Morphometric Properties of Cells of the Line Hepg2 Under the Action of Plumbum Sulfate

Abstract

Lead is a typical diffuse element contained in all components of the environment: in rocks, soils, natural waters, atmosphere, and living organisms. All lead compounds act practically the same, the difference in toxicity is explained by their different solubility in body fluids. In particular, lead sulfate has the most harmful effect on liver cells. In most studies using cell lines of hepatocyte origin, including the HepG2 cell line, metabolic parameters are investigated. Also, the study of the morphology of hepatocytes (morphometric indicators) is no less important in the study of the action of the toxicant.Therefore, the purpose of the study was to analyze the morphofunctional properties of cultured hepatocytes under the action of lead sulfate. In the course of this work, the main morphometric parameters (nuclear-cytoplasmic ratio, cell area, nuclear area) and the total adhesive potential were measured and determined in the control and with the addition of lead sulfate at a concentration of IC50/10 (0.004 mM). Cell counting was performed using Axiovision and ImageJ programs. The nuclear-cytoplasmic ratio is calculated – NC ratio =N/C, where N is the area of the nucleus, С - the area of the cytoplasm. The adhesive potential was determined by the ability of cells to attach to the plastic surface under the action of lead sulfate. By calculating the ratio of live and dead cells after staining with trypan blue, the cytotoxic effect of lead sulfate was recorded, which consisted in the death of cells in the toxicant range of 0.016-1 mM (30-80% of the control) with an inhibitory concentration indicator (IC50) equal to 0.04 mM. Using a colorimetric research method, it was shown that the total adhesive index of cells incubated with lead sulfate decreased by 1.5-3 times (p<0.05) compared to the control. The content of cells with an NC ratio of 0.4-0.5 under the influence of lead sulfate doubled compared to the control; cells with defective appendages and a polygonal cell shape, which distinguishes them from the control, were also recorded.