Q000502:physiology*; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D010101:Oxygen Consumption / Q000187:drug effects / Q000502:physiology; D051381:Rats; D011916:Rats, Inbred F344
D002392:Catechin; C045651:epigallocatechin gallate; D007770:L-Lactate Dehydrogenase