FEATURES OF THE CELL CYCLE OF LIVER CELLS AFTER PARTIAL HEPATECTOMY

Pivtorak V. I., Bulko M. P., Kostyuk G. Ya.

MORPHOLOGY

Scentific article

Introduction. The liver resection, in most cases, remains the method of choice and the only way to achieve radical treatment and increase the life expectancy of patients. The human liver has significant regenerative capacity and is restored, even after removing 70% of its mass. However, surgeries on the liver, even a small amount can lead to serious consequences, including, and because a number of factors natural resistance is synthesized in the liver. The purpose of the study is to determine the characteristics of the cellular cycle of liver cells remaining after partial resection in sexually mature rats. Object and methods. The experiment was conducted on 42 male rats weighing 200-250 g. Animals were kept under vivarium conditions with free access to food and water in a diet that meets the standards for rats. In the control group, no rats were given any of the six rats. All animals of the experimental group (36 rats) performed surgical intervention for liver resection. The middle front lobe of the liver and the left lobe of the liver were removed (~ 70% of the total weight of the liver). Animals were taken out from the experiment of 6 animals for each term: after 1, 2, 3, 7, 10, 14 days after resection of the liver by intra-pleural administration of thiopental-sodium (50 mg/kg). Maintenance and manipulation of the animals were carried out according to the “General ethical animal experimentation” adopted the first National Congress on Bioethics (Kyiv, 2001), also guided by the recommendations of the “European Convention for the Protection of vertebrate animals used for experimental and other scientific purposes” (Strasbourg, 1985) and the provisions of the “Rules of preclinical safety evaluation of pharmacological agents (GLP)”. The DNA content of the rat liver cell nuclei was determined by flow DNA cytometry. Liver cell nucleus suspensions from rats were prepared using a CyStain DNA Step 1 (Partec, Germany) nuclear DNA dilution solution in accordance with the manufacturer’s protocol. This solution allows for the extraction of nuclei and the labeling of nuclear DNA by diaminophenylindole (DAPI). In the process of making nuclear suspensions disposable filters CellTrics 50 microns (Partec, Germany) were used. Flow analysis was performed on multifunctional research flow cytometer “Partec PAS” (Partec, Germany). Results and discussion. Analysis of cell cycle and fragmentation of liver cell DNA showed a certain balance of processes of synthesis and fragmentation of nuclear DNA in liver cells of intact animals. There is a predominance of the proportion of cells that are in the phase of proliferative rest (G0G1). In the state of proliferative activity (S phase, G2+M phase) there is a significantly smaller number of cells. Characterizing the synthetic activity of liver cells, it is necessary to note statistically significantly more cells in the phase S in one (2,9 times) and two days (3,8 times) after liver resection compared with animals in the control group. Subsequently, three days after the resection, the number of cells was smaller (2,4 times; p<0,05) compared with the animals withdrawn from the study for the second day, but 1,6 times (p<0,05) higher in comparison with control animals. After seven days, the number of cells in this phase was greater than 1,3 times (p <0,05) compared to the third day. Ten days later, compared with the seventh day, a smaller number of cells was detected in 2,7 times (p <0,05). In fourteen days compared to the tenth day, the number of cells was 1,3 times larger (p<0,05) and was closer to the indicator in the control group of animals. Conclusions. The features of the cell cycle of liver cells in the sexually mature animals in the immediate postoperative period after liver resection are determined: the number of cells in the synthetic cycle of the cell cycle (phase S) grows wavelike. The peak of growth was observed for 1-2 days, the second wave of growth – for 7 days, the third – at 14 days postoperative period. At 10-14 days of observation after liver resection, it was found that the number of cells in the G0G1 phase is statistically significantly lower, and in the G2+M phase it is statistically significantly more significant compared to those of the earlier postoperative period.

liver, hepatectomy, regeneration, cell cycle

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«Bulletin of problems biology and medicine» Issue 4 Part 2 (147), 2018 year, 287-290 pages, index UDK 616.36-03.93:617-089:576.36

10.29254/2077-4214-2018-4-2-147-287-290