ИСТИНА

Duranta repens is a common plant in Southern China and is often used in traditional medicine to treat bruises (fig. 1). Fruits and leaves of Duranta were used for the treatment of malaria and abscess (Takeda et al. 1995). Recent data demonstrate that Duranta repens extracts exhibit variable effects on human and animal cells, including cytotoxicity, antioxidant activity, antidiabetic effect, activation of insulin secretion, etc (Khanal and Patil, 2021; da Silva et al, 2016). We aimed to investigate the effect of ethanol extracts obtained from leaves (I) and callus cultures (II) of Duranta repens on human immortalized keratinocytes (HaCaT) and primary human adipose-derived stem cells (hASCs). Both cell lines can be obtained from human skin (fig. 2). Using MTT assay, we evaluated the metabolic activity (cell viability) of HaCaT and hASCs cells, growing for 24 and 48 h in the presence of leaf and callus extracts (fig. 3) at concentrations varying from 12,5 µg/ml to 125 µg/ml (fig. 4). It was found that primary hASCs were more sensitive to the incubation with both types of extracts than immortalized HaCaT cells. A decrease in the metabolic activity of hASCs was observed after incubation for 24 and 48 hours with 125 µg/ml of callus extract and 25 µg/ml – 125 µg/ml of leaf extract (fig. 4). The changes in metabolic activity of HaCaT cells were detected only after 24 h of treatment: 125 µg/ml of callus extract increased the metabolic activity and 125 µg/ml of leaf extract decreased it (fig. 4). To study the long-term effects, we chose a concentration of both extracts that does not affect cell viability (25 µg/ml). Using flow cytometry, we demonstrated that both extracts does not affect the course of cell cycle in HaCaT cells (fig. 5). It has been suggested that Duranta repens extracts affect the lipid metabolism in rats (Khanal and Patil, 2021), therefore we investigated the effect of extracts on lipid droplet formation during adipogenic differentiation of hASCs (14 and 21 days) (fig. 7). Before that, we showed that the hASC phenotype corresponds to the phenotype of MSC (fig. 6). We obtained preliminary data on the number of adipocytes after the incubation with callus extract (fig. 7). The leaf extract stimulated the appearance of multiple large intracellular vesicles (not lipid) (4, 14, 21 d) (fig. 7). Conclusions: Duranta repens callus and leaf extracts demonstrate heterogeneity of effects on cultured human skin cells.