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.

Phytochemical analysis, antioxidant and hypoglycemic activities of a methanol extract from Stachys brachyclada de Noé ex Coss. leaves.

Stachys brachyclada de Noé ex Coss. (Lamiaceae) is a quite rare medicinal plant endemic to the Mediterranean basin. In this study, seven secondary metabolites from a methanol extract of its leaves have been isolated and identified by a combination of chromatographic and spectroscopic methods (1D and 2D NMR experiments and ESIMS analysis). They include one ethyl 4-hydroxybenzoate (1), three acylated flavone glycosides (2-4), one diapigenin derivative (5) and two flavone aglycones (6-7). Stachysetin (5) was found the major compound of the extract (74.0 mg/g of dry matter). Moreover, the produced extract showed the ability in inhibiting the α-glucosidase enzyme (IC50 = 13.7 µg/mL), in quenching the radical 1,1-diphenyl-2-picrylhydrazyl (EC50 = 74.6 µg/mL), and in reducing the intracellular oxidative stress level in Human Dermal Fibroblast (64% inhibition at 50 µg/mL).

Isolation and Purification of Harpagogenin as an Nrf2-ARE Activator from the Tubers of Chinese Artichoke (Stachys sieboldii).

Chinese artichoke tuber (Stachys sieboldii Miq.) is used as an herbal medicine as well as edible food. This study examined the effect of the Chinese artichoke extracts on the nuclear factor erythroid 2-related factor 2 (Nrf2)-antioxidant response element (ARE) pathway that induces the expression of antioxidant enzymes to explore its novel characteristics. Hot water extracts exhibited relatively high ARE activity. ARE activity was observed in two fractions when the hot water extracts were separated in the presence of trifluoroacetic acid using HPLC. Conversely, the highly active fraction disappeared when the hot water extracts were separated in the absence of trifluoroacetic acid. These results indicate that acidic degradation produces active ingredients. The structural analysis of the two active fractions identified harpagide, which is an iridoid glucoside, and harpagogenin. In vitro experiments revealed that harpagide was converted into harpagogenin under acidic conditions and that harpagogenin, but not harpagide, had potent ARE activity. Therefore, this study identified harpagogenin, which is an acid hydrolysate of harpagide, as an ARE activator and suggests that Nrf2-ARE pathway activation by Chinese artichoke contributes to the antioxidative effect.

The chemical composition of the aerial parts of Stachys spreitzenhoferi (Lamiaceae) growing in Kythira Island (Greece), and their antioxidant, antimicrobial, and antiproliferative properties.

The Stachys L. genus has been used in traditional medicine to treat skin inflammations, stomach disorders, and stress. The aim of this study was to investigate the chemical profile and biological activity of the methanolic extract of Stachys spreitzenhoferi Heldr. (Lamiaceae) aerial parts, collected on the island of Kythira, South Greece. The analysis by liquid chromatography coupled with electrospray ionization and high-resolution mass spectrometry [LC-(-)ESI/HRMSn] of the methanol extract revealed the occurrence of thirty-six compounds - flavonoids, phenylethanoid glycosides, iridoids, quinic acid derivatives, aliphatic alcohol glycosides, and oligosaccharides - highlighting the substantial presence, as main peaks, of the iridoid melittoside (2) along with flavonoid compounds such as 4'-O-methylisoscutellarein mono-acetyl-diglycoside/chrysoeriol mono-acetyl-diglycoside (24), trimethoxy- (35) and tetramethoxyflavones (36). This extract was tested for its antimicrobial properties against Gram-positive and negative pathogenic strains. The extract was not active against Gram-negative bacteria tested, but it possessed a good dose-dependent antimicrobial activity towards S. aureus (MIC: 1.0 mg/mL) and L. monocytogenes (MIC: 1.0 mg/mL) Gram-(+) strains. Furthermore, this extract has been tested for its possible antioxidant activity in vitro. In particular, it has been shown that these molecules cause a decrease in DPPH, ABTS, and H2O2 radicals. The extract of S. spreitzenhoferi exhibited anti-DPPH activity (IC50: 0.17 mg/mL), anti-H2O2 activity (IC50: 0.125 mg/mL), and promising antiradical effect with an IC50 value of 0.18 mg/mL for anti-ABTS activity. S. spreitzenhoferi extract caused a decrease in ROS (at the concentration of 200 µg/mL) and an increase in the activity of the antioxidant enzymes SOD, CAT, and GPX in OZ-stimulated PMNs. Furthermore, it exhibited antiproliferative activity against acute myeloid leukemia (U937 cell), causing 50% of cell death at the 0.75 mg/mL.

