Evaluation of Phytochemistry and Antidiabetic Potential of an Astragalus Species: Astragalus kurdicus Boiss.

Astragalus kurdicus Boiss. roots are used in folk medicine for antidiabetic purposes. Different metabolites of Astragalus plants have a notable potential in antidiabetic activity via differing mechanisms. Herewith this study designed to assess the antidiabetic activity of Astragalus kurdicus, utilizing a range of diabetes-related in vitro methodologies and to investigate the chemical composition of the plant. According to the results of the activity tests, water extract (AKW) was the most active extract in PTP1B, DPP4, and α-amylase inhibition tests (87.17%, 82.4%, 91.49% respectively at 1 mg/ml). Total extract, AKM (85.63%), showed the highest AGEs inhibition activity. To test possible improvement effects of the extracts on diabetes through gut-microbiota, cell growth rates of three probiotic microorganisms were measured. AKM showed highest potential of prebiotic activity among tested extracts and caused higher biomass increase than standard prebiotics. Furthermore, flavonoid-rich extract was found to be mostly responsible for the high antioxidant activity. The highest saponin and astragalosides content were seen in AKB extract in HPTLC analysis. Among the measured saponins, the abundance of Astragaloside IV (27.41 µg/mg in AKM) was the highest in all fractions. Thus, for the first time, the antidiabetic activity of A. kurdicus was evaluated from various perspectives.

Uncovering chemical profiles, biological potentials, and protection effect against ECM destruction in H2 O2 -treated HDF cells of the extracts of Stachys tundjeliensis.

The genus Stachys L., one of the largest genera of the Lamiaceae family, is highly represented in Turkey. This study was conducted to determine the bio-pharmaceutical potential and phenolic contents of six different extracts from aerial parts of Stachys tundjeliensis. The obtained results showed that the ethanol extract exhibited the highest antioxidant activity in the antioxidant assays. Meanwhile, the ethanol extract displayed strong inhibitory activity against α-tyrosinase, the dichloromethane extract exhibited potent inhibition against butyrylcholinesterase, and the n-hexane extract against α-amylase. Based on ultra-high performance liquid chromatography coupled to high-resolution mass spectrometry analysis, more than 90 secondary metabolites, including hydroxybenzoic acid, hydroxycinnamic acid, and their glycosides, acylquinic acids, phenylethanoid glycosides, and various flavonoids were identified or tentatively annotated in the studied S. tundjeliensis extracts. It was observed that the application of S. tundjeliensis eliminated H2 O2 -induced oxidative stress. It was determined that protein levels of phospho-nuclear factor kappa B (NF-κB), receptor for advanced glycation endproducts, and activator protein-1, which are activated in the nucleus, decreased, and the synthesis of matrix metalloproteinase (MMP)-2 and MMP-9 also decreased to basal levels. Overall, these findings suggest that S. tundjeliensis contains diverse bioactive compounds for the development of nutraceuticals or functional foods with potent biological properties.

Unraveling the chemical profile, antioxidant, enzyme inhibitory, cytotoxic potential of different extracts from Astragalus caraganae.

Six extracts (water, ethanol, ethanol-water, ethyl acetate, dichloromethane, and n-hexane) of Astragalus caraganae were studied for their biological activities and bioactive contents. Based on high-performance liquid chromatography-mass spectrometry (HPLC-MS), the ethanol-water extract yielded the highest total bioactive content (4242.90 µg g-1 ), followed by the ethanol and water extracts (3721.24 and 3661.37 µg g-1 , respectively), while the least total bioactive content was yielded by the hexane extract, followed by the dichloromethane and ethyl acetate extracts (47.44, 274.68, and 688.89 µg g-1 , respectively). Rutin, p-coumaric, chlorogenic, isoquercitrin, and delphindin-3,5-diglucoside were among the major components. Unlike the dichloromethane extracts, all the other extracts showed radical scavenging ability in the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay (8.73-52.11 mg Trolox equivalent [TE]/g), while all extracts displayed scavenging property in the 2,2-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical scavenging assay (16.18-282.74 mg TE/g). The extracts showed antiacetylcholinesterase (1.27-2.73 mg galantamine equivalent [GALAE]/g), antibutyrylcholinesterase (0.20-5.57 mg GALAE/g) and antityrosinase (9.37-63.56 mg kojic acid equivalent [KAE]/g) effects. The molecular mechanism of the H2 O2 -induced oxidative stress pathway was aimed to be elucidated by applying ethanol, ethanol/water and water extracts at 200 µg/mL concentration to human dermal cells (HDFs). A. caraganae in HDF cells had neither a cytotoxic nor genotoxic effect but could have a cytostatic effect in increasing concentrations. The findings have allowed a better insight into the pharmacological potential of the plant, with respect to their chemical entities and bioactive contents, as well as extraction solvents and their polarity.

