Analysis of morphometric and ultrastructural characteristics of blood microcirculation in hippocampus of rats with alloxan diabetes under administration of citicoline

Abstract

Aim: determine morphometric and ultrastructural features of blood microcirculation in hippocampus of rats with alloxan diabetes under experimental therapy with citicoline. Materials and methods . The research was carried out on 48 white male Wistar rats (250-300 g) randomized in 3 groups by 16 animals: I – intact (distilled water, intragastrically); II – animals with diabetes (distilled water, intragastrically); III – animals with diabetes + citicoline (500 mg/kg, intragastrically). Diabetes was reproduced by single subcutaneous injection of alloxan monohydrate (150 mg/kg). Blood glucose level was determined on the 11 th day after administration of alloxan using a glucometer. Citicoline and distilled water were introduced once a day during 20 days starting from the 11 th day after administration of alloxan. In morpho-functional study of hippocampal CA1-zone neurons the histological sections were deparaffinized and dyed with gallocyanin-chrome alum by Einarson for specific detection of RNA. Pictures were received with a microscope Axioskop (Zeiss, Germany). Using a computer-based picture analysis system VIDAS-386 (Kontron Elektronik, Germany) the density of endothelial nuclei, the nucleus area and the concentration of RNA in the nucleus were determined. The study of ultrastructural changes of blood microcirculation in hippocampus was carried out using transmission electron microscope PEM-100-01 («SELMI», Ukraine) by standard procedure and in accordance with common standards. Statistical difference between the groups were assessed by non-parametric Mann-Whitney U-test. Results . It has been established that citicoline contributed to recovery of proliferative activity of endothelium in vascular bed of hippocampus manifested with increase of endothelial nuclei density, their area and levels of nuclear RNA by 19,4% (p < 0,001), 17,2% (p < 0,05) and 25,8% (p < 0,01). Simultaneously it has been determined that hematoencephalic barrier damage in hippocampus of rats under administration of citicoline were less expressed or virtually absent. Conclusion . Thus, stimulation of revascularization processes, optimization of metabolic processes and also lack of pronounced changes in microcirculatory bed in hippocampus of rats with alloxan diabetes are the evidence of expressed endothelium-protective activity of citicoline