Liposome Extract of Stachys pilifera Benth Effectively Improved Liver Damage due to Bile Duct Ligation Rats.

Herbal medicines harbor essential therapeutic agents for the treatment of cholestasis. In this study, we have assessed the anticholestatic potential of Stachys pilifera Benth's (SPB's) hydroalcoholic extract encapsulated into liposomes using bile duct ligation- (BDL-) induced hepatic cholestasis in rats. Aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), malondialdehyde (MDA), total thiol (T-SH) content, protein carbonyl (PCO), total bilirubin (TBIL), albumin (ALB), and nitric oxide (NO) metabolite levels were measured in either liver tissue or plasma to assess liver damage. Moreover, expression of proinflammatory cytokines (IL-1ß and TNF-α) and liver fibrosis markers (TGF-ß and SM-α) which are driving forces of many liver disorders was also determined. The activity of AST, ALT, and ALP was significantly enhanced in the BDL group in comparison to the control group; however, treatment with liposomal (SPB) hydroalcoholic extract significantly reduced AST and ALT's activity. Increases in MDA, TBIL, and NO levels and T-SH content due to BDL were restored to control levels by liposomal (SPB) hydroalcoholic extract treatment. Similarly, hepatic and plasma oxidative marker MDA levels, significantly enhanced by BDL, were significantly decreased by liposomal (SPB) hydroalcoholic extract treatment. Moreover, histopathological findings further demonstrated a significant decrease in hepatic damage in the liposomal (SPB) hydroalcoholic extract-treated BDL group. In addition, liposomal (SPB) hydroalcoholic extract treatment decreased the liver expression of inflammatory cytokines (IL-1ß, TNF-α) and liver fibrosis markers (TGF-ß and SM-α). Since liposomal (SPB) hydroalcoholic extract treatment alleviated the BDL-induced injury of the liver and improved the hepatic structure and function more efficiently in comparison to free SPB hydroalcoholic extract, probable liposomal (SPB) hydroalcoholic extract exhibits required potential therapeutic value in protecting the liver against BDL-caused oxidative injury.

PVA based nanofiber containing cellulose modified with graphitic carbon nitride/nettles/trachyspermum accelerates wound healing.

Today, bacterial cellulose has received a great deal of attention for its medical applications due to its unique structural properties such as high porosity, good fluid uptake, good strength, and biocompatibility. This study aimed to fabricate and study bacterial cellulose/graphitic carbon nitride/nettles/trachyspermum nanocomposite by immersion and PVA/BC/g-C3 N4 /nettles/trachyspermum nanofiber by electrospinning method as a wound dressing. The g-C3 N4 and g-C3 N4 solution were synthesized and then were characterized using Fourier transform infrared, X-ray diffraction, Zeta Potential, and scanning electronic microscope analyzes. Also, the antibacterial properties of the synthesized materials were proved by gram-positive and gram-negative bacteria using the minimum inhibitory concentration method. Besides, the toxicity, migration, and cell proliferation results of the synthesized materials on NIH 3T3 fibroblasts were evaluated using MTT and scratch assays and showed that the BC/PVA/g-C3 N4 /nettles/trachyspermum composite not only had no toxic effect on cells but also contributed to cell survival, cell migration, and proliferation has done. To evaluate the mechanical properties, a tensile strength test was performed on PVA/BC/g-C3 N4 /nettles/trachyspermum nanofibers, and the results showed good strength of the nanocomposite. In addition, in vivo assay, the produced nanofibers were used to evaluate wound healing, and the results showed that these nanofibers were able to accelerate the wound healing process so that after 14 days, the wound healing percentage showed 95%. Therefore, this study shows that PVA/BC/g-C3 N4 /nettles/trachyspermum nanofibers effectively inhibit bacterial growth and accelerate wound healing.

