No abstract

Drosophila melanogaster mutants with deficient kynurenine (KYN) formation from tryptophan (TRP) have longer life span than wild type flies. Administration of alpha-methyl-TRP and 5-methyl-TRY, the inhibitors of TRP-KYN metabolism, prolonged life span in wild-type flies. Both inhibitors are not available for human use. Berberine, an isoquinoline alkaloid isolated from Berberis aristata, is known as the herb widely used in traditional Chinese and Indian medicine. Berberin is a strong inhibitor of the enzyme catalyzing TRP conversion into KYN. Considering this particular feature we investigated the effect of berberine on life- and health-span in wild-type Drosophila melanogaster. The results of our study showed that Berberine extended mean, median and maximum life span of female flies. Berberine did not affect the number of pupae of filial generation and decreased their lethality. Berberine increased locomotor activity (vertical climbing). The results of the study suggest that berberine prolongs life- and improves health-span of Drosophila melanogaster. Berberine might be a candidate drug for prevention and treatment of aging and aging-associated medical and psychiatric disorders.

Exposure to high sugar diet (HSD) serves as an experimental model of insulin resistance (IR) and type 2 diabetes (T2D) in mammals and insects. Peripheral IR induced by HSD delays emergence of pupae from larvae and decreases body weight of Drosophila imago. Understanding of mechanisms of IR/T2D is essential for refining T2D prevention and treatment strategies. Dysregulation of tryptophan (TRP) – kynurenine (KYN) pathway was suggested as one of the mechanisms of IR development. Rate-limiting enzyme of TRP – KYN pathway in Drosophila is TRP 2,3-dioxygenase (TDO), an evolutionary conserved ortholog of human TDO. In insects TDO is encoded by vermilion gene. TDO is not active in vermilion mutants. In order to evaluate the possible impact of deficient formation of KYN from TRP on the inducement of IR by HSD, we compared the effect of HSD in wild type (Oregon) and vermilion mutants of Drosophila melanogaster by assessing the time of white pupae emergence from larva and body weight of imago. Delay of emergence of pupae from larvae induced by high sucrose diet was less pronounced in vermilion (1.4 days) than in Oregon flies (3.3 days) in comparison with flies maintained on standard diet. Exposure to high sucrose diet decreased body weight of Oregon (but not vermilion) imago. Attenuation of high sucrose diet–induced IR/T2D in vermilion flies might depend on deficiency of TRP – KYN pathway. Besides IR/T2D, HSD induces obesity in Drosophila. Future studies of HSD-induced obesity and IR/T2D in TDO deficient vermilion mutants of Drosophila might help to understand the mechanisms of high association between IR/T2D and obesity. Modulation of TRP – KYN metabolism might be utilized for prevention and treatment of IR/T2D.

We have observed that berberine prolonged life span and improved viability of pupae and climbing activity of imagoes of wild-type Drosophila melanogaster maintained at 23°C. As a continuation of our studies of berberine effect on life span, we were interested to evaluate the effect of berberine of life span in flies maintained at a higher temperature (28°C) known to accelerate aging in wild type flies. Considering that genetically or pharmacologically induced deficiency of TRP conversion into KYN prolonged life span in a Drosophila model, we compared the effects of berberine, a powerful inhibitor of kynurenine (KYN) formation from tryptophan (TRP), on life span in wild type and in Drosophila melanogaster mutants (vermilion) with deficient TRP-KYN metabolism maintained at 23°C and 28°C. High (28°C) ambient temperature decreased life span in both wild type and vermilion flies. Aging accelerating effect of high temperature was more pronounced in Oregon than in vermilion flies (−60% vs. −40% decrease of mean life span, resp). Berberine attenuated the aging-accelerating effect of high temperature. Effect of berberine was more pronounced in Oregon (+46%) than in vermilion (+22%) flies. The obtained data suggested the possible involvement of TRP-KYN metabolism in the aging-acceleration effect of the high temperature and in protective effect of berberine.

Intracellular microelectrophoresis was applied to investigate the electrokinetic properties of human buccal epithelium cell nuclei after exposure of cells to microwaves of wavelengths of 8 mm (f = 37.5 GHz) and 16 mm (f = 18.75 GHz) at a surface power density of 0.2 mW/cm(2). Irradiated or nonirradiated cells were suspended in a flat microelectrophoretic chamber and exposed to an electric field of 15 V/cm at a current flow of 0.1 mA. The cells, whose nuclei altered their intracellular location towards the anode of the externally applied electric field, were considered to have negatively charged nuclei. The percentage of cells with electrophoretically movable nuclei was determined as the value of electronegativity of cell nuclei (ENN). Microwaves induced changes of ENN during irradiation of 15-60 s. If cells of a donor had an elevated initial level of ENN, it decreased during irradiation. On the contrary, if cells of another donor had a low initial ENN level, irradiation induced ENN increase. No significant difference between the action of microwaves of wavelengths of 8 mm and 16 mm was found. However, microwave irradiation caused an increase in membrane permeability for the in vivo dye indigo Carmine in cells of all donors irrespectively of the initial levels they showed. This suggests that electrokinetic properties of nuclei in cells do not only depend on cell membrane permeability.