In Vitro Anti-Aging Potential Evaluation of Maclura pomifera (Rafin.) Schneider 80% Methanol Extract with Quantitative HPTLC Analysis.

Objectives: Maclura pomifera (Rafin.) Schneider is a widespread species all around the world, which is also frequently cultured for ornamental purposes. Previous studies revealed that M. pomifera fruits are rich in prenylated isoflavonoids, exhibit noteworthy biological activities, and have probable benefits, particularly, when applied topically. Considering that phenolic compounds are important sources in the development of anti-aging cosmetic products, this study investigated the anti-aging potential of M. pomifera 80% methanolic extract (MPM) by evaluating antioxidant and extracellular matrix degrading enzymes inhibiting activity. Materials and Methods: For this study, the inhibitory potential of 80% MPM against different enzymes associated with the aging process was evaluated. Given the unequivocal role of oxidative stress in aging, in vitro antioxidant tests were employed as well. Moreover, osajin was determined as the major bioactive isoflavonoid of the sample by high performance thin layer chromatography analysis. Results: Results of the mechanistically different antioxidant assays exhibited notable antioxidant potential of the extract. Inhibition potential of MPM against hyaluronidase, collagenase, and elastase enzymes, which are directly linked to acceleration of the aging process, was investigated and results revealed that MPM inhibited the aforementioned enzymes explicitly. MPM had notable phenolic and flavonoid content; 113.92 ± 2.26 mg gallic acid equivalent/g and 66.41 ± 0.74 mg QE/g, respectively. When total antioxidant capacity assays were considered, it is possible to suggest that MPM is a promising anti-aging agent. Conclusion: As a result, this study discloses that extracts of fruits of M. pomifera have significant anti-aging potential and may be used for this purpose.

An In-Depth Study on the Metabolite Profile and Biological Properties of Primula auriculata Extracts: A Fascinating Sparkle on the Way from Nature to Functional Applications.

The biological activity of the aerial part and rhizomes of Primula auriculata were assessed for the first time. The biological activities (antioxidant properties, enzyme inhibition, and AGE inhibition) as well as the phenolic and flavonoid contents of the ethyl acetate, ethanol, hydro-ethanol and water extracts of P. auriculata aerial parts and rhizomes were determined. Cell viability assays and gelatin zymography were also performed for MMP-2/-9 to determine the molecular mechanisms of action. The gene expression for MMPs was described with RT-PCR. The levels of various proteins, including phospho-Nf-κB, BCL-2, BAX, p-53, and cyclin D1 as well as RAGE were measured using Western blot analysis. The hydro-ethanol extract of the aerial part possessed the highest phenolic (56.81 mg GAE/g) and flavonoid (63.92 mg RE/g) contents. In-depth profiling of the specialized metabolites by ultra-high-performance liquid chromatography-high-resolution mass spectrometry (UHPLC-HRMS) allowed for the identification and annotation of 65 compounds, including phenolic acids and glycosides, flavones, flavonols, chalcones, dihydrochalcones, and saponins. The hydro-ethanol extract of the aerial parts (132.65, 180.87, 172.46, and 108.37 mg TE/g, for the DPPH, ABTS, CUPRAC, and FRAP assays, respectively) and the ethanol extract of the rhizomes (415.06, 638.30, 477.77, and 301.02 mg TE/g, for the DPPH, ABTS, CUPRAC, and FRAP assays, respectively) exhibited the highest free radical scavenging and reducing activities. The ethanol and hydro-ethanol extracts of both the P. auriculata aerial part and rhizomes exhibited higher inhibitory activity against acetylcholinesterase, while the hydro-ethanol extracts (1.16 mmol ACAE/g, for both the aerial part and rhizomes extracts) were more active in the inhibition of α-glucosidase. After the treatment of an HT-29 colorectal cancer cell line with the extracts, the apoptosis mechanism was initiated, the integrity of the ECM was remodeled, and cell proliferation was also taken under control. In this way, Primula extracts were shown to be potential drug sources in the treatment of colorectal cancer. They were also detected as natural MMP inhibitors. The findings presented in the present study appraise the bioactivity of P. auriculata, an understudied species. Additional assessment is required to evaluate the cytotoxicity of P. auriculata as well as its activity in ex vivo systems.