Chemical composition, antioxidant, and anti-diabetic activities of ethyl acetate fraction of Stachys riederi var. japonica (Miq.) in streptozotocin-induced type 2 diabetic mice.

This work analysed the chemical composition, antioxidant, and enzyme inhibitory activities of solvent extract (SJ-ME) and fractions (SJ-HF, SJ-EAF, and SJ-MF) of the Stachys riederi var. japonica (Miq.) (SJ). Furthermore, the effect of SJ-EAF in STZ induced type 2 diabetic mice was examined. Among the samples, SJ-EAF exhibited a lower IC50 concentration of 64.2 ± 0.48 µg/mL for DPPH and 82.6 ± 0.09 µg/mL for ABTS+. The SJ-EAF concentration of 2.89 ± 0.03 µg and 2.27 ± 0.98 µg was equivalent to 1 µg of acarbose mediated enzyme inhibitory effect against α-amylase and α -glucosidase, respectively. The SJ-EAF did not show cytotoxicity (<80%) to NIH3T3 nor HepG2 cells but enhanced the glucose uptake in the IR-HepG2. LC-MS/MS of SJ-EAF showed the presence of a total of 16 compounds. Among the identified compounds, rosmarinic acid, caffeic acid, oleanolic acid, and ursolic acid showed high catalytic activity of α-amylase and α-glucosidase. The treatments of SJ-EAF restored the level of blood glucose, body weight, insulin, HDL and mRNA level of IRS1, GLUT2, GLUT4 and Akt whereas it reduced the excess elevation of total cholesterol, total triglycerides, LDL, AST, ALT, ALP, BUN, and creatinine in STZ induced diabetic mice. Overall, the present study concluded that the SJ-EAF exhibited promising antidiabetic activity.

Chemical composition, antimicrobial, antioxidant and cytotoxic activity of the essential oil from the leaves of Stachys pilifera Benth.

Stachys pilifera Benth is an endemic species of Stachys family found in Iran with a wide application as an herbal tea. The objective of this research was to evaluate the antimicrobial, antioxidant and cytotoxic activity of the essential oil from the aerial parts of S. pilifera. Essential oil (EO) composition analysis showed that cis-Chrysanthenyl acetate (24.9%), viridiflorol (18.3%), trans-Caryophyllene (9.8%), caryophyllene oxide (4.6%), α-terpineol (3.3%) and linalool (3.1%) were the most abundant components. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the EO showed a higher antimicrobial activity against Gram-positive bacteria (Bacillus cereus and Staphylococcus aureus) than Gram-negative organisms (Escherichia coli, Shigella sonnei, Pseudomonas aeruginosa and Salmonella enterica subsp. Enterica). The antioxidant activity of EO was studied using DPPH, FRAP and ß-carotene/linoleic acid assays. IC50 for the DPPH, FRAP and ß-carotene/linoleic acid tests were 23.2, 28.7 and 16.1 µg/mL, respectively, that it was higher than the results for BHT (P ≤ 0.05). The cytotoxic activity of the EO was evaluated using HT29 and HUVEC cells and it was observed that by increasing in EO concentration from 0.026 to 19.4 ug/mL, the viability of the cells for HT29 and HUVEC reduced to 6.8 and 7.1%, respectively. The results from this study suggest the possibility to use the essential oils from S. pilifera Benth as a natural preservative in processed or packaged food due to its high antibacterial, antioxidant and cytotoxic activities.

