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. 2022 Aug 30:2022:8368717.
doi: 10.1155/2022/8368717. eCollection 2022.

Natural Deep Eutectic Extracts of Propolis, Sideritis scardica, and Plantago major Reveal Potential Antiageing Activity during Yeast Chronological Lifespan

Bela Vasileva  1 , Dessislava Staneva  1 , Tsvetinka Grozdanova  2 , Hristo Petkov  2 , Boryana Trusheva  2 , Kalina Alipieva  2 , Milena Popova  2 , George Miloshev  1 , Vassya Bankova  2 , Milena Georgieva  1
Affiliations

Natural Deep Eutectic Extracts of Propolis, Sideritis scardica, and Plantago major Reveal Potential Antiageing Activity during Yeast Chronological Lifespan

Bela Vasileva et al. Oxid Med Cell Longev. .

Abstract

Nowadays, the environmentally friendly approach to everyday life routines including body supplementation with pharma-, nutraceuticals and dietary supplements gains popularity. This trend is implemented in pharmaceutical as well as cosmetic and antiageing industries by adopting a newly developed green chemistry approach. Following this trend, a new type of solvents has been created, called Natural Deep Eutectic Solvents (NADES), which are produced by plant primary metabolites. These solvents are becoming a much better alternative to the already established organic solvents like ethanol and ionic liquids by being nontoxic, biodegradable, and easy to make. An interesting fact about NADES is that they enhance the biological activities of the extracted biological compounds. Here, we present our results that investigate the potential antiageing effect of CiAPD14 as a NADES solvent and three plant extracts with it. The tested NADES extracts are from propolis and two well-known medicinal plants-Sideritis scardica and Plantago major. Together with the solvent, their antiageing properties have been tested during the chronological lifespan of four Saccharomyces cerevisiae yeast strains-a wild type and three chromatin mutants. The chromatin mutants have been previously proven to exhibit characteristics of premature ageing. Our results demonstrate the potential antiageing activity of these NADES extracts, which was exhibited through their ability to confer the premature ageing phenotypes in the mutant cells by ameliorating their cellular growth and cell cycle, as well as by influencing the activity of some stress-responsive genes. Moreover, we have classified their antiageing activity concerning the strength of the observed bioactivities.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Cell culture growth of WT, hho1delta, arp4, and arp4 hho1delta without and with the presence of CiAPD14 NADES solvent and the three extracts. Cells were cultivated for the period of 9 days in SC media, supplemented with the studied NADES solvent and extracts. The control cells were not supplemented. At four time points: 4 h, 24 h, 72 h, and 9 d, aliquots were taken from the cell cultures and were analyzed spectrophotometrically. The optical density was measured in arb. units. Data are MEAN ± SD. Statistically significant differences between the growth of supplemented yeast cells compared to nonsupplemented are given with asterisk (∗); (a) WT, where ∗ means p < 0.05 for CiAPD14 P. major; (b) hho1delta, where ∗ means p < 0.05 for CiAPD14 propolis; (c) arp4, where ∗ means p < 0.05 for CiAPD14 propolis; (d) arp4 hho1delta, where ∗ means p < 0.05 for CiAPD14.
Figure 2
Figure 2
CLS of WT, hho1delta, arp4, and arp4 hho1delta in the presence of CiAPD14 and the three extracts. Cells were cultivated for the period of 9 days in SC media supplemented with the studied NADES solvent and extracts. At three time points: 24 h, 72 h, and 9 d 100, cells were taken from the cell cultures and were seeded on rich solid YPD media. Cells were allowed to recover under optimal conditions (30°C). The cell survival at the 24th-time point was assumed as 100%, and the survival of cells at all other time points were calculated as a percentage of it. (a) CLS of the four studied strains without any treatment; (b) CLS of the WT cells, control and supplemented with CiAPD14 and extracts; (c) CLS of hho1delta mutant cells, control and supplemented with CiAPD14 and extracts; (d) CLS of arp4 mutants; (e) CLS of the double arp4 hho1delta control cells and in the presence of CiAPD14 and extracts. Data are MEAN ± SD. Statistically significant differences are marked with ∗ where p < 0.05.
Figure 3
Figure 3
Cell cycle analysis of the four studied strains WT, hho1delta, arp4, and arp4 hho1delta in the presence of CiAPD14 and the three extracts during their CLS. Cells were cultivated during the CLS for nine days. At four time points: 4 h, 24 h, 72 h, and 9 d, aliquot cells were taken from the cell cultures and were analyzed by FACS after propidium iodide staining. (a) WT strain FACS data quantitation (bar chart) and representative histograms (below) for each time point; (b) hho1delta strain FACS data quantitation (up) and representative histograms for each time point; (c) arp4 FACS data quantitation (up, bar chart) and representative histograms for each time point; (d) arp4 hho1delta FACS data quantitation (up) and representative histograms for each time point. Data are represented as % of cells in each phase of the cell cycle.
Figure 4
Figure 4
Gene expression analysis of the CDC28 gene of treated with CiAPD14, CiAPD14 propolis, CiAPD14 S. scardica, and CiAPD14 P. major extract yeast cells. Total RNA was isolated from aliquots of the yeast cultures and converted to cDNA. cDNA was used as a template in RT-qPCR experiments to analyze the expression of CDC28. ACT1 was expression the reference gene. (a) WT, (b) hho1delta, (c) arp4, and (d) arp4 hho1delta. The results were calculated by the ΔΔCT method using specialized Rotor-Gene 6000 software. Data are MEAN ± SD. Statistically significant differences are marked with ∗ where p < 0.05.
Figure 5
Figure 5
Gene expression analysis of the RAD9 gene of supplemented with CiAPD14, CiAPD14 propolis, CiAPD14 S. scardica, and CiAPD14 P. major extract yeast cells during their CLS. Total RNA was isolated from aliquots of the yeast cultures and converted to cDNA. cDNA was used as a template in RT-qPCR experiments to analyze the expression of RAD9. ACT1 was the reference gene. (a) WT control cells, (b) hho1delta mutant yeast cells, (c) arp4 mutants, and (d) arp4 hho1delta double mutant cells. The results were calculated by the ΔΔCT method using the specialized Rotor-Gene 6000 software. Data are MEAN ± SD. Statistically significant differences are marked with ∗ where p < 0.05.
Figure 6
Figure 6
Combined general antiageing effects of CiAPD14 and its extracts for all the studied S. cerevisiae strains during their CLS.
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