Nitrosylation of tissue transglutaminase enhances fibroblast migration and regulates MMP activation.

In wound healing, the TG2 enzyme plays a dual functional role. TG2 has been shown to regulate extracellular matrix (ECM) stabilization by its transamidase activity while increasing cell migration by acting as a cell adhesion molecule. In this process, nitric oxide (NO) plays a particularly important role by nitrosylation of free cysteine ​​residues on TG2, leading to the irreversible inactivation of the catalytic activity. In this study, transfected fibroblasts expressing TG2 under the control of the tetracycline-off promoter were treated with NO donor S-nitroso-N-acetyl penicillamine (SNAP) to analyze the interplay between NO and TG2 in the regulation of cell migration/invasion as well as TGF-ß1-dependent MMP activation. Our results demonstrated that inhibition of TG2 cross-linking activity by SNAP promoted the migration and invasion capacity of fibroblasts by hindering TG2-mediated TGF-ß1 activation. While the inhibition of TG2 activity by NO downregulated the biosynthesis and activity of MMP-2 and MMP-9, that of MMP-1a and MMP-13 was shown to be upregulated in a TGF-ß1-dependent manner under the same conditions. In the presence of SNAP, interaction of TG2 with its cell surface binding partners Integrin-ß1 and Syndecan-4 was reduced, which was paralleled by an increase in TG2 and PDGF association. These findings suggests that migratory phenotype of fibroblasts can be regulated by the interplay between nitric oxide and TG2 activity.

Influence of In Vitro Human Digestion Simulation on the Phenolics Contents and Biological Activities of the Aqueous Extracts from Turkish Cistus Species.

Oxidative stress is one of the significant precursors of various metabolic diseases such as diabetes, Parkinson's disease, cardiovascular diseases, cancer, etc. Various scientific reports have indicated that secondary plant metabolites play an important role in preventing oxidative stress and its harmful effects. In this respect, this study was planned to investigate the phenolic profile and antioxidant and antidiabetic potentials of the aqueous extracts from Turkish Cistus species by employing in vitro methods. In vitro digestion simulation procedure was applied to all extracts to estimate the bioavailability of their phenolic contents. Total phenolic, flavonoid, phenolic acid and proanthocyanidin contents were determined for all phases of digestion. In addition, changes in the quantity of the assigned marker flavonoids (tiliroside, hyperoside and quercitrin) were monitored by High-Performance Thin Layer Chromatography (HPTLC) analysis. The antioxidant activity potentials of the extracts were studied by various methods to reveal their detailed activity profiles. On the other hand, in vitro α-amylase and α-glucosidase enzymes and advanced-glycation end product (AGE) inhibitory activities of the extracts were determined to evaluate the antidiabetic potentials of extracts. The results showed that aqueous extracts obtained from the aerial parts of Turkish Cistus species have rich phenolic contents and potential antioxidant and antidiabetic activities; however, their bioactivity profiles and marker flavonoid concentrations might significantly be affected by human digestion. The results exhibited that total phenolic contents, antioxidant activities and diabetes-related enzyme inhibitions of the bioavailable samples were lower than non-digested samples in all extracts.

Comprehensive evaluation of two Astragalus species (A. campylosema and A. hirsutus) based on biological, toxicological properties and chemical profiling.