The Effect of Stachy sieboldii MIQ. Supplementation on Modulating Gut Microflora and Cytokine Expression in Mice.

AIMS AND OBJECTIVES: The intake of Stachys sieboldii MIQ. has been associated with relieving inflammation and maintaining optimal gut health function. We investigated the diversity and composition of microflora in feces of S. sieboldii MIQ.-fed mice. In addition, we evaluated the production of major cytokines (Interleukin-6 and -10) related to inflammation and fatty acid composition of several tissues. MATERIALS AND METHODS: 16S ribosomal DNA sequencing-based microbiome taxonomic profiling analysis was performed using EzBioCloud data base. The total RNA from the mesenteric lymph node was isolated and then synthesized with prime script 1st strand cDNA synthesis kit. Quantitative real-time PCR was performed on cDNA samples using the SYBR™ Green PCR Master Mix. RESULTS: Mice fed on S. sieboldii MIQ. showed significantly reduced counts of aerobic and coliform in the feces compared with control. 16S rDNA sequencing analysis of fecal samples showed that supplementation with S. sieboldii MIQ. increased beneficial intestinal microflora (Ruminococcaceae and Akkermansia muciniphila) and decreased the community of harmful microflora (Enterobacteriaceae, including Escherichia coli and Bacteroides sp.) in feces compared with that in the control (P<0.05 for all). Mice showed a significantly lower mRNA expression of cytokines IL-6 and IL-10 in mesenteric lymph node compared with that in control (P<0.05). The fecal fatty acid composition in the S. sieboldii MIQ. group showed a higher percentage of 6:0 and 18:2n-6 compared with that in the control group (P<0.05). The percentages of 6:0 and 20:3n-6 fatty acids were also significantly higher in the intestines of S. sieboldii MIQ. group (P<0.05). No differences were revealed between the two groups in terms of the percentages of total saturated, monounsaturated, n-6 and n-3 polyunsaturated fatty acids found in feces and tissues. CONCLUSION: The present results showed that supplementation of mice with S. sieboldii MIQ. increased beneficial gut microflora and decreased harmful microflora. Moreover, lower mRNA expression of pro-inflammatory cytokine IL-6, and anti-inflammatory cytokine IL-10 in the mesenteric lymph node of supplemented mice might be associated with the lower abundances of harmful fecal microflora.

Valorisation of stachysetin from cultivated Stachys iva Griseb. as anti-diabetic agent: a multi-spectroscopic and molecular docking approach.

Stachys species are considered as important medicinal plants with numerous health benefit effects. In continuation of our research on the Greek Stachys species, the chemical profile of the aerial parts of cultivated S. iva Griseb. has been explored. The NMR profiles of the plant extract/infusion were used to guide the isolation process, leading to the targeted isolation of seventeen known compounds. The rare acylated flavonoid, stachysetin, was isolated for the third time from plant species in the international literature. Identification of the characteristic signals of stachysetin in the 1D 1H-NMR spectrum of the crude extract was presented. In order to evaluate the potential of the identified chemical space in Stachys to bear possible bioactivity against diabetes, we performed an in silico screening against 17 proteins implicated in diabetes, as also ligand based similarity metrics against established anti-diabetic drugs. The results capitalized the anti-diabetic potency of stachysetin. Its binding profile to the major drug carrier plasma protein serum albumin was also explored along with its photophysical properties suggesting that stachysetin could be recognized and delivered in plasma through serum albumin and also could be tracked through near-infrared imaging. Communicated by Ramaswamy H. Sarma.

Isolation of bioactive components with soluble epoxide hydrolase inhibitory activity from Stachys sieboldii MiQ. by ultrasonic-assisted extraction optimized using response surface methodology.

