Protective Effects of a Calcium Channel Blocker on Apoptosis in Thymus of Neonatal Stz-Diabetic Rats

Abstract

Streptozotocin (STZ) is known to induce insulin-dependent diabetes in experimental animals. In STZ-induced diabetes atrophy of the thymus is caused by elevated intracellular calcium levels leading to apoptosis. Hyperglycemia is known to result in a decrease in numbers of T cells in the thymus and circulation. Intracellular calcium levels increase in diabetic animals after induction by STZ. Hyperglycemia inhibits Ca2+-ATPase and increases intracellular calcium levels. We have investigated apoptosis in thymus tissue of neonatal STZ (n-STZ)-diabetic rats and the effects of isradipine as a calcium channel blocker (CCB) on apoptosis. Five groups of newborn Wistar rats were used. On the second day after birth 100mg/kg STZ was given i.p. to the first two groups. The first group was n-STZ diabetic. To the second group starting from the 12th week 5 mg/kg/day isradipine (i.p) was given for 6 weeks. To the third group the same dose of isradipine was given on the second day followed by STZ treatment. The fourth group was non-diabetic and treated with 5 mg/kg/day isradipine for six weeks. The fifth group consisted of non-diabetic rats. To the sixth group dexamethasone (5 mg/kg i.p.) was given to adult rats. For detection of apoptotic cells in paraffin-embedded thymus sections the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end Labelling (TUNEL) assay was used. The DNA ladder method was performed for anaLysis of DNA fragmentation. In the isradipine-treated non-diabetic group typical apoptotic banding patterns were found whereas thick bands between 123 and 246 bp length were found in the n-STZ- and n-STZ+isradipine-treated groups. More apoptotic cells were observed in the thymus of isradipine-treated n-STZ-treated and n-STZ+isradipine-treated groups when compared with the non-diabetic control and isradipine+n-STZ-treated groups. In conclusion we observed that long-term STZ diabetes results in apoptosis in the thymus. We also found that isradipine administered before STZ has protective effects against apoptosis whereas isradipine alone induces apoptosis. (c) 2005 Elsevier GmbH. All rights reserved.