Introduction. Geroprotectors are biologically active substances that inhibit the aging process. Many plant species are natural geroprotectors. For instance, Thymus vulgaris and Trifolium pratense are callus cultures with strong geroprotective properties. Study objects and methods. The present research featured T. vulgaris and T. pratense grown in vitro on various nutrient media. Their extracts were obtained by aqueous-alcoholic extraction using the following parameters: water-ethanol solvent Se = 30, 50, and 70 %; temperature Te = 30, 50, and 70°C; time τe = 2, 4, and 6 h. The quantitative and qualitative analysis was based on high-performance liquid mass spectrometry, gas mass spectrometry, and thin-layer chromatography. Results and discussion. The optimal extraction parameters for T. vulgaris were τe = 4 h, Te = 50°C, Se = 70 %, for T. pratense – τe = 6 h, Te = 70°C, Se = 70 %. The chromatography detected flavonoids, phenylpropanoids, simple phenols, higher fatty acids, mono- and sesquiterpenes, and aliphatic hydrocarbons. T. vulgaris appeared to have the highest content of thymol (23.580 ± 1.170 mg/mL); its thymol, apigenin, gallic, chlorogenic, and caffeic components demonstrated geroprotective properties. The extract of T. pratense possessed the highest rutin content (10.05 ± 0.35 mg/mL), and it owed its geroprotective characteristics to rutin, chlorogenic and p-coumaric acids. Conclusion. The callus cultures of T. vulgaris and T. pratense proved to be promising sources of geroprotective biologically active substances.
Trans-cinnamic acid is a phenolic compound with a wide range of bioactive properties, including antioxidant and antibacterial effects. It also has high potential in the food and cosmetic industries. We aimed to isolate trans-cinnamic acid from the Baikal skullcap (Scutellaria baicalensis) and study its geroprotective activity on Caenorhabditis elegans nematodes used as a model organism. Our study objects included the S. baicalensis root culture and its extract, trans-cinnamic acid isolated from the extract, and C. elegans nematodes. Trans-cinnamic acid was isolated by high-performance liquid chromatography. The acid’s geroprotective activity was studied by evaluating its effect at concentrations of 10, 50, 100, and 200 μmol/L on the lifespan, stress resistance, and reproductivity of C. elegans. For the lifespan study, the nematodes were cultivated at 20°C for 61 days. To assess their resistance to oxidative stress, 15 μL of 1M paraquat was added to each well of the plate. Thermal stress resistance was determined by raising the temperature to 33°C. For the reproductivity study, the nematodes were cultivated in the S-medium with the addition of Escherichia coli OP50 and trans-cinnamic acid at required concentrations for 72 h. The maximum increase in lifespan (9.8%) was observed in the nematodes treated with 50 μmol/L of trans-cinnamic acid. Under oxidative stress, all the concentrations of trans-cinnamic acid increased the survival of nematodes, while under thermal stress, trans-cinnamic acid reduced the percentage of surviving nematodes. At a concentration of 100 μmol/L, trans-cinnamic acid increased the nematodes’ reproduction by 1.48 times. Based on our data, trans-cinnamic acid isolated from S. baicalensis can be recommended as a bioactive compound with geroprotective activity. However, further research is needed on other model organisms with detailed toxicity studies.
Medicinal plants are sources of natural antioxidants. Acting as reducing agents, these substances protect the human body against oxidative stress and slow down the aging process. We aimed to study the effects of bioactive substances isolated from medicinal plants on the lifespan of Caenorhabditis elegans L. used as a model organism. High-performance liquid chromatography was applied to isolate bioactive substances from the extracts of callus, suspension, and root cultures of meadowsweet (Filipendula ulmaria L.), ginkgo (Ginkgo biloba L.), Baikal skullcap (Scutellaria baicalensis L.), red clover (Trifolium pretense L.), alfalfa (Medicágo sativa L.), and thyme (Thymus vulgaris L.). Their effect on the lifespan of C. elegans nematodes was determined by counting live nematodes treated with their concentrations of 10, 50, 100, and 200 µmol/L after 61 days of the experiment. The results were recorded using IR spectrometry. The isolated bioactive substances were at least 95% pure. We found that the studied concentrations of trans-cinnamic acid, baicalin, rutin, ursolic acid, and magniferin did not significantly increase the lifespan of the nematodes. Naringenin increased their lifespan by an average of 27.3% during days 8–26. Chlorogenic acid at a concentration of 100 µmol/L increased the lifespan of C. elegans by 27.7%. Ginkgo-based kaempferol and quercetin, as well as red clover-based biochanin A at the concentrations of 200, 10, and 100 µmol/L, respectively, increased the lifespan of the nematodes by 30.6, 41.9, and 45.2%, respectively. The bioactive substances produced from callus, root, and suspension cultures of the above medicinal plants had a positive effect on the lifespan of C. elegans nematodes. This confirms their geroprotective properties and allows them to be used as anti-aging agents.
Introduction. Wild-crafting leads to the local extinction of many medicinal plants that are rich in phenolic substances. In vitro cultivation of cells and organs of higher plants can be the optimal solution to this problem. The research objective was to study the biosynthetic activity of in vitro extracts of wild Siberian plants. Study objects and methods. The study featured callus, cell suspension, and hairy root extracts of such Siberian medicinal plants as Eleutherococcus senticosus, Codonopsis pilosula, Platanthera bifolia, and Saposhnikovia divaricata. They were obtained by in vitro cultivation using modified nutrient media of Murashige and Skoog and Gamborg. The content of secondary metabolites was studied using the methods of thin-layer and high-performance liquid chromatography. A set of in vitro experiments tested the antioxidant and antimicrobial activity of the extracts. Results and discussion. All the samples demonstrated a high content of secondary metabolites of phenolic nature. Flavonoglycosides, apigenin, and rutin were found to be the predominant biologically active substances in the callus extracts. Flavonoglycosides dominated in the suspension extracts. The root extracts contained more caffeic acid, rutin, ecdysteroids, quercetin, apigenin, cardiofolin, and coleofolide than the callus and suspension cultures. The list of prevailing secondary metabolites in the root extracts included rutin, apigenin, coleofolide, and quercetin. All the extracts showed antimicrobial and antioxidant activity. Conclusion. All the extracts demonstrated good antioxidant and antimicrobial properties. Therefore, they can be used for the production of pharmaceuticals and biologically active food supplements as they can be helpful against infectious diseases, as well as oncological, cardiovascular, and neurodegenerative diseases linked to oxidative stress.
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