STRUCTURAL ORGANIZATION AND QUANTITATIVE ANALYSIS OF MAIN CELLS OF HEPATIC SINUSOIDAL WALL OF RATS AFTER 8 WEEKS OF EXPERIMENTAL EXPOSURE TO CANNABIDIOL OIL

Abstract

Introduction. Cannabidiol (CBD) is a non-psychotropic phytocannabinoid of Cannabis sativa with therapeutic potential, which is used to treat children with severe forms of epilepsy in Dravet and Lennox-Gastaut syndrome. Preclinical trials and clinical studies demonstrate the positive effects of CBD on the body. However, there are reports of possible side effects from various organs and systems, especially at high doses or after prolonged use. Of particular concern is the risk of CBD-induced hepatotoxicity.Experimental morphological studies of the liver are necessary, especially the study of the effect of CBD on the liver sinusoids to establish the safety of use.The purpose of the study: to determine the structural organization and conduct a quantitative analysis of the cellular composition of rat liver sinusoidal capillaries after 8 weeks of experimental exposure to 10% CBD oil.Methodology. The experimental series was conducted in vivarium conditions on 20 white non-linear male rats weighing 180–230 g, aged 5–7 months in accordance with the provisions of the European Convention for the Protection of Vertebrate Animals used for Experimental and Other Scientific Purposes. The main group was 14 rats that received 10% CBD oil orally once a day (dose 10 mg/kg) for 8 weeks. The manufacturer of the studied product was the licensed Aroma Extract Labs s.r.o. (Praha, Czech Republic). The control group was 6 intact sexually mature white male rats. At the end of the experiment, after euthanasia, liver tissue was collected for morphological study. Histological, immunohistochemical and morphometric studies of the cellular composition of the liver sinusoids were performed. For quantitative analysis of liver sinusoidal capillaries, the main cells of the sinusoidal wall were counted in different zones of hepatic lobules on an area of 0.01 mm2 of histological preparation (100 μm×100 μm). The data are presented as mean and percentage values and their errors. The significance of the difference in mean values was determined by the Mann-Whitney test, and percentage values by the Pearson test (χ2). In all comparisons, the difference was considered statistically significant at a minimum significance level of p<0.05.Scientific novelty: The absence of dystrophic changes in sinusoidal capillaries was established, morphological signs of hemostasis disorders and hemodynamic disorders were not diagnosed, as well as destructive changes in endothelial cells and Kupffer cells. In all areas of the hepatic lobule, the average values of Kupffer cells significantly exceeded the corresponding values of the control group p (U) <0.0001, which can be explained by Kupffer cell hyperplasia and their key role in immune regulation. The average value of CD31 cells in the sinusoidal wall was significantly lower than the corresponding value of the control group p (U) <0.0001 in all areas of the hepatic lobule, due to an increase in the diameter of the sinusoids due to the vasodilatory effect of CBD.Perisinusoidal cells in the experimental series were the least numerous in all areas of the hepatic lobule compared to endothelial cells and Kupffer cells (p<0.05). However, studying the morphological features of perisinusoidal cells is important for assessing the experimental effect of CBD oil on the liver.Conclusions. The results of morphological and morphometric analysis of liver sinusoids after two and four weeks of experimental exposure to CBD indicate the safety of using a 10% CBD oil at a dose of 10 mg/kg/day.