Stachys sieboldii MiQ (SSM) is an important food and medicinal herb in Korea, used to improve memory of patients with senile dementia and cardiovascular diseases. However, little information on bioactive components from SSM or standardized extraction methods for these components is available. This study isolated and purified major components from SSM for the first time, and assessed their ability to inhibit soluble epoxide hydrolase (sEH). The results showed that acteoside is the most potent inhibitor of sEH, with an IC50 of 33.5 ± 0.5 µM. Additional active components, including harpagide, tryptophan, and 8-acetate-harpagide, along with acteoside, were tentatively identified using high-performance liquid chromatography photodiode array tandem mass spectrometry (HPLC-PDA-MS/MS) and quantified using an ultraviolet detector at 210 nm. Further, an ultrasonic-assisted extraction technique for extraction of four bioactive compounds in SSM was developed and optimized using response surface methodology (RSM). The optimal extraction conditions were: extraction time, 30.46 minutes; extraction temperature, 67.95 °C, and methanol concentration 53.85%. The prediction model of RSM was validated with laboratory experiments. The similarity between predicted and actual values was 97.84%. The extraction method is thus a rapid, environment-friendly, energy-saving method can be applied to extract bioactive components from SSM in large quantities.

Stachys sieboldii Miq. Root Attenuates Weight Gain and Dyslipidemia in Rats on a High-Fat and High-Cholesterol Diet.

This study aimed at investigating the anti-obesity and anti-dyslipidemic effects of Stachys sieboldii Miq. root (SS) powder in rats following a high-fat and high-cholesterol (HFC) diet for 6 weeks. Thirty-two Sprague-Dawley rats were fed one of the following diets: a regular diet (RD), HFC, HFC supplemented with 3% SS (HFC + 3SS) or HFC supplemented with 5% SS (HFC + 5SS). Following an HFC diet increased body weight (BW) gain (p < 0.001) and the food efficiency ratio (FER; p < 0.001); however, SS consumption gradually prevented the HFC-induced BW gain (p < 0.001) and increase in FER (p < 0.01). The HFC diet resulted in increased liver size (p < 0.001) and total adipose tissue weight (p < 0.001), whereas the SS supplementation decreased hepatomegaly (p < 0.05) and body fat mass (p < 0.001). SS consumption prevented the increased activities of serum alanine aminotransferase (ALT; p < 0.001), aspartate aminotransferase (AST; p < 0.001), alkaline phosphatase (ALP; p < 0.01 in HFC + 5SS) and lactate dehydrogenase (LDH; p < 0.001 in HFC + 5SS) induced by the HFC diet (p < 0.001). The SS supplementation improved lipid profiles in the circulation by lowering triglyceride (TG; p < 0.01), total cholesterol (TC; p < 0.001) and non-HDL cholesterol (non-HDL-C; p < 0.001) levels, as well as the atherogenic index (p < 0.01) and cardiac risk factor (p < 0.01). The lipid distribution in the liver (p < 0.05) and white adipose tissues (WAT; p < 0.001) of the HFC + SS diet-consuming rats was remarkably lower than that of the HFC diet-consuming rats. The average size of the epididymal adipose tissue (p < 0.001) was significantly lower in the HFC + SS diet-fed rats than in the HFC diet-fed rats. The fecal lipid (>3% SS; p < 0.001) and cholesterol (5% SS; p < 0.001) efflux levels were significantly elevated by the SS supplementation compared to those measured in the RD or HFC diet-fed groups. In addition, the hepatic lipid and cholesterol metabolism-related gene expressions were affected by SS consumption, as the hepatic anabolic gene expression (Acc; p < 0.001, Fas; p < 0.001 and G6pdh; p < 0.01) was significantly attenuated. The HFC + 5SS diet-fed rats exhibited elevated hepatic Cyp7a1 (p < 0.001), Hmgcr (p < 0.001) and Ldlr (p < 0.001) mRNA expression levels compared to the HFC diet-fed rats. These results suggest that SS may possess anti-adipogenic and lipid-lowering effects by enhancing lipid and cholesterol efflux in mammals.

