Impact of glucotoxicity induced in vivo and in vitro in Psammomys obesus

Berdja Sihem, Smail Leila, Othmani Kheira, Neggazi Samia, Hamlat Nadjiba, Boumaza Saliha, Sahraoui Abdelhamid, Kacimi Ghouti, Haffaf El Mahdi, Benazzoug Yasmina, Aouichat Bouguerra Souhila

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Laboratory of Cellular and Molécular Physiopathology, Institute of Natural Sciences, University of Technological Sciences, Houari Boumediene, Algiers, Algeria.

DOI: 10.4236/jdm.2012.21010                

Abstract

Objective: Chronic hyperglycemia characteristic of type diabetes 2 is responsible for the accelerated atherosclerosis with increased cardiovascular risk. In this study, we will propose to analyze the effect of a long-term of glucotoxicity in vivo in Psammomys obesus by addition of sucrose to 30% for 11 months and in vitro study of adventitial fibroblasts in the presence of D-glucose 0.6% for 7 days. Materials and methods: Evaluation of plasma biochemical parameters was carried out at the initial time and at the end of experiment. At autopsy, a morphological study of the aorta was performed after fixation in aqueous Bouin and staining with Masson’s trichrome. The experimental glucotoxicity is induced by incubation of fibroblasts in DMEM enriched with D-glucose at 0.6% for 7 days. The impact of glucotoxicity is assessed in the intracellular compartments through dosage of total nitrite and malondialdehyde, a product of lipid peroxidation, and thanks to a morphological assay after fixation of cells with aqueous bouin and blood staining with May Grünwald Giemsa. The evaluation of cell proliferation is accomplished by cell counting. Collagens I and III of the extracellular compartment are characterized by SDS-PAGE. Results: Animals subjected to sucrose showed hyperglycemia associated with hyperinsulinemia, dyslipidemia, hyperproteinemia, increased CPK and VLDL-LDL and decreased HDL. Histology of aortas revealed endothelial cells hypertrophy, severe disorganization of intima and media. In the presence of glucose, the proliferation of fibroblasts increases very significantly (P = 2.34 × 10^-5), the rate of malonaldehyde, nitrite and total density of chains α2 (I) and α1 (I + III) extra-cellular collagens I and III increased significantly. After staining, the cells showed hypertrophy, vacuolation of cytoplasm and chromatin condensation with nuclear fragmentation, indicative of apoptosis. Conclusion: The glucotoxicity induced in vivo and in vitro is responsible for major structural and metabolic alterations leading to the acceleration of the atherosclerotic process.

Keywords

Psammomys Obesus; Aorta; Adventitial Fibroblasts in Culture; DT2; Glucotoxicity; Extracellular Matrix; Oxidative Stress

Conflicts of Interest

The authors declare no conflicts of interest.

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