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Using fluorescence methods for visualization of human tumor cell apoptosis and necrosis in vitro, the antitumor effects of flavonoid-containing Gratiola officinalis L. and green tea extracts at ultralow concentrations on A498 human kidney carcinoma cells after 24 and 48 h were compared. Fluorescent dyes (propidium iodide and acridine orange) were used in the “living and dead” test, which made it possible to identify the total number of cells that died by necrosis and apoptosis, as well as cells in which apoptosis has started and apoptotic bodies or pycnosis of the nucleus were detected. Gratiola officinalis L. extract had the most pronounced cytostatic activity 48 h after exposure; the number of cells per field of view decreased by a factor of 1.5 compared with the control. Green tea extract at a concentration of 0.0288 mg/mL had the most pronounced cytotoxic activity; its manifestation was growing 48 h after exposure. Gratiola officinalis L. extract exhibited cytotoxic activity at lower concentrations (0.0036 mg/mL).

The influence of long-term peroral injection of gold nanoparticles with various sizes on the liver, spleen, and lymph nodes of adult laboratory rats and their progeny was studied. The morphometric analysis was conducted using a µVizo-103 medical digital microviser. The conglomerated gold nanoparticles were detected in tissues via the dark-field microscopy. Gold nanoparticles with sizes of 2 and 50 nm were found to cause the mostly pronounced morphological alterations in organs that testifies to their size-dependent impact. The liver in a group receiving 50-nm gold particles exhibit noticeable dystrophy of hepatocytes and activation of a monocyte macrophage system. The spleen and lymph nodes reveal the activation of proliferation and differentiation of immunocompetent cells and macrophages. Peroral injection of gold particles during gestation of females led to an increase in a number of hepatocytes with necrobiotic changes as well as to elevated amount of non-parenchymatic elements in the liver of the progeny, especially in groups containing the 2- and 50-nm gold nanoparticles.

The possibilities of using fluorescence-based analytical methods and their advantages for visualization and identification of the type of programmed cell death in human tumor cells exposed to flavonoids have been analyzed in experiments in vitro. HeLa cervical cancer cells and A498 kidney carcinoma cells were used as the objects of the study, and exposure to the flavonoid-containing extract of common hedgehyssop (Gratiola officinalis L.) was tested as the experimental treatment. The following fluorescence-based techniques were used: the “live and dead” test with double staining by propidium iodide and acridine orange and double staining with annexin and propidium iodide. Autophagy induction was confirmed by fluorescence-based tests implemented in a Muse cell analyzer with a Muse Autophagy LC3-Antibody Based Kit. The use of double staining with acridine orange and propidium iodide fluorescent dyes in the “live and dead” test and comparison with phase contrast microscopy enables the visualization of apoptotic body and autophagosome formation processes in the cells and can, therefore, be used as a method of screening assessment of the efficiency of various chemotherapy drugs.

Cachexia is a severe complication of cancer and currently there are no drugs that would effectively deal with exhaustion and intoxication in various diseases. Materials and methods. In this paper a study and evaluation of the antitumor and anticachexia activities of the extract of Gratiola officinalis l. in rats with transplanted sarcoma 45 in experiment in vivo was conducted. Gratiola officinalis l. extract is received by patented method and is not toxic to animals. The study was conducted on 40 white male rats line Wistar weighing 150 ± 50 g. Animals were divided into 4 groups (10 rats per group): control group, comparison group with sarcoma without affecting, group with sarcoma with intramuscular and group with sarcoma with oral administration of the extract in a dosage of 110 mg/kg. The extract was administered intramuscularly or orally 72 hours after transplantation of sarcoma 45. The tumor volume and the weight of the animals were assessed daily. Results. The extract of leaves and flowers of Gratiola officinalis l. obtained by patented method has a strong antitumor activity, reducing the growth rate of the tumor and causing marked changes in the tumor, as well as providing stable anticachexia effect. Index of tumor weight inhibition was 70.6 % on average. Intramuscular administration was more effective in reducing of tumor growth, but less effectively increases the weight of animals than oral administration. In both administration methods Gratiola officinalis extract has no toxic effect on peripheral blood. We have previously found that the extract has antioxidant activity so that anticachexia effect is pathogenic, meaning it occurs by reducing toxicity. Conclusions. Gratiola officinalis extract has a broad spectrum of biological activity, in particular antitumor, anticachexia, it is not toxic, so it is advisable to investigate as a promising tool for the treatment of tumor diseases and cancer cachexia, and cachexia caused by other chronic diseases.

In the present work the effectiveness of antioxidants quercetine (a pure chemical) and Gratiola officinalis extract, which is obtained by a new method of extraction from plant material, is investigated on the model of photodynamic haemolysis that is a rather convenient method to monitor the rate of cell membranes oxidative destruction. The effect of these antioxidants on the rate of photodynamic haemolysis is considered as a measure of membranoprotective efficiency.