New Rare Ent-Clerodane Diterpene Peroxides from Egyptian Mountain Tea (Qourtom) and Its Chemosystem as Herbal Remedies and Phytonutrients Agents.

Genus Stachys, the largest genera of the family Lamiaceae, and its species are frequently used as herbal teas due to their essential oils. Tubers of some Stachys species are also consumed as important nutrients for humans and animals due to their carbohydrate contents. Three new neo-clerodane diterpene peroxides, named stachaegyptin F-H (1, 2, and 4), together with two known compounds, stachysperoxide (3) and stachaegyptin A (5), were isolated from Stachys aegyptiaca aerial parts. Their structures were determined using a combination of spectroscopic techniques, including HR-FAB-MS and extensive 1D and 2D NMR (1H, 13C NMR, DEPT, 1H-1H COSY, HMQC, HMBC and NOESY) analyses. Additionally, a biosynthetic pathway for the isolated compounds (1-5) was discussed. The chemotaxonomic significance of the isolated diterpenoids of S. aegyptiaca in comparison to the previous reported ones from other Stachys species was also studied.

Genotoxic properties of Betonica officinalis, Gratiola officinalis, Vincetoxicum luteum and Vincetoxicum hirundinaria extracts.

Genotoxicity of B. officinalis, G. officinalis, V. luteum and V. hirundinaria extracts, which demonstrated strong antioxidant capacity, was tested using chromosome aberration, sister chromatid exchange (SCE), cytokinesis-block micronucleus and alkaline single-cell gel electrophoresis (comet) assays in human lymphocytes in vitro and Ames Salmonella/microsome test. All tested extracts were not mutagenic in S. typhimurium strains TA98 and TA100 with and without metabolic activation and did not induce chromosome aberrations in human lymphocytes in vitro. Extract from G. officinalis was the only one, which induced significant increase in micronuclei, indicating possible aneugenic effect. All investigated plant extracts induced DNA damage evaluated by the comet assay, while B. officinalis and V. luteum extracts induced slight increase in SCE values. The determined variation in response might be due to the plant extract tested and donor susceptibility.

Antiangiogenic Iridoids from Stachys ocymastrum and Premna resinosa.

In this paper, the isolation of one new iridoid glucoside, 6ß-acetoxyipolamiide (1: ), and thirteen (2:  - 14: ) known congeners from two Lamiaceae species, Stachys ocymastrum and Premna resinosa, leaf extracts is reported. The structural determination of the isolated compounds was performed by mono- and bidimensional NMR spectroscopic analysis as well as MS experiments. The isolates were assayed for their antiangiogenic activity by two in vivo models, zebrafish embryos and chick chorioallantoic membrane assays. The compounds with a significant antiangiogenic activity in both assays were ß-hydroxyipolamiide (2: ), ipolamiide (3: ), and buddlejoside A5 (8: ). 6-O-α-l-(3″-O-p-Methoxycinnamoyl-4″-O-acetyl)rhamnopyranosyl catalpol (13: ) and 6-O-α-l-(2″-trans-caffeoyl)rhamnopyranosyl catalpol (6: ) showed the best antiangiogenic response on blood vessel growth in zebrafish embryos, whereas saccatoside (10: ) and 6-O-α-l-(2″-O - : p-methoxycinnamoyl-3″-O-acetyl)rhamnopyranosyl catalpol (14: ) resulted in a strong reduction of capillary formation in the chorioallantoic membrane assay.

LC-ESI/LTQOrbitrap/MS/MS and GC-MS profiling of Stachys parviflora L. and evaluation of its biological activities.