Astragalus L. (Fabaceae) is an important genus with numerous species having various traditional medicinal uses making them of interest for scientific investigations to ascertain their therapeutic benefits. In the present study, the quantitative polyphenolic profiles of methanolic extracts from different parts (leaves, flowers, and roots) of two endemic Astragalus species growing in Turkey, i.e. A. campylosema Boiss. and A. hirsutus Vahl were determined, along with their antioxidant and enzyme inhibitory properties. A. campylosema and A. hirsutus extracts showed varying total phenolic (25.80-40.60 and18.59-29.46 mg GAE/g, respectively) and total flavonoid (11.21-105.91 and 16.06-131.91 mg RE/g, respectively) contents. HPLC-MS/MS revealed rutin to be the predominant phenolic compound in all the extracts of A. campylosema and leaf extract of A. hirsutus (133.53-752.42 µg g-1), while hyperoside was the major one in the flower and root extracts of A. hirsutus (2014.07 and 123.13 µg g-1, respectively). In DPPH and ABTS assays, radical scavenging capacity was demonstrated by all extracts of A. campylosema (47.13-48.10 and 87.03-115.36 mg TE/g, respectively) and A. hirsutus (17.82-38.67 and 47.84-57.29 mg TE/g, respectively). Reducing activity was also displayed by the extracts in CUPRAC and FRAP assays (A. campylosema: 83.06-135.20 and 59.15-90.19 mg TE/g, respectively; A. hirsutus: 53.02-83.42 and 31.25-43.25 mg TE/g, respectively). All extracts were also found to act as metal chelators (12.32-21.45 mg EDTAE/g) and exhibited total antioxidant capacity ranging from 1.16 to 1.60 mmol TE/g, in phosphomolybdenum assay. Acetyl- and butyryl-cholinesterase inhibitory effects were observed by all the extracts of the two species (1.56-4.99 mg GALAE/g). Anti-hyperpigmentation potential by inhibiting tyrosinase (54.55-67.35 mg KAE/g) was reported as well. Carbohydrate hydrolyzing enzymes, amylase and glucosidase were also inhibited (0.22-1.03 mmol ACAE/g). Overall, A. campylosema extracts showed relatively better antioxidant and enzyme inhibitory potentials compared to A. hirsutus extracts. Strikingly, A. hirsutus extracts was found to have higher AGE inhibition activity than A. campylosema. Although the cytotoxic effect of three different organs obtained from A. campylosema and A. hirsutus increased depending on the dose (from 10 to 200 µg/mL), it was found that both plant extracts did not show a genotoxic effect at the highest concentration of 200 µg/mL. Indeed, data amassed from this current scientific work showed the two selected Astragalus species to be rich in bioactive polyphenols that could be responsible for the various pharmacological activities and hence demands to be further explored for their possible applications as natural health promoting agents.

Hypericum olympicum L. recovers DNA damage and prevents MMP-9 activation induced by UVB in human dermal fibroblasts.

ETHNOPHARMACOLOGICAL RELEVANCE: Hypericum olympicum L. (Hypericaceae) flowering aerial parts has been utilized in Turkish folk medicine as a remedy against inflamed skin problems. AIM OF THE STUDY: This study was designed to state the effect of H. olympicum on dermatological problems. For this purpose effect of the plant extract on the DNA strand break and matrix metalloproteinase (MMP)-9 activity of human dermal fibroblast (HDFs) cells irradiated with UVB as well as antioxidant activity potential were studied. MATERIALS AND METHODS: The methanolic extract of Hypericum olympicum (HOM) was prepared by maceration at room temperature. DNA damage and increased MMP-9 activity in HDFs were induced by UVB irradiation. The cell viability was measured by water-soluble tetrazolium salt (WST)-1 assay. The effects on DNA strand break was investigated by single gel electrophoresis (commonly known as Comet assay), while MMP-9 activity was observed by gelatin zymography assay. In vitro antioxidant tests were performed to indicate the effect on reactive oxygen species (ROS). The major metabolites were identified and their concentrations were measured by high performance thin layer chromatography (HPTLC). RESULTS: HOM was found to recover DNA damage dose-dependently. The enzymatic activity of MMP-9 was inhibited almost 100% by the treatment of 1.5 mg/mL of the extract. It also enhanced cell proliferation in those cells, and also it was shown to possess antioxidant activity. The major metabolites of HOM were identified as chlorogenic acid and quercetin glycosides (rutin, hyperoside, isoquercitrin). CONCLUSION: Experimental studies have proven the traditional use of Hypericum olympicum in inflamed skin problems acting by inhibition of the inflammatory pathway and recovery of DNA damage induced experimentally.