Histological and immunohistochemical study on the possible ameliorating effects of antox on the parotid gland of rats with Streptozotocin- Induced Diabetes Mellitus Type 1

Abstract

Background: Hyperglycemia associated with the diabetes mellitus (DM) commonly results in structural abnormalities and hyposalivation in the parotid glands. An antioxidant action in a potent multivitamin called Antox (ANX) aids in reducing oxidative stress. Aim of study: The current study examined the role of ANX in preventing complications from diabetes in the parotid gland, as well as its potential mechanisms. Material and methods: 36 rats were randomly divided into six groups; each group contained six rats. Group 1 (Negative (-ve) control): the rats received only regular food and water. Group 2 (vehicle group): rats received a single i.p. dose of citrate buffer as a vehicle. Group 3: rats were given ANX by oral gavage with a polyethylene canula 0.5 mm in a volume not to exceed 0.3 ml/100 gm at a dose of 10 mg/kg/day. Group 4 (Diabetic/ DM): rats received a single i.p. dose of 50 mg/kg of STZ dissolved in citrate buffer. Group 5 (DM+ Insulin): rats received a single i.p. dose of 50 mg/kg of STZ dissolved in citrate buffer and insulin (Mixtard 30/70; Novo Nordisk) 1 U/100 gm once daily subcutaneous (S.C.). Group 6 (DM+ Insulin + ANX): rats received a single i.p. dose of 50 mg/kg of STZ and insulin 1 U/100 gm /day/S.C. then received ANX orally in a dose of 10 mg/kg/day. All medications were given for 4 weeks. Rats were anaesthetized, and the parotid tissues were obtained for biochemical analysis to measure Malondialdehyde (MDA), superoxide dismutase (SOD), reduced glutathione (GSH), and nitric oxide (NO), for Histopathological examination (Hematoxylin and Eosin staining, Masson trichrome stain) and immunohistochemistry using (8-OHdG, α-SMA and BAX). Results: Remarkably, co-administration of ANX with insulin significantly reduced malondialdehyde (MDA) levels and increased expression of 8-hydroxy-2'-deoxyguanosine (8-OHdG) while enhancing the activities of superoxide dismutase (SOD), glutathione peroxidase (GPX), and total antioxidant capacity (TAC). Additionally, compared to the diabetic rat group and the diabetic group receiving insulin, the combination of ANX and insulin resulted in a considerably decreased expression of Bcl-2, Associated X-protein (Bax) and Smooth muscle alpha-actin (α-SMA), with significant reduction in interleukin-1beta (IL1β) level in relation to diabetic group. Conclusion: In comparison to insulin administration alone, co-delivery of ANX with insulin throughout diabetes treatment was more effective in preserving the structure and function of the parotid gland.