Effects of pomegranate seed oil on oxidant/antioxidant balance in heart and kidney homogenates and mitochondria of diabetic rats and high glucose-treated H9c2 cell line

Abstract

Objective: Oxidative stress is a major cause of diabetes complications. The present study aimed to investigate the beneficial effects of Pomegranate Seed Oil (PSO) on diabetes-induced changes in oxidant/antioxidant balance of the kidney, heart and mitochondria from rats and H9c2 cell line.

Materials and methods: In these in vivo and in vitro studies, male rats were divided into four groups (twelve each): group 1 served as control, group 2-4 received a single dose of streptozotocin (60 mg/kg, i.p), groups 3 and 4 received PSO (0.36 and 0.72 mg/kg/daily, gavage), respectively. After three weeks, six rats of each group and one week later the remaining animals were anaesthetized and the hearts and kidneys were removed and homogenized. Mitochondrial fractions were separated and enzyme activities were measured in each sample. H9c2 cells were pretreated with high levels of glucose (35 mM), and then, incubated with PSO. Finally, cell viability test, reactive oxygen species production and lipid peroxidation were evaluated.

Results: Significant reduction in enzymes activity (Superoxide dismutase, Glutathione S-transferase and Paraoxonase 1), compensatory elevation in Glutathione Reductase, Glutathione Peroxidase and Catalase activity followed by reduction after one week and significant elevation in Oxidative Stress Index (OSI) were observed in diabetic group. PSO treatment resulted in a significant increase in enzymes activity and decreased OSI values compared to diabetic group in both tissue and mitochondrial fractions. PSO remarkably decreased glucose-induced toxicity, ROS level and lipid peroxidation in H9c2 cells.

Conclusion: Results suggested that PSO has a protective effect against diabetes-induced alterations in oxidant/antioxidant balance in tissues, mitochondrial and H9c2 cell line.

Keywords: Antioxidant; Diabetes mellitus; Oxidant; Oxidative stress; Pomegranate Seed Oil.

Figures

Figure 1

Effect of the pomegranate seed oil treatment on tissue CAT activity in heart (left) and kidney (right) homogenates in a model of STZ-induced diabetes. Values are expressed as mean ± SEM. ** p

Figure 2

Effect of the pomegranate seed oil treatment on CAT activity in heart (left) and kidney (right) mitochondria in a model of STZ-induced diabetes. Values are expressed as mean ± SEM. ** p

Figure 3

Effect of the pomegranate seed oil treatment on tissue SOD activity in heart (left) and kidney (right) homogenates in a model of STZ-induced diabetes. Values are expressed as mean ± SEM. * p

Figure 4

Effect of the pomegranate seed oil treatment on SOD activity in heart (left) and kidney (right) mitochondria in a model of STZ-induced diabetes. Values are expressed as mean ± SEM. ** p

Figure 5

Effect of the pomegranate seed oil treatment on tissue GST activity in heart (left) and kidney (right) homogenates in a model of STZ-induced diabetes. Values are expressed as mean ± SEM. * p

Figure 6

Effect of the pomegranate seed oil treatment on GST activity in heart (left) and kidney (right) mitochondria in a model of STZ-induced diabetes. Values are expressed as mean ± SEM. * p

Figure 7

Effect of the pomegranate seed oil treatment on tissue PON1 activity in heart (left) and kidney (right) homogenates in a model of STZ-induced diabetes. Values are expressed as mean ± SEM. * p

Figure 8

Effect of the pomegranate seed oil treatment on PON1 activity in heart (left) and kidney (right) mitochondria in a model of STZ-induced diabetes. Values are expressed as mean ± SEM

Figure 9

Effect of PSO alone on cell viability in H9c2 (left). The cells were treated (for 24 hr) with different concentrations of PSO. Effect of PSO on glucose-induced cytotoxicity in H9c2 cells (right). The cells were pretreated (for 24 hr) with glucose (35 mM) before to exposure (for 24 and 48 hr) to non-toxic concentration of PSO. Data are expressed as mean ± SEM of three separate experiments. * p

Figure 10

Effect of PSO on glucose-induced ROS generation in H9c2 cells. The cells were pretreated (for 24 and 48 hr) with different concentrations of PSO, before exposure (24 hr) to glucose solution (35 mM). Data are expressed as mean ± SEM of three separate experiments. * p

Figure 11

Effect of PSO on glucose-induced MDA production in H9c2 cells. The cells were pretreated (for 24 hr) with different concentrations of PSO before exposure (for 3 hr) to glucose solution (35 mM). Data are expressed as mean ± SEM of three independent experiments. * p