The use of some Stachys genus as herbal remedies is known and the aerial parts have a pharmaceutical interest, being used in Anatolia and Iran as wild tea. In this study, chemical composition, antimicrobial, antioxidant, and antiproliferative activities of the methanolic extract and essential oil (EO) of Stachys parviflora L. (S. parviflora) were evaluated. Qualitative analysis of metabolites of S. parviflora methanolic extract was studied using liquid chromatography coupled to high resolution mass spectrometry (LC-ESI/LTQOrbitrap/MS), evidencing the presence of phenolic acids and flavonoids derivatives. The EO was analyzed using gas chromatography coupled to mass spectrometry (GC/MS). Eighty-seven compounds were characterized in the EO of S. parviflora, of which α-terpenyl acetate (23.6%), ß-caryophyllene (16.8%), bicyclogermacrene (9.3%), spathulenol (4.9%) and α-pinene (4.2%) were found to be the major components. The highest antimicrobial effect of EO was found to S. aureus and B. cereus (MIC = 0.01 µg/ml), while the highest activity of extract was against B. cereus (MIC = 125 µg/ml). The methanolic extract exhibited strong antioxidant activity in DPPH (IC50 = 76.87 µg/ml) and ß-carotene/linoleic acid assay (BCB, IC50 = 188.47 µg/ml) methods. Furthermore, in vitro cytotoxicity evaluation against three cell lines namely human ovarian carcinoma (A2780), human colon carcinoma (HCT), and mouse melanoma cell line (B16F10), showed an anti-proliferative activity of the EO ranging from IC50 value 30.95 µg/ml to 16.55 µg/ml. The results from this study have demonstrated the promising cytotoxic, antibacterial, and antifungal properties of S. parviflora, which could have wide potential applications in food and pharmaceutical industries.

Gas chromatography-mass spectrometry analysis and antimicrobial, antioxidant and anti-cancer activities of essential oils and extracts of Stachys schtschegleevii plant as biological macromolecules.

This work reports the chemical composition and antimicrobial activity of the essential oil as well as antioxidant and anti-cancer activities of ethyl acetate and aqueous extracts from leaf and stem of Stachys schtschegleevii. The structure of volatile parts and effective phenolic compounds as biological macromolecules as well as biological activity of the Stachys schtschegleevii plant were studied using various methods. Essential oils of stem and leaf were identified by steam distillation method and characterization was performed using a gas chromatography-mass spectrometry (GC-MS) analysis. Anti-microbial activity of extracted macromolecules from essential oils of two ethyl acetate and aqueous parts of stem and leaf were analyzed by minimal inhibition concentration (MIC) method. The crude extracts exhibited no appreciable antioxidant activities, in 2,2-diphenyl-1-picrylhydrazyl (DPPH) test by IC50 317.98 and 328.09 µg/mL for aqueous and ethyl acetate fractions, respectively. But, antioxidant activities for butylated hydroxytoluene (BHT) as a standard antioxidant showed significant results (18.16 µg/mL). Also, it was confirmed that antioxidant activities with the phenolic and flavonoid content of the plant for the aqueous and ethyl acetate elements were obtained 57.73, 59.81 and 21.95, 26.124 µg/mg, respectively. Furthermore, in ß-carotene/linoleic acid assay, inhibition percentages were recorded 71.17 and 96.24 for the aqueous and ethyl acetate elements, respectively. Therefore, ethyl acetate section of the plant showed antioxidant activity more than aqueous part. Hydrogen peroxide scavenging assay evaluated antioxidant activity for aqueous and the ethyl acetate extracts sections as 565 and 560 µg/mL, respectively. Also, power determinations of iron (III) oxide antioxidants from aqueous and ethyl acetate fractions were 6113 and 2.69, respectively. Furthermore, Anti-cancer activity of aqueous and ethyl acetate components evaluated in brine shrimp lethality test with LC50 about 100 and 700 µg/mL for aqueous and ethyl acetate fractions, respectively. The results showed that aqueous section species has a much lower effect than the calculated standard, but ethyl acetate section of the plant sample shows a more favorable effect than the standard. Therefore, the plant screened good antimicrobial, antioxidant and anti-cancer activities for its essential oil and ethyl acetate extract.

Antioxidant and cytoprotective effects of Stachys riederi var. japonica ethanol extract on UVA­irradiated human dermal fibroblasts.

Stachys riederi is one of the largest genera in the flowering plant family Lamiaceae. The aqueous extract of Stachys riederi var. japonica is known for its anti­allergic effect. In the present study, the antioxidant and cytoprotective effects of Stachys riederi var. japonica ethanol extract (SREE) on ultraviolet A (UVA)­irradiated human dermal fibroblasts (HDFs) were evaluated. At 100 µg/ml, SREE significantly inhibited production of reactive oxygen species (ROS) in UVA­irradiated HDFs. SREE at 100 µg/ml additionally markedly interfered with the loss of mitochondrial membrane potential (ΔΨm) in these cells. In addition, SREE at 100 µg/ml attenuated UVA­induced DNA fragmentation and caspase­3 activation in HDFs. SREE at 100 µg/ml additionally increased mRNA and protein expressions of Bcl­2 and decreased those of Bax and cytochrome c in UVA­irradiated HDFs. In summary, to the best of our knowledge, these results demonstrate for the first time that SREE exhibited antioxidant and cytoprotective effects on UVA­irradiated HDFs, which may be mediated through suppression of ROS generation, inhibition of the loss of ΔΨm and inhibition of apoptosis.

Ultrasound combined with manganese-oxide nanoparticles loaded on activated carbon for extraction and pre-concentration of thymol and carvacrol in methanolic extracts of Thymus daenensis, Salvia officinalis, Stachys pilifera, Satureja khuzistanica, and mentha, and water samples.

A dispersive micro solid-phase extraction (DMSPE) technique was developed using manganese-oxide nanoparticles loaded on activated carbon (Mn3O4-NPs-AC) as an effective sorbent combined with ultrasound for the extraction and determination of a trace amount of thymol and carvacrol in methanolic extracts of Thymus daenensis, Salvia officinalis, Stachys pilifera, Satureja khuzistanica and mentha, and water samples. Thymol and carvacrol phenolic compounds were extracted from real samples using acetonitrile (ACN) as the desorption solvent. Using central composite design (CCD), the effects of pH, ionic strength (NaCl), nano-sorbent mass, contact time, and desorption volume were investigated. Additionally, based on five-level variables, response surface methodology was used to determine the individual and interactive effects between factors on the process. The optimized extraction conditions included 12 mg of Mn3O4-NPs-AC as the sorbent, 300 µL of ACN as the desorption solvent, pH 3.0, 0.5 w/v% of NaCl, and 4.5 min sonication time. Under the optimized conditions, for all the samples, the limits of detection were 0.054-0.104 ng mL-1 and the limits of quantification were 0.178-0.345 ng mL-1. The correlation coefficients of the calibration curves were >0.985, i.e. in the range of 0.4-6000 ng mL-1. To validate the effects of the matrix, the recovery, reproducibility, repeatability, and overall uncertainty were calculated for the five methanolic extracts, at 50, 100, and 500 ng mL-1. The recovery ranged between 94.5% and 109.0% with a relative standard deviation of <8.0% for the repeatability and reproducibility precision, which strongly supports the favorable repeatability and reproducibility of the method. The presented method also has the excellent sorbent features of NPs for the sorption of the analyte, which is due to the use of ultrasound for dispersion of the material in the sample matrix.

A syringic acid derivative and two iridoid glycosides from the roots of Stachys geobombycis and their antioxidant properties.

A new derivative of syringic acid, stageobester A (1) and two iridoid glycosides which including a new one, stageoboside A (2), were isolated from the roots of Stachys geobombycis. Their structures were elucidated on the basis of spectroscopic methods, including extensive NMR and MS spectra. In addition, all the isolates were tested for their antioxidant capacity. Compounds 1 and 2 showed moderate antioxidant activities against DPPH radical, with IC50 values of 113.33 ± 1.53 and 40.33 ± 2.08 mg/